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32 results on '"MESH: Pituitary Adenylate Cyclase-Activating Polypeptide"'

1. Intranasal Administration of PACAP Is an Efficient Delivery Route to Reduce Infarct Volume and Promote Functional Recovery After Transient and Permanent Middle Cerebral Artery Occlusion

Author

Asma Cherait, Julie Maucotel, Benjamin Lefranc, Jérôme Leprince, David Vaudry, Lebon, Alexis, Différenciation et communication neuronale et neuroendocrine (DC2N), Université de Rouen Normandie (UNIROUEN), Normandie Université (NU)-Normandie Université (NU)-Institut National de la Santé et de la Recherche Médicale (INSERM), University of Algiers 3 : Université d' Alger 3, Plate-Forme de Recherche en Imagerie Cellulaire de Haute-Normandie (PRIMACEN), Normandie Université (NU)-Normandie Université (NU)-Institute for Research and Innovation in Biomedicine (IRIB), Normandie Université (NU)-Normandie Université (NU)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Centre National de la Recherche Scientifique (CNRS)-Université de Rouen Normandie (UNIROUEN), Normandie Université (NU)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Centre National de la Recherche Scientifique (CNRS)-High-tech Research Infrastructures for Life Sciences (HeRacLeS), and Normandie Université (NU)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Centre National de la Recherche Scientifique (CNRS)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Centre National de la Recherche Scientifique (CNRS)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Centre National de la Recherche Scientifique (CNRS)

Subjects
Male, 0301 basic medicine, Nasal cavity, functional recovery, Endocrinology, Diabetes and Metabolism, MESH: Neurotransmitter Agents, Pharmacology, infarct volume, lcsh:Diseases of the endocrine glands. Clinical endocrinology, cerebral ischemia, Mice, Endocrinology, 0302 clinical medicine, [CHIM] Chemical Sciences, MESH: Animals, MESH: Administration, Intranasal, Middle cerebral artery occlusion, Original Research, Neurotransmitter Agents, MESH: Infarction, Middle Cerebral Artery, Infarction, Middle Cerebral Artery, MESH: Neuroprotective Agents, MESH: Recovery of Function, pituitary adenylate cyclase-activating polypeptide, Neuroprotective Agents, medicine.anatomical_structure, Infarct volume, Brain Infarction, Central nervous system, Ischemia, Neuroprotection, intranasal administration, 03 medical and health sciences, MESH: Brain Infarction, MESH: Mice, Inbred C57BL, medicine, [CHIM]Chemical Sciences, Animals, MESH: Mice, Administration, Intranasal, lcsh:RC648-665, business.industry, MESH: Pituitary Adenylate Cyclase-Activating Polypeptide, Recovery of Function, medicine.disease, Functional recovery, MESH: Male, Mice, Inbred C57BL, 030104 developmental biology, Nasal administration, business, 030217 neurology & neurosurgery
Abstract

International audience; Intranasal (IN) administration appears to be a suitable route for clinical use as it allows direct delivery of bioactive molecules to the central nervous system, reducing systemic exposure and sides effects. Nevertheless, only some molecules can be transported to the brain from the nasal cavity. This led us to compare the efficiency of an IN, intravenous (IV), and intraperitoneal (IP) administration of pituitary adenylate cyclase-activating polypeptide (PACAP) after transient or permanent middle cerebral artery occlusion (MCAO) in C57BL/6 mice. The results show that the neuroprotective effect of PACAP is much more efficient after IN administration than IV injection while IP injection had no effect. IN administration of PACAP reduced the infarct volume when injected within 6 h after the reperfusion and improved functional recovery up to at least 1 week after the ischemia.

Published
2021
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2. Pituitary adenylate cyclase-activating polypeptide and vasoactive intestinal polypeptide promote the genesis of calcium currents in differentiating mouse embryonic stem cells

Author

Pierre Gressens, M. Chafai, William Rostène, Hubert Vaudry, M. Basille, Bruno J. Gonzalez, Estelle Louiset, Ludovic Galas, Différenciation et communication neuronale et neuroendocrine (DC2N), Université de Rouen Normandie (UNIROUEN), Normandie Université (NU)-Normandie Université (NU)-Institut National de la Santé et de la Recherche Médicale (INSERM), Imagerie cellulaire des neurorécepteurs et physiopathologie neuroendocrinienne, Université Pierre et Marie Curie - Paris 6 (UPMC)-Institut National de la Santé et de la Recherche Médicale (INSERM), Neuroprotection du Cerveau en Développement / Promoting Research Oriented Towards Early Cns Therapies (PROTECT), Assistance publique - Hôpitaux de Paris (AP-HP) (AP-HP)-Hôpital Robert Debré-Université Paris Diderot - Paris 7 (UPD7)-Institut National de la Santé et de la Recherche Médicale (INSERM), Neuroendocrinologie cellulaire et moléculaire, Normandie Université (NU), and Institut National de la Santé et de la Recherche Médicale (INSERM)

Subjects
MESH: Cell Differentiation, MESH: Neural Stem Cells, Neurogenesis, Cellular differentiation, Vasoactive intestinal peptide, Biology, Real-Time Polymerase Chain Reaction, Mice, MESH: Gene Expression Profiling, 03 medical and health sciences, 0302 clinical medicine, Neural Stem Cells, MESH: Reverse Transcriptase Polymerase Chain Reaction, MESH: Gene Expression Regulation, Developmental, [SDV.BC.IC]Life Sciences [q-bio]/Cellular Biology/Cell Behavior [q-bio.CB], [SDV.MHEP.PHY]Life Sciences [q-bio]/Human health and pathology/Tissues and Organs [q-bio.TO], Animals, MESH: Animals, L-type calcium channel, MESH: Embryonic Stem Cells, MESH: Mice, Embryonic Stem Cells, 030304 developmental biology, 0303 health sciences, Voltage-dependent calcium channel, Reverse Transcriptase Polymerase Chain Reaction, MESH: Real-Time Polymerase Chain Reaction, Gene Expression Profiling, General Neuroscience, Calcium channel, MESH: Pituitary Adenylate Cyclase-Activating Polypeptide, MESH: Vasoactive Intestinal Peptide, T-type calcium channel, Gene Expression Regulation, Developmental, Cell Differentiation, MESH: Immunohistochemistry, [SDV.MHEP.EM]Life Sciences [q-bio]/Human health and pathology/Endocrinology and metabolism, Immunohistochemistry, Embryonic stem cell, Molecular biology, Neural stem cell, MESH: Neurogenesis, 3. Good health, MESH: Calcium Channels, Pituitary Adenylate Cyclase-Activating Polypeptide, Calcium Channels, hormones, hormone substitutes, and hormone antagonists, 030217 neurology & neurosurgery, Vasoactive Intestinal Peptide
Abstract

International audience; Identification of novel molecules that can induce neuronal differentiation of embryonic stem (ES) cells is essential for deciphering the molecular mechanisms of early development and for exploring cell therapy approaches. Pituitary adenylate cyclase-activating polypeptide (PACAP) and vasoactive intestinal polypeptide (VIP) are known to be implicated early during ontogenesis in cell proliferation and neuronal differentiation. The aim of the present study was to determine the effects of VIP and PACAP on functional differentiation of ES cells. Quantitative-reverse transcription-polymerase chain reaction analysis showed an inversion of the expression pattern of PAC1 and VPAC1 receptors with time. ES cells expressed genes encoding extracellular signal-regulated kinase 1 and 2 and c-jun amino terminal kinase1. ES cells also expressed T-type α1I and α1G, L-type α1C and α1D, and N-type α1B calcium channel subunit mRNAs. Both peptides modified the shape of undifferentiated ES cells into bipolar cells expressing the neuronal marker neuron-specific enolase (NSE). Immunostaining indicated that PACAP intensified T-type α1I subunit immunoreactivity, whereas VIP increased L-types α1C and α1D, as well as N-type α1B subunit. Electrophysiological recording showed that VIP and PACAP enhanced transient calcium current. Moreover, VIP generated sustained calcium current. These findings demonstrate that PACAP and VIP induce morphological and functional differentiation of ES cells into a neuronal phenotype. Both peptides promote functional maturation of calcium channel subunits, suggesting that they can facilitate the genesis of cellular excitability.

Published
2011
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3. PACAP and a novel stable analog protect rat brain from ischemia: Insight into the mechanisms of action

Author

Omar Touzani, Alain Fournier, Steve Bourgault, David Vaudry, Hubert Vaudry, Agnieszka Dejda, Tommy Seaborn, Différenciation et communication neuronale et neuroendocrine (DC2N), Université de Rouen Normandie (UNIROUEN), Normandie Université (NU)-Normandie Université (NU)-Institut National de la Santé et de la Recherche Médicale (INSERM), Institut Fédératif de Recherches Multidisciplinaires sur les Peptides (IFRMP 23), Institut National de la Santé et de la Recherche Médicale (INSERM)-Centre de Lutte Contre le Cancer Henri Becquerel Normandie Rouen (CLCC Henri Becquerel)-Université de Rouen Normandie (UNIROUEN), Normandie Université (NU)-Normandie Université (NU)-Université Le Havre Normandie (ULH), Normandie Université (NU)-CHU Rouen, Normandie Université (NU)-Centre National de la Recherche Scientifique (CNRS), International Associated laboratory Samuel de Champlain, Institut National de la Recherche Scientifique [Québec] (INRS)-Université de Rouen Normandie (UNIROUEN), Institut Armand Frappier (INRS-IAF), Institut National de la Recherche Scientifique [Québec] (INRS)-Réseau International des Instituts Pasteur (RIIP), Hypoxie, physiopathologies cérébrovasculaire et tumorale (CERVOxy), Imagerie et Stratégies Thérapeutiques des pathologies Cérébrales et Tumorales (ISTCT), Université de Caen Normandie (UNICAEN), Normandie Université (NU)-Normandie Université (NU)-Centre National de la Recherche Scientifique (CNRS)-Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Université de Caen Normandie (UNICAEN), Normandie Université (NU)-Normandie Université (NU)-Centre National de la Recherche Scientifique (CNRS)-Commissariat à l'énergie atomique et aux énergies alternatives (CEA), Plate-Forme de Recherche en Imagerie Cellulaire de Haute-Normandie (PRIMACEN), Normandie Université (NU)-Normandie Université (NU)-Institute for Research and Innovation in Biomedicine (IRIB), Normandie Université (NU)-Normandie Université (NU)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Centre National de la Recherche Scientifique (CNRS)-Université de Rouen Normandie (UNIROUEN), Normandie Université (NU)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Centre National de la Recherche Scientifique (CNRS), This work was supported by INSERM, the European Institute for Peptide Research (IFRMP23), the Interreg 4A FEDER project TC2N, an ANR Jeune Chercheuse-Jeune Chercheur, the LARC Neurosciences network, and the Région Haute-Normandie, CHU Rouen, Normandie Université (NU)-Université de Rouen Normandie (UNIROUEN), Normandie Université (NU)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Centre de Lutte Contre le Cancer Henri Becquerel Normandie Rouen (CLCC Henri Becquerel)-Centre National de la Recherche Scientifique (CNRS), Normandie Université (NU)-Normandie Université (NU)-Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Centre National de la Recherche Scientifique (CNRS)-Université de Caen Normandie (UNICAEN), Normandie Université (NU)-Normandie Université (NU)-Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Centre National de la Recherche Scientifique (CNRS), Normandie Université (NU)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Centre National de la Recherche Scientifique (CNRS)-High-tech Research Infrastructures for Life Sciences (HeRacLeS), and Normandie Université (NU)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Centre National de la Recherche Scientifique (CNRS)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Centre National de la Recherche Scientifique (CNRS)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Centre National de la Recherche Scientifique (CNRS)

Subjects
Male, MESH: Inflammation, Physiology, Gene Expression, Apoptosis, Pharmacology, Biochemistry, MESH: Dose-Response Relationship, Drug, Brain ischemia, 0302 clinical medicine, Endocrinology, Drug Stability, Ischemia, MESH: Reverse Transcriptase Polymerase Chain Reaction, MESH: Caspase 3, MESH: Animals, bcl-2-Associated X Protein, 0303 health sciences, biology, Caspase 3, Reverse Transcriptase Polymerase Chain Reaction, Brain, MESH: Infarction, Middle Cerebral Artery, Nitric oxide synthase 2, Infarction, Middle Cerebral Artery, MESH: Neuroprotective Agents, Macrophage Inflammatory Proteins, 3. Good health, Stroke, Pituitary adenylate cyclase-activating peptide, Neuroprotective Agents, [SDV.TOX]Life Sciences [q-bio]/Toxicology, Pituitary Adenylate Cyclase-Activating Polypeptide, medicine.symptom, medicine.medical_specialty, Programmed cell death, MESH: Gene Expression, MESH: Rats, MESH: Macrophage Inflammatory Proteins, Neuroprotection, MESH: Stroke, MESH: Brain, 03 medical and health sciences, Cellular and Molecular Neuroscience, MESH: Drug Stability, Internal medicine, medicine, Animals, MESH: bcl-2-Associated X Protein, RNA, Messenger, Rats, Wistar, MESH: RNA, Messenger, 030304 developmental biology, Inflammation, Dose-Response Relationship, Drug, Interleukin-6, Tumor Necrosis Factor-alpha, MESH: Apoptosis, MESH: Pituitary Adenylate Cyclase-Activating Polypeptide, MESH: Rats, Wistar, medicine.disease, MESH: Interleukin-6, MESH: Male, Rats, Disease Models, Animal, Mechanism of action, MESH: Tumor Necrosis Factor-alpha, biology.protein, MESH: Ischemia, MESH: Disease Models, Animal, 030217 neurology & neurosurgery
Abstract

International audience; Pituitary adenylate cyclase-activating polypeptide (PACAP) shows potent protective effects in numerous models of neurological insults. However, the use of PACAP as a clinically efficient drug is limited by its poor metabolic stability. By combining identification of enzymatic cleavage sites with targeted chemical modifications, a metabolically stable and potent PACAP38 analog was recently developed. The neuroprotective activity of this novel compound was for the first time evaluated and compared to the native peptide using a rat model of middle cerebral artery occlusion (MCAO). Our results show that as low as picomolar doses of PACAP38 and its analog strongly reduce infarct volume and improve neurological impairment induced by stroke. In particular, these peptides inhibit the expression of Bcl-2-associated death promoter, caspase 3, macrophage inflammatory protein-1α, inducible nitric oxide synthase 2, tumor necrosis factor-α mRNAs, and increase extracellular signal-regulated kinase 2, B-cell CLL/lymphoma 2 and interleukin 6 mRNA levels. These results indicate that the neuroprotective effect of PACAP after MCAO is not only due to its ability to inhibit apoptosis but also to modulate the inflammatory response. The present study highlights the potential therapeutic efficacy of very low concentrations of PACAP or its metabolically stable derivative for the treatment of stroke.

Published
2011
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4. Pituitary adenylate cyclase-activating polypeptide inhibits caspase-3 activity but does not protect cerebellar granule neurons against β-amyloid (25–35)-induced apoptosis

Author

Alain Fournier, Hubert Vaudry, Magali Basille, Anthony Falluel-Morel, Bruno J. Gonzalez, David Vaudry, Cécile Cottet-Rousselle, Neuroendocrinologie cellulaire et moléculaire, Université de Rouen Normandie (UNIROUEN), Normandie Université (NU)-Normandie Université (NU)-Institut National de la Santé et de la Recherche Médicale (INSERM), Institut Armand Frappier (INRS-IAF), Institut National de la Recherche Scientifique [Québec] (INRS)-Réseau International des Instituts Pasteur (RIIP), and This work was supported by grants from INSERM (U413), an INSERM-FRSQ exchange program (to AF and HV) and the Conseil Regional de Haute-Normandie. A.F.M. was a recipient of a fellowship from the Conseil Régional de Haute-Normandie (LARC-Neuroscience network)

Subjects
β-amyloid peptide, Cerebellum, Physiology, Clinical Biochemistry, MESH: Neurons, Apoptosis, MESH: Neurotransmitter Agents, PACAP, MESH: Neuropeptides, Biochemistry, 0302 clinical medicine, Endocrinology, MESH: Caspase 3, Insulin, MESH: Animals, MESH: Peptide Fragments, Cells, Cultured, Caspase, Neurons, Neurotransmitter Agents, 0303 health sciences, biology, Caspase 3, Caspase Inhibitors, MESH: Amyloid beta-Peptides, Mitochondria, Cell biology, Pituitary adenylate cyclase-activating peptide, medicine.anatomical_structure, MESH: Cell Survival, Caspases, Enzyme Induction, [SDV.TOX]Life Sciences [q-bio]/Toxicology, Pituitary Adenylate Cyclase-Activating Polypeptide, MESH: Cells, Cultured, medicine.medical_specialty, MESH: Enzyme Induction, MESH: Rats, MESH: Caspase Inhibitors, Neurite, Cell Survival, MESH: Mitochondria, MESH: Insulin, Cysteine Proteinase Inhibitors, Neuroprotection, MESH: Cysteine Proteinase Inhibitors, 03 medical and health sciences, Cellular and Molecular Neuroscience, Internal medicine, medicine, Animals, Nerve Growth Factors, 030304 developmental biology, Amyloid beta-Peptides, MESH: Caspases, MESH: Nerve Growth Factors, MESH: Apoptosis, Neuropeptides, MESH: Pituitary Adenylate Cyclase-Activating Polypeptide, Neurotoxicity, Granule cell, medicine.disease, Peptide Fragments, MESH: Cerebellum, Rats, biology.protein, Granule cells, 030217 neurology & neurosurgery
Abstract

International audience; The beta-amyloid (Abeta) peptide Abeta25-35 provokes apoptosis of cerebellar granule cells through activation of caspase-3 while the neuropeptide pituitary adenylate cyclase-activating polypeptide (PACAP) promotes granule cell survival by inhibiting caspase-3 activation through the intrinsic apoptotic pathway. The aim of the present study was to determine whether PACAP could prevent Abeta25-35 neurotoxicity by inhibiting caspase-3 activity. A 24-h exposure of cultured cerebellar granule cells to Abeta25-35 induced shrinkage of cell bodies, neurite retraction and alteration of mitochondrial activity. Administration of graded concentrations (10-80 microM) of Abeta25-35 induced a dose-related decrease of the number of living cells, and the neurotoxic effect was highly significant after a 24-h exposure to 80 microM Abeta25-35. Exposure of cerebellar granule cells to Abeta25-35 markedly enhanced caspase-3 but not caspase-9 activity. Co-incubation with 1 microM PACAP significantly reduced Abeta25-35-evoked caspase-3 activation. In contrast, PACAP did not prevent the deleterious effects of Abeta25-35 on mitochondrial potential and granule cell survival. Taken together, these data suggest that caspase-3 activation is not the main pathway activated by Abeta25-35 that leads to granule cell death. The results also demonstrate that PACAP cannot be considered as a potent neuroprotective factor against Abeta25-35-induced apoptosis in cerebellar granule neurons.

Published
2004
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5. Delayed Pituitary Adenylate Cyclase–Activating Polypeptide Delivery After Brain Stroke Improves Functional Recovery by Inducing M2 Microglia/Macrophage Polarization

Author

Victor May, David Vaudry, Coralie Brifault, Marjorie Gras, Olivier Wurtz, Donovan Liot, Neuroendocrinologie cellulaire et moléculaire, Université de Rouen Normandie (UNIROUEN), Normandie Université (NU)-Normandie Université (NU)-Institut National de la Santé et de la Recherche Médicale (INSERM), University of Vermont College of Medicine, University of Vermont [Burlington], International Associated laboratory Samuel de Champlain, Institut National de la Recherche Scientifique [Québec] (INRS)-Université de Rouen Normandie (UNIROUEN), Différenciation et communication neuronale et neuroendocrine (DC2N), Institut National de la Santé et de la Recherche Médicale (INSERM)-Institut National de la Recherche Scientifique [Québec] (INRS)-Université de Rouen Normandie (UNIROUEN), and Normandie Université (NU)-Normandie Université (NU)

Subjects
Male, Time Factors, [SDV.NEU.NB]Life Sciences [q-bio]/Neurons and Cognition [q-bio.NC]/Neurobiology, microglia, Pharmacology, [SDV.IMM.II]Life Sciences [q-bio]/Immunology/Innate immunity, Mice, Drug Delivery Systems, MESH: Animals, Stroke, Microglia, Brain, Cell Polarity, MESH: Recovery of Function, stroke, pituitary adenylate cyclase-activating polypeptide, MESH: Microglia, medicine.anatomical_structure, [SDV.IMM.IA]Life Sciences [q-bio]/Immunology/Adaptive immunology, medicine.symptom, Stem cell, MESH: Stem Cell Transplantation, MESH: Cell Polarity, MESH: Injections, Intraventricular, Cardiology and Cardiovascular Medicine, Programmed cell death, Mice, 129 Strain, MESH: Mice, Transgenic, MESH: Drug Delivery Systems, Ischemia, Adenylate kinase, Inflammation, Mice, Transgenic, Neuroprotection, MESH: Stroke, MESH: Brain, MESH: Mice, 129 Strain, medicine, Animals, MESH: Mice, Injections, Intraventricular, Advanced and Specialized Nursing, business.industry, Macrophages, MESH: Pituitary Adenylate Cyclase-Activating Polypeptide, MESH: Time Factors, MESH: Macrophages, Recovery of Function, medicine.disease, MESH: Male, inflammation, Immunology, Neurology (clinical), business, Stem Cell Transplantation
Abstract

Background and Purpose— Until now, except thrombolysis, the therapeutical strategies targeting the acute phase of cerebral ischemia have been proven ineffective, and no approach is available to attenuate the delayed cell death mechanisms and the resulting functional deficits in the late phase. Then, we investigated whether a targeted and delayed delivery of pituitary adenylate cyclase–activating polypeptide (PACAP), a peptide known to exert neuroprotective activities, may dampen delayed pathophysiological processes improving functional recovery. Methods— Three days after permanent focal ischemia, PACAP-producing stem cells were transplanted intracerebro ventricularly in nonimmunosuppressed mice. At 7 and 14 days post ischemia, the effects of this stem cell–based targeted delivery of PACAP on functional recovery, volume lesions, and inflammatory processes were analyzed. Results— The delivery of PACAP in the vicinity of the infarct zone 3 days post stroke promotes fast, stable, and efficient functional recovery. This was correlated with a modulation of the postischemic inflammatory response. Transcriptomic and Ingenuity Pathway Analysis–based bioinformatic analyses identified several gene networks, functions, and key transcriptional factors, such as nuclear factor-κB, C/EBP-β, and Notch/RBP-J as PACAP’s potential targets. Such PACAP-dependent immunomodulation was further confirmed by morphometric and phenotypic analyses of microglial cells showing increased number of Arginase-1 + cells in mice treated with PACAP-expressing cells specifically, demonstrating the redirection of the microglial response toward a neuroprotective M2 phenotype. Conclusions— Our results demonstrated that immunomodulatory strategies capable of redirecting the microglial response toward a neuroprotective M2 phenotype in the late phase of brain ischemia could represent attractive options for stroke treatment in a new and unexploited therapeutical window.

Published
2015
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6. Pituitary Adenylate Cyclase-Activating Polypeptide Reverses Ammonium Metavanadate-Induced Airway Hyperresponsiveness in Rats

Author

Sonia Rouatbi, David Vaudry, Badreddine Sriha, Khémais Ben Rhouma, Olivier Wurtz, Mounira Tlili, Olfa Tebourbi, Mohsen Sakly, Laboratoire d'Informatique, de Parallélisme et de Productique (LIP2), Faculté des Sciences Mathématiques, Physiques et Naturelles de Tunis (FST), Université de Tunis El Manar (UTM)-Université de Tunis El Manar (UTM), Université de Sousse, Faculté des Sciences de Bizerte [Université de Carthage], Université de Carthage - University of Carthage, Physiologie Intégrée, Université de Carthage - University of Carthage-Université de Carthage - University of Carthage, International Associated laboratory Samuel de Champlain, Institut National de la Recherche Scientifique [Québec] (INRS)-Université de Rouen Normandie (UNIROUEN), Normandie Université (NU)-Normandie Université (NU)-Institut National de la Santé et de la Recherche Médicale (INSERM), Différenciation et communication neuronale et neuroendocrine (DC2N), and Université de Rouen Normandie (UNIROUEN)

Subjects
Male, Aging, Chemokine, MESH: Macrophages, Alveolar, [SDV.NEU.NB]Life Sciences [q-bio]/Neurons and Cognition [q-bio.NC]/Neurobiology, Chemokine CXCL2, Environmental pollution, medicine.disease_cause, Biochemistry, [SDV.IMM.II]Life Sciences [q-bio]/Immunology/Innate immunity, chemistry.chemical_compound, MESH: Aerosols, Interleukin-1alpha, MESH: Animals, Respiratory system, MESH: Interleukin-1alpha, Lung, Chemokine CCL3, MESH: Oxidative Stress, biology, Chemistry, lcsh:Cytology, MESH: Real-Time Polymerase Chain Reaction, General Medicine, MESH: Administration, Inhalation, 3. Good health, MESH: Respiratory Hypersensitivity, [SDV.IMM.IA]Life Sciences [q-bio]/Immunology/Adaptive immunology, Pituitary Adenylate Cyclase-Activating Polypeptide, Tumor necrosis factor alpha, Research Article, medicine.medical_specialty, Article Subject, MESH: Rats, Adenylate kinase, Real-Time Polymerase Chain Reaction, Proinflammatory cytokine, Ammonium metavanadate, Internal medicine, Administration, Inhalation, Macrophages, Alveolar, medicine, Respiratory Hypersensitivity, MESH: Chemokine CXCL2, Animals, MESH: Lung, MESH: Chemokine CCL3, lcsh:QH573-671, Rats, Wistar, Aerosols, Tumor Necrosis Factor-alpha, MESH: Pituitary Adenylate Cyclase-Activating Polypeptide, Cell Biology, MESH: Rats, Wistar, MESH: Male, Rats, Oxidative Stress, Endocrinology, MESH: Vanadates, 13. Climate action, MESH: Tumor Necrosis Factor-alpha, biology.protein, Vanadates, Oxidative stress
Abstract

The rate of atmospheric vanadium is constantly increasing due to fossil fuel combustion. This environmental pollution favours vanadium exposure in particular to its vanadate form, causing occupational bronchial asthma and bronchitis. Based on the well admitted bronchodilator properties of the pituitary adenylate cyclase-activating polypeptide (PACAP), we investigated the ability of this neuropeptide to reverse the vanadate-induced airway hyperresponsiveness in rats. Exposure to ammonium metavanadate aerosols (5 mg/m3/h) for 15 minutes induced 4 hours later an array of pathophysiological events, including increase of bronchial resistance and histological alterations, activation of proinflammatory alveolar macrophages, and increased oxidative stress status. Powerfully, PACAP inhalation (0.1 mM) for 10 minutes alleviated many of these deleterious effects as demonstrated by a decrease of bronchial resistance and histological restoration. PACAP reduced the level of expression of mRNA encoding inflammatory chemokines (MIP-1α, MIP-2, and KC) and cytokines (IL-1αand TNF-α) in alveolar macrophages and improved the antioxidant status. PACAP reverses the vanadate-induced airway hyperresponsiveness not only through its bronchodilator activity but also by counteracting the proinflammatory and prooxidative effects of the metal. Then, the development of stable analogs of PACAP could represent a promising therapeutic alternative for the treatment of inflammatory respiratory disorders.

Published
2014
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7. Biochemical Characterization of a Caspase-3 Far-red Fluorescent Probe for Non-invasive Optical Imaging of Neuronal Apoptosis

Author

Valérie Jolivel, Alain Fournier, Pierre-Yves Renard, Jérôme Leprince, Béatrice Botia, Heidi Ligeret, Omar Touzani, Hubert Vaudry, Bruno Delest, Anthony Romieu, David Vaudry, Marc Massonneau, Sébastien Arthaud, Pauline Noack, Christophe Portal, Guillaume Clavé, Différenciation et communication neuronale et neuroendocrine (DC2N), Université de Rouen Normandie (UNIROUEN), Normandie Université (NU)-Normandie Université (NU)-Institut National de la Santé et de la Recherche Médicale (INSERM), Institute for Research and Innovation in Biomedicine (IRIB), Normandie Université (NU)-Normandie Université (NU)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Centre National de la Recherche Scientifique (CNRS), International Associated laboratory Samuel de Champlain, Institut National de la Recherche Scientifique [Québec] (INRS)-Université de Rouen Normandie (UNIROUEN), QUantitative Imaging in Drug Development (QUIDD), QUantitative Imaging in Drug Development, Plate-Forme de Recherche en Imagerie Cellulaire de Haute-Normandie (PRIMACEN), Normandie Université (NU)-Normandie Université (NU)-Institute for Research and Innovation in Biomedicine (IRIB), Normandie Université (NU)-Normandie Université (NU)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Centre National de la Recherche Scientifique (CNRS)-Université de Rouen Normandie (UNIROUEN), Normandie Université (NU)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Centre National de la Recherche Scientifique (CNRS)-High-tech Research Infrastructures for Life Sciences (HeRacLeS), Normandie Université (NU)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Centre National de la Recherche Scientifique (CNRS)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Centre National de la Recherche Scientifique (CNRS)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Centre National de la Recherche Scientifique (CNRS), Chimie Organique et Bioorganique : Réactivité et Analyse (COBRA), Institut de Chimie Organique Fine (IRCOF), Normandie Université (NU)-Normandie Université (NU)-Institut national des sciences appliquées Rouen Normandie (INSA Rouen Normandie), Institut National des Sciences Appliquées (INSA)-Normandie Université (NU)-Institut National des Sciences Appliquées (INSA)-Centre National de la Recherche Scientifique (CNRS)-Université de Rouen Normandie (UNIROUEN), Institut National des Sciences Appliquées (INSA)-Normandie Université (NU)-Institut National des Sciences Appliquées (INSA)-Centre National de la Recherche Scientifique (CNRS)-Institut Normand de Chimie Moléculaire Médicinale et Macromoléculaire (INC3M), Université de Caen Normandie (UNICAEN), Normandie Université (NU)-Normandie Université (NU)-École Nationale Supérieure d'Ingénieurs de Caen (ENSICAEN), Normandie Université (NU)-Université Le Havre Normandie (ULH), Normandie Université (NU)-Université de Rouen Normandie (UNIROUEN), Normandie Université (NU)-Institut national des sciences appliquées Rouen Normandie (INSA Rouen Normandie), Institut National des Sciences Appliquées (INSA)-Normandie Université (NU)-Institut National des Sciences Appliquées (INSA)-Institut de Chimie du CNRS (INC)-Centre National de la Recherche Scientifique (CNRS)-Université de Caen Normandie (UNICAEN), Normandie Université (NU)-École Nationale Supérieure d'Ingénieurs de Caen (ENSICAEN), Normandie Université (NU)-Institut de Chimie du CNRS (INC)-Centre National de la Recherche Scientifique (CNRS)-Centre National de la Recherche Scientifique (CNRS), Hypoxie, physiopathologies cérébrovasculaire et tumorale (CERVOxy), Imagerie et Stratégies Thérapeutiques des pathologies Cérébrales et Tumorales (ISTCT), Normandie Université (NU)-Normandie Université (NU)-Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Centre National de la Recherche Scientifique (CNRS)-Université de Caen Normandie (UNICAEN), Normandie Université (NU)-Normandie Université (NU)-Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Centre National de la Recherche Scientifique (CNRS), Institut Armand Frappier (INRS-IAF), Institut National de la Recherche Scientifique [Québec] (INRS)-Réseau International des Instituts Pasteur (RIIP), Normandie Université (NU)-Normandie Université (NU)-Centre National de la Recherche Scientifique (CNRS)-Institut National de la Santé et de la Recherche Médicale (INSERM), Normandie Université (NU)-Normandie Université (NU)-Centre National de la Recherche Scientifique (CNRS)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Université de Rouen Normandie (UNIROUEN), Normandie Université (NU)-Centre National de la Recherche Scientifique (CNRS)-Institut National de la Santé et de la Recherche Médicale (INSERM), Institut Normand de Chimie Moléculaire Médicinale et Macromoléculaire (INC3M), Institut de Chimie du CNRS (INC)-École Nationale Supérieure d'Ingénieurs de Caen (ENSICAEN), Institut National des Sciences Appliquées (INSA)-Normandie Université (NU)-Institut National des Sciences Appliquées (INSA)-Université Le Havre Normandie (ULH), Normandie Université (NU)-Centre National de la Recherche Scientifique (CNRS)-Université de Caen Normandie (UNICAEN), Normandie Université (NU)-Institut de Chimie du CNRS (INC)-École Nationale Supérieure d'Ingénieurs de Caen (ENSICAEN), Normandie Université (NU)-Centre National de la Recherche Scientifique (CNRS)-Institut de Chimie Organique Fine (IRCOF), and Institut National des Sciences Appliquées (INSA)-Normandie Université (NU)-Institut National des Sciences Appliquées (INSA)-Centre National de la Recherche Scientifique (CNRS)-Centre National de la Recherche Scientifique (CNRS)

Subjects
medicine.medical_treatment, MESH: Neurons, Apoptosis, Proteomics, Imaging, Mice, [SDV.MHEP.MI]Life Sciences [q-bio]/Human health and pathology/Infectious diseases, MESH: Caspase 3, MESH: Animals, Caspase, Cells, Cultured, Neurons, biology, Caspase 3, Optical Imaging, Brain, General Medicine, Carbocyanines, MESH: Fluorescent Dyes, 3. Good health, Cell biology, Stroke, Caspase-3, [SDV.TOX]Life Sciences [q-bio]/Toxicology, MESH: Oligopeptides, Pituitary Adenylate Cyclase-Activating Polypeptide, [SDV.NEU]Life Sciences [q-bio]/Neurons and Cognition [q-bio.NC], MESH: Carbocyanines, Oligopeptides, MESH: Cells, Cultured, Programmed cell death, MESH: Rats, MESH: Rhodamines, Cellular and Molecular Neuroscience, MESH: Brain, In vivo, MESH: Mice, Inbred C57BL, medicine, Animals, [CHIM]Chemical Sciences, Rats, Wistar, MESH: Mice, Fluorescent Dyes, MESH: Optical Imaging, Protease, Rhodamines, MESH: Apoptosis, MESH: Pituitary Adenylate Cyclase-Activating Polypeptide, MESH: Rats, Wistar, Molecular biology, In vitro, Rats, Mice, Inbred C57BL, biology.protein, [SDV.SP.PHARMA]Life Sciences [q-bio]/Pharmaceutical sciences/Pharmacology
Abstract

International audience; Apoptosis is a regulated process, leading to cell death, which is involved in several pathologies including neurodegenerative diseases and stroke. Caspase-3 is a key enzyme of the apoptotic pathway and is considered as a major target for the treatment of abnormal cell death. Sensitive and non-invasive methods to monitor caspase-3 activity in cells and in the brain of living animals are needed to test the efficiency of novel therapeutic strategies. In the present study, we have biochemically characterized a caspase-3 far-red fluorescent probe, QCASP3.2, that can be used to detect apoptosis in vivo. The specificity of cleavage of QCASP3.2 was demonstrated using recombinant caspases and protease inhibitors. The functionality of the probe was also established in cere-bellar neurons cultured in apoptotic conditions. QCASP3.2 did not exhibit any toxicity and appeared to accurately reflect the induction and inhibition of caspase activity by H 2 O 2 and PACAP, respectively, both in cell lysates and in cultured neurons. Finally, intravenous injection of the probe after cere-bral ischemia revealed activation of caspase-3 in the infarcted hemisphere. Thus, the present study demonstrates that QCASP3.2 is a suitable probe to monitor apoptosis both in vitro and in vivo and illustrates some of the possible applications of this caspase-3 fluorescent probe.

Published
2014
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8. Induction of serpinb1a by PACAP or NGF is required for PC12 cells survival after serum withdrawal

Author

Aurélia Ravni, Tommy Seaborn, Olivier Wurtz, Ruby Au, Hubert Vaudry, Lee E. Eiden, David Vaudry, Alain Fournier, Billy K. C. Chow, Différenciation et communication neuronale et neuroendocrine (DC2N), Université de Rouen Normandie (UNIROUEN), Normandie Université (NU)-Normandie Université (NU)-Institut National de la Santé et de la Recherche Médicale (INSERM), International Associated laboratory Samuel de Champlain, Institut National de la Recherche Scientifique [Québec] (INRS)-Université de Rouen Normandie (UNIROUEN), Plate-Forme de Recherche en Imagerie Cellulaire de Haute-Normandie (PRIMACEN), Normandie Université (NU)-Normandie Université (NU)-Institute for Research and Innovation in Biomedicine (IRIB), Normandie Université (NU)-Normandie Université (NU)-Centre National de la Recherche Scientifique (CNRS)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Université de Rouen Normandie (UNIROUEN), Normandie Université (NU)-Centre National de la Recherche Scientifique (CNRS)-Institut National de la Santé et de la Recherche Médicale (INSERM), Institut Armand Frappier (INRS-IAF), Réseau International des Instituts Pasteur (RIIP)-Institut National de la Recherche Scientifique [Québec] (INRS), National Institutes of Health [Bethesda] (NIH), This study was supported by INSERM (U982), the National Institute of Mental Health (NIMH) Intramural Research Program, an INSERM-FRSQ program (to A. F. and D. V.), the Interreg TC2N project, and the Région Haute-Normandie., Laflamme, Nancy, Normandie Université (NU)-Normandie Université (NU)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Centre National de la Recherche Scientifique (CNRS)-Université de Rouen Normandie (UNIROUEN), Normandie Université (NU)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Centre National de la Recherche Scientifique (CNRS)-High-tech Research Infrastructures for Life Sciences (HeRacLeS), Normandie Université (NU)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Centre National de la Recherche Scientifique (CNRS)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Centre National de la Recherche Scientifique (CNRS)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Centre National de la Recherche Scientifique (CNRS), Institut National de la Recherche Scientifique [Québec] (INRS)-Réseau International des Instituts Pasteur (RIIP), Institut Fédératif de Recherches Multidisciplinaires sur les Peptides (IFRMP 23), CHU Rouen, Normandie Université (NU)-Normandie Université (NU)-Université Le Havre Normandie (ULH), Normandie Université (NU)-Université de Rouen Normandie (UNIROUEN), Normandie Université (NU)-CRLCC Henri Becquerel-Institut National de la Santé et de la Recherche Médicale (INSERM)-Centre National de la Recherche Scientifique (CNRS), Imager Lab, Columbia University [New York], Neuroendocrinologie cellulaire et moléculaire, Différenciation et communication neuronale et neuroendocrine ( DC2N ), Université de Rouen Normandie ( UNIROUEN ), Normandie Université ( NU ) -Normandie Université ( NU ) -Institut National de la Santé et de la Recherche Médicale ( INSERM ), Institut National de la Santé et de la Recherche Médicale ( INSERM ) -Institut National de la Recherche Scientifique [Québec] ( INRS ) -Université de Rouen Normandie ( UNIROUEN ), Normandie Université ( NU ) -Normandie Université ( NU ), Department of Pediatrics, Hôpital St-François d'Assise, Centre de Recherche du Centre Hospitalier Universitaire de Québec ( CRCHUQ ), Plate-Forme de Recherche en Imagerie Cellulaire de Haute-Normandie ( PRIMACEN ), Normandie Université ( NU ) -Normandie Université ( NU ) -Institute for Research and Innovation in Biomedicine ( IRIB ), Normandie Université ( NU ) -Normandie Université ( NU ) -Institut National de la Santé et de la Recherche Médicale ( INSERM ) -Université de Rouen Normandie ( UNIROUEN ), Normandie Université ( NU ) -Institut National de la Santé et de la Recherche Médicale ( INSERM ), Department de zoology, The University of Hong Kong ( HKU ), Institut Armand Frappier ( INRS-IAF ), Institut National de la Recherche Scientifique [Québec] ( INRS ) -Réseau International des Instituts Pasteur ( RIIP ) -Institut Armand Frappier, Laboratory of Cellular and Molecular Regulation, National Institute of Mental Health, and This study was supported by INSERM (U982), the National Institute of Mental Health (NIMH) Intramural Research Program, an INSERM-FRSQprogram (to A. F. and D. V.), the Interreg TC2N project, and the Région Haute-Normandie.

Subjects
[SDV.NEU.NB]Life Sciences [q-bio]/Neurons and Cognition [q-bio.NC]/Neurobiology, Tropomyosin receptor kinase A, Biochemistry, [SDV.IMM.II]Life Sciences [q-bio]/Immunology/Innate immunity, PC12 Cells, Culture Media, Serum-Free, [CHIM] Chemical Sciences, MESH: Animals, Caspase, MESH: Nerve Growth Factor, biology, Kinase, MESH: Culture Media, Serum-Free, pituitary adenylate cyclase-activating polypeptide, 3. Good health, [SDV.TOX] Life Sciences [q-bio]/Toxicology, cell death, [SDV.IMM.IA]Life Sciences [q-bio]/Immunology/Adaptive immunology, MESH: Cell Survival, [SDV.SP.PHARMA] Life Sciences [q-bio]/Pharmaceutical sciences/Pharmacology, [SDV.TOX]Life Sciences [q-bio]/Toxicology, neuroprotection, hormones, hormone substitutes, and hormone antagonists, Neurotrophin, medicine.medical_specialty, Programmed cell death, endocrine system, MESH: Rats, p38 mitogen-activated protein kinases, caspase, cell survival, Article, nerve growth factor, Cellular and Molecular Neuroscience, Internal medicine, MESH: Serpins, medicine, Animals, MESH: PC12 Cells, [CHIM]Chemical Sciences, Protein kinase A, Serpins, [ SDV ] Life Sciences [q-bio], [ SDV.BC ] Life Sciences [q-bio]/Cellular Biology, MESH: Pituitary Adenylate Cyclase-Activating Polypeptide, [ SDV.BC.BC ] Life Sciences [q-bio]/Cellular Biology/Subcellular Processes [q-bio.SC], Rats, Nerve growth factor, Endocrinology, nervous system, biology.protein, [SDV.SP.PHARMA]Life Sciences [q-bio]/Pharmaceutical sciences/Pharmacology
Abstract

Thumbnail image of graphical abstract PC12 cells are used to study the signaling mechanisms underlying the neurotrophic and neuroprotective activities of pituitary adenylate cyclase-activating polypeptide (PACAP) and nerve growth factor (NGF). Previous microarray experiments indicated that serpinb1a was the most induced gene after 6 h of treatment with PACAP or NGF. This study confirmed that serpinb1a is strongly activated by PACAP and NGF in a time-dependent manner with a maximum induction (~ 50-fold over control) observed after 6 h of treatment. Co-incubation with PACAP and NGF resulted in a synergistic up-regulation of serpinb1a expression (200-fold over control), suggesting that PACAP and NGF act through complementary mechanisms. Consistently, PACAP-induced serpinb1a expression was not blocked by TrkA receptor inhibition. Nevertheless, the stimulation of serpinb1a expression by PACAP and NGF was significantly reduced in the presence of extracellular signal-regulated kinase, calcineurin, protein kinase A, p38, and PI3K inhibitors, indicating that the two trophic factors share some common pathways in the regulation of serpinb1a. Finally, functional investigations conducted with siRNA revealed that serpinb1a is not involved in the effects of PACAP and NGF on PC12 cell neuritogenesis, proliferation or body cell volume but mediates their ability to block caspases 3/7 activity and to promote PC12 cell survival. Pituitary adenylate cyclase-activating polypeptide (PACAP) and nerve growth factor (NGF) induce a strong increase in serpinb1 a expression in PC12 cells. Functional investigations revealed that this increase in serpinb1a does not affect cell proliferation or differentiation but inhibits caspase 3 activity and promotes cell survival., link_to_OA_fulltext

Published
2014
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9. Cortical-layer-specific effects of PACAP and tPA on interneuron migration during post-natal development of the cerebellum

Author

Emilie Raoult, Magalie Bénard, Hitoshi Komuro, Denis Vivien, Hubert Vaudry, Alain Fournier, David Vaudry, Alexis Lebon, Ludovic Galas, Laflamme, Nancy, Plate-Forme de Recherche en Imagerie Cellulaire de Haute-Normandie (PRIMACEN), Université de Rouen Normandie (UNIROUEN), Normandie Université (NU)-Normandie Université (NU)-Institute for Research and Innovation in Biomedicine (IRIB), Normandie Université (NU)-Normandie Université (NU)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Centre National de la Recherche Scientifique (CNRS)-Université de Rouen Normandie (UNIROUEN), Normandie Université (NU)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Centre National de la Recherche Scientifique (CNRS)-High-tech Research Infrastructures for Life Sciences (HeRacLeS), Normandie Université (NU)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Centre National de la Recherche Scientifique (CNRS)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Centre National de la Recherche Scientifique (CNRS)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Centre National de la Recherche Scientifique (CNRS), Différenciation et communication neuronale et neuroendocrine (DC2N), Normandie Université (NU)-Normandie Université (NU)-Institut National de la Santé et de la Recherche Médicale (INSERM), Institute for Research and Innovation in Biomedicine (IRIB), Normandie Université (NU)-Normandie Université (NU)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Centre National de la Recherche Scientifique (CNRS), International Associated laboratory Samuel de Champlain, Institut National de la Recherche Scientifique [Québec] (INRS)-Université de Rouen Normandie (UNIROUEN), Cleveland Clinic, Sérine protéases et physiopathologie de l'unité neurovasculaire, Université de Caen Normandie (UNICAEN), Institut Armand Frappier (INRS-IAF), Institut National de la Recherche Scientifique [Québec] (INRS)-Réseau International des Instituts Pasteur (RIIP), This work was supported by the Institute for Research and Innovation in Biomedicine (IRIB), the Cell Imaging Platform of Normandy (PRIMACEN), INSERM, the FEDER (# 2517), Interreg 4A TC2N European project, the LARC-Neurosciences Network, and the Region Haute-Normandie., Normandie Université (NU)-Normandie Université (NU)-Centre National de la Recherche Scientifique (CNRS)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Université de Rouen Normandie (UNIROUEN), Normandie Université (NU)-Centre National de la Recherche Scientifique (CNRS)-Institut National de la Santé et de la Recherche Médicale (INSERM), Normandie Université (NU)-Normandie Université (NU)-Centre National de la Recherche Scientifique (CNRS)-Institut National de la Santé et de la Recherche Médicale (INSERM), and Réseau International des Instituts Pasteur (RIIP)-Institut National de la Recherche Scientifique [Québec] (INRS)

Subjects
Male, Cerebellum, PACAP, Biochemistry, Tissue plasminogen activator, MESH: Plasminogen Activator Inhibitor 1, Interneuron migration, chemistry.chemical_compound, 0302 clinical medicine, Cell Movement, MESH: Tissue Plasminogen Activator, [CHIM] Chemical Sciences, MESH: Plasminogen, MESH: Animals, MESH: Cell Movement, 0303 health sciences, interneuron migration, Immunohistochemistry, MESH: Interneurons, Cell biology, [SDV.TOX] Life Sciences [q-bio]/Toxicology, medicine.anatomical_structure, Tissue Plasminogen Activator, Plasminogen activator inhibitor-1, [SDV.SP.PHARMA] Life Sciences [q-bio]/Pharmaceutical sciences/Pharmacology, [SDV.TOX]Life Sciences [q-bio]/Toxicology, Pituitary Adenylate Cyclase-Activating Polypeptide, basket and stellate cells, Female, medicine.drug, Interneuron, Internal granular layer, cerebellum, MESH: Rats, MESH: Cytoplasmic Granules, Cytoplasmic Granules, Cerebellar Cortex, 03 medical and health sciences, Cellular and Molecular Neuroscience, Organ Culture Techniques, Interneurons, Plasminogen Activator Inhibitor 1, medicine, Animals, [CHIM]Chemical Sciences, tPA, Rats, Wistar, 030304 developmental biology, MESH: Cerebellar Cortex, MESH: Pituitary Adenylate Cyclase-Activating Polypeptide, granule neurons, Plasminogen, MESH: Immunohistochemistry, MESH: Rats, Wistar, MESH: Cerebellum, MESH: Male, MESH: Organ Culture Techniques, Rats, chemistry, nervous system, Hepatic stellate cell, [SDV.SP.PHARMA]Life Sciences [q-bio]/Pharmaceutical sciences/Pharmacology, Neuroscience, Plasminogen activator, MESH: Female, 030217 neurology & neurosurgery
Abstract

International audience; During early post-natal development of the cerebellum, granule neurons (GN) execute a centripetal migration toward the internal granular layer, whereas basket and stellate cells (B/SC) migrate centrifugally to reach their final position in the molecular layer (ML). We have previously shown that pituitary adenylate cyclase-activating polypeptide (PACAP) stimulates in vitro the expression and release of the serine protease tissue-type plasminogen activator (tPA) from GN, but the coordinated role of PACAP and tPA during interneuron migration has not yet been investigated. Here, we show that endogenous PACAP is responsible for the transient arrest phase of GN at the level of the Purkinje cell layer (PCL) but has no effect on B/SC. tPA is devoid of direct effect on GN motility in vitro, although it is widely distributed along interneuron migratory routes in the ML, PCL, and internal granular layer. Interestingly, plasminogen activator inhibitor 1 reduces the migration speed of GN in the ML and PCL, and that of B/SC in the ML. Taken together, these results reveal for the first time that tPA facilitates the migration of both GN and fast B/SC at the level of their intersection in the ML through degradation of the extracellular matrix. Crucial role of tissue plasminogen activator (tPA) in interneuron migration. Interneuron migration is a critical step for normal establishment of neuronal network. This study indicates that, in the post-natal cerebellum, tPA facilitates the opposite migration of immature excitatory granule neurons (GN) and immature inhibitory basket/stellate cells (B/SC) along the same migratory route. These data show that tPA exerts a pivotal role in neurodevelopment.

Published
2014
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10. Design and characterization of novel cell-penetrating peptides from pituitary adenylate cyclase-activating polypeptide

Author

Hubert Vaudry, David Vaudry, Ngoc-Duc Doan, Myriam Létourneau, Nicolas Doucet, David Chatenet, Benjamin Folch, Alain Fournier, Institut Armand Frappier (INRS-IAF), Réseau International des Instituts Pasteur (RIIP)-Institut National de la Recherche Scientifique [Québec] (INRS), International Associated laboratory Samuel de Champlain, Institut National de la Recherche Scientifique [Québec] (INRS)-Université de Rouen Normandie (UNIROUEN), Normandie Université (NU)-Normandie Université (NU)-Institut National de la Santé et de la Recherche Médicale (INSERM), Différenciation et communication neuronale et neuroendocrine (DC2N), Université de Rouen Normandie (UNIROUEN), and This work was supported by the CIHR grant #102734 (Alain Fournier) and NSERC Discovery grant GPIN 402623-2011 (Nicolas Doucet). Nicolas Doucet also acknowledges supports from the FRQNT Strategic Cluster 'Regroupement Québécois de Recherche sur la Fonction, la Structure et l'Ingénierie des Protéines' (PROTEO) and the FRQS Strategic Cluster 'Groupe de Recherche Axé sur la Structure des Protéines' (GRASP).

Subjects
media_common.quotation_subject, Green Fluorescent Proteins, Endogenous cell-penetrating peptide, Pharmaceutical Science, MESH: Cricetinae, CHO Cells, Cell-Penetrating Peptides, MESH: Drug Design, Endocytosis, PACAP, Transfection, 03 medical and health sciences, 0302 clinical medicine, MESH: Green Fluorescent Proteins, Cricetulus, MESH: Cricetulus, MESH: CHO Cells, MESH: Plasmids, Cellular uptake, Cricetinae, Animals, Humans, MESH: Animals, Internalization, 030304 developmental biology, media_common, MESH: Cell-Penetrating Peptides, 0303 health sciences, MESH: Humans, Chemistry, Pinocytosis, MESH: Transfection, HEK 293 cells, MESH: Pituitary Adenylate Cyclase-Activating Polypeptide, In vitro, HEK293 Cells, Biochemistry, Lipofectamine, MESH: HEK293 Cells, [SDV.TOX]Life Sciences [q-bio]/Toxicology, Drug Design, Drug delivery, Pituitary Adenylate Cyclase-Activating Polypeptide, hormones, hormone substitutes, and hormone antagonists, 030217 neurology & neurosurgery, Intracellular, Plasmids
Abstract

International audience; The discovery of cell-penetrating peptide opened up new promising avenues for the non-invasive delivery of non-permeable biomolecules within the intracellular compartment. However, some setbacks such as possible toxic effects or unexpected immunological responses have limited their use in clinic. To overcome these obstacles, we investigated the use of novel cell-penetrating peptides (CPPs) derived from the endogenous neuropeptide Pituitary adenylate cyclase-activating polypeptide (PACAP). First, we demonstrated the propensity of native PACAP isoforms (PACAP27 and PACAP38) to efficiently deliver a large and non-permeable molecule, i.e. streptavidin, into cells. An inactive modified fragment of PACAP38, i.e. [Arg(17)]PACAP(11-38), with preserved cell-penetrating physico-chemical properties, was also synthesized and successfully use for the intracellular delivery of various cargoes such as small molecules, peptides, proteins, and polynucleotides. Especially, its effectiveness as a transfection agent was comparable to Lipofectamine 2000 while being non-toxic for cells. Uptake mechanism studies demonstrated that direct translocation, caveolae-dependent endocytosis and macropinocytosis were involved in the internalization of [Arg(17)]PACAP(11-38). This study not only opened up a new aspect in the usefulness of PACAP and its derivatives for therapeutic application but also contributed to the identification of new members of the CPP family. As such, inactive PACAP-related analogs could represent excellent vectors for in vitro and in vivo applications.

Published
2012
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11. Receptor-independent cellular uptake of pituitary adenylate cyclase-activating polypeptide

Author

Myriam Létourneau, Ngoc-Duc Doan, Hubert Vaudry, Alain Fournier, David Chatenet, David Vaudry, Institut Armand Frappier (INRS-IAF), Réseau International des Instituts Pasteur (RIIP)-Institut National de la Recherche Scientifique [Québec] (INRS), Université Laval [Québec] (ULaval), Différenciation et communication neuronale et neuroendocrine (DC2N), Université de Rouen Normandie (UNIROUEN), Normandie Université (NU)-Normandie Université (NU)-Institut National de la Santé et de la Recherche Médicale (INSERM), and Financial support was obtained from the CIHR

Subjects
Male, Intracrine, Transcription, Genetic, Receptors, Pituitary Adenylate Cyclase-Activating Polypeptide, MESH: Amino Acid Sequence, MESH: Rats, Sprague-Dawley, MESH: Protein Isoforms, Mass Spectrometry, Rats, Sprague-Dawley, 0302 clinical medicine, Cytosol, MESH: Cytosol, Testis, Protein Isoforms, MESH: Animals, Receptor, 0303 health sciences, MESH: Testis, Pinocytosis, Intracellular receptor, Endocytosis, Biochemistry, MESH: Cell Survival, MESH: Calcium, [SDV.TOX]Life Sciences [q-bio]/Toxicology, Intranuclear calcium release, MESH: Endocytosis, Pituitary Adenylate Cyclase-Activating Polypeptide, Intracellular, hormones, hormone substitutes, and hormone antagonists, Fluorescein-5-isothiocyanate, Protein Binding, MESH: Cell Nucleus, endocrine system, MESH: Rats, Cell Survival, Molecular Sequence Data, Adenylate kinase, Translocation, Biology, Cell Line, 03 medical and health sciences, MESH: Fluorescein-5-isothiocyanate, MESH: Protein Binding, Animals, Humans, Amino Acid Sequence, Molecular Biology, 030304 developmental biology, MESH: Mass Spectrometry, Cell Nucleus, MESH: Receptors, Pituitary Adenylate Cyclase-Activating Polypeptide, MESH: Humans, MESH: Molecular Sequence Data, Binding Sites, MESH: Transcription, Genetic, MESH: Pituitary Adenylate Cyclase-Activating Polypeptide, Cell Membrane, Cell Biology, MESH: Male, MESH: Cell Line, Rats, Nuclear receptor, MESH: Binding Sites, Intracrine factor, Calcium, 030217 neurology & neurosurgery, MESH: Cell Membrane
Abstract

International audience; Pituitary adenylate cyclase-activating polypeptide (PACAP), a hypophysiotropic neurohormone, participates in the regulation of pleiotropic functions. The recent discovery of intracellular PACAP receptors in the brain and the testis as well as the physico-chemical characteristics of PACAP, i.e. extended α-helix containing basic residues, prompted us to evaluate the propensity of PACAP to cross the plasma membrane in a receptor-independent manner. Using confocal microscopy and flow cytometry, we demonstrated the ability of FITC-conjugated PACAP to efficiently penetrate into the internal cell compartment by direct translocation and endocytosis through clathrin-coated pits and macropinocytosis. Our study also revealed that, once inside the cells, PACAP38 is not entirely degraded by intracellular enzymes and that a significant amount of intact PACAP38 is also able to exit cells. Moreover, using binding assay on rat nuclear fractions from various tissues, PACAP nuclear receptors were identified. We also found that PACAP stimulates calcium release in rat testis nuclei. Interestingly, PACAP27 and PACAP38 but not VIP were able to upregulate de novo DNA synthesis in testis nuclei and that this effect was abolished by PACAP(6-38). These results support the presence of PAC1 receptors at the nuclear membrane and raise questions about their role in the biological activity of the peptide. These findings contribute to the characterization of PACAP as an intracrine factor and suggest that these intracellular PAC1 binding sites, probably associated with specific biological activities, should be taken into account during the development of PACAP-based drugs.

Published
2011
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12. The PACAP-regulated gene selenoprotein T is highly induced in nervous, endocrine, and metabolic tissues during ontogenetic and regenerative processes

Author

Destiny-Love Manecka, Jean Lesage, Youssef Anouar, Salah Elias, Loubna Boukhzar, Abdeslam Chagraoui, Sébastien Arthaud, Isabelle Lihrmann, Gaëtan Prévost, Isabelle Dorval-Coiffec, Bernard Jégou, Yannick Tanguy, Didier Vieau, Anthony Falluel-Morel, Différenciation et communication neuronale et neuroendocrine (DC2N), Université de Rouen Normandie (UNIROUEN), Normandie Université (NU)-Normandie Université (NU)-Institut National de la Santé et de la Recherche Médicale (INSERM), Groupe d'Etude de la Reproduction Chez l'Homme et les Mammiferes (GERHM), Institut National de la Santé et de la Recherche Médicale (INSERM)-Université de Rennes 1 (UR1), Université de Rennes (UNIV-RENNES)-Université de Rennes (UNIV-RENNES), Unité Neurosciences et Physiologie Adaptative, Université de Lille, Sciences et Technologies, INSERM (U982), University of Rouen, the Conseil Régional de Haute-Normandie, Université de Rennes (UR)-Institut National de la Santé et de la Recherche Médicale (INSERM), and Brébion, Alice

Subjects
Male, Pituitary gland, Cerebellum, Thyroid Gland, PC12 Cells, MESH: Selenoproteins, 0302 clinical medicine, Endocrinology, MESH: Pituitary Gland, Testis, MESH: Animals, Selenoproteins, chemistry.chemical_classification, [SDV.MHEP.EM] Life Sciences [q-bio]/Human health and pathology/Endocrinology and metabolism, 0303 health sciences, MESH: Testis, Thyroid, MESH: Regeneration, Brain, Selenoprotein T, [SDV.MHEP.EM]Life Sciences [q-bio]/Human health and pathology/Endocrinology and metabolism, 3. Good health, medicine.anatomical_structure, Liver, Pituitary Gland, Pituitary Adenylate Cyclase-Activating Polypeptide, medicine.medical_specialty, MESH: Rats, Neuropeptide, Biology, 03 medical and health sciences, MESH: Brain, Internal medicine, medicine, Endocrine system, Animals, Regeneration, MESH: PC12 Cells, Progenitor cell, Rats, Wistar, 030304 developmental biology, MESH: Pituitary Adenylate Cyclase-Activating Polypeptide, MESH: Rats, Wistar, MESH: Male, Rats, MESH: Thyroid Gland, chemistry, Selenoprotein, 030217 neurology & neurosurgery, MESH: Liver
Abstract

International audience; Selenoproteins contain the essential trace element selenium whose deficiency leads to major disorders including cancer, male reproductive system failure, or autoimmune thyroid disease. Up to now, 25 selenoprotein-encoding genes were identified in mammals, but the spatiotemporal distribution, regulation, and function of some of these selenium-containing proteins remain poorly documented. Here, we found that selenoprotein T (SelT), a new thioredoxin-like protein, is regulated by the trophic neuropeptide pituitary adenylate cyclase-activating polypeptide (PACAP) in differentiating but not mature adrenomedullary cells. In fact, our analysis revealed that, in rat, SelT is highly expressed in most embryonic structures, and then its levels decreased progressively as these organs develop, to vanish in most adult tissues. In the brain, SelT was abundantly expressed in neural progenitors in various regions such as the cortex and cerebellum but was undetectable in adult nervous cells except rostral migratory-stream astrocytes and Bergmann cells. In contrast, SelT expression was maintained in several adult endocrine tissues such as pituitary, thyroid, or testis. In the pituitary gland, SelT was found in secretory cells of the anterior lobe, whereas in the testis, the selenoprotein was present only in spermatogenic and Leydig cells. Finally, we found that SelT expression is strongly stimulated in liver cells during the regenerative process that occurs after partial hepatectomy. Taken together, these data show that SelT induction is associated with ontogenesis, tissue maturation, and regenerative mechanisms, indicating that this PACAP-regulated selenoprotein may play a crucial role in cell growth and activity in nervous, endocrine, and metabolic tissues.

Published
2011
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13. Pituitary adenylate cyclase-activating polypeptide (PACAP) stimulates the expression and the release of tissue plasminogen activator (tPA) in neuronal cells: involvement of tPA in the neuroprotective effect of PACAP

Author

Raoult, Emilie, Roussel, Benoît Denis, Bénard, Magalie, Lefebvre, Thomas, Ravni, Aurélia, Ali, Carine, Vivien, Denis, Komuro, Hitoshi, Fournier, Alain, Vaudry, Hubert, Vaudry, David, Galas, Ludovic, Institut Fédératif de Recherches Multidisciplinaires sur les Peptides (IFRMP 23), Institut National de la Santé et de la Recherche Médicale (INSERM)-Centre de Lutte Contre le Cancer Henri Becquerel Normandie Rouen (CLCC Henri Becquerel)-Université de Rouen Normandie (UNIROUEN), Normandie Université (NU)-Normandie Université (NU)-Université Le Havre Normandie (ULH), Normandie Université (NU)-CHU Rouen, Normandie Université (NU)-Centre National de la Recherche Scientifique (CNRS), Différenciation et communication neuronale et neuroendocrine (DC2N), Université de Rouen Normandie (UNIROUEN), Normandie Université (NU)-Normandie Université (NU)-Institut National de la Santé et de la Recherche Médicale (INSERM), Plate-Forme de Recherche en Imagerie Cellulaire de Haute-Normandie (PRIMACEN), Normandie Université (NU)-Normandie Université (NU)-Institute for Research and Innovation in Biomedicine (IRIB), Normandie Université (NU)-Normandie Université (NU)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Centre National de la Recherche Scientifique (CNRS)-Université de Rouen Normandie (UNIROUEN), Normandie Université (NU)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Centre National de la Recherche Scientifique (CNRS), International Associated laboratory Samuel de Champlain, Institut National de la Recherche Scientifique [Québec] (INRS)-Université de Rouen Normandie (UNIROUEN), Sérine protéases et physiopathologie de l'unité neurovasculaire, Université de Caen Normandie (UNICAEN), Department of Neuroscience/NC30, Cleveland Clinic-Lerner Research Institute, Institut Armand Frappier (INRS-IAF), Institut National de la Recherche Scientifique [Québec] (INRS)-Réseau International des Instituts Pasteur (RIIP), and This work was supported by INSERM, the European Institute for Peptide Research (IFRMP23), the Cell Imaging Platform of Normandy (PRIMACEN), the FEDER program # 2517, the Interreg 4A TC2N project, the LARC-Neurosciences Network and the Région Haute-Normandie

Subjects
Male, endocrine system, MESH: Rats, Cell Survival, MESH: Neurons, Real-Time Polymerase Chain Reaction, PC12 Cells, Gene Expression Regulation, Enzymologic, Cell Movement, MESH: Tissue Plasminogen Activator, Cerebellum, MESH: PC12 Cells, Animals, MESH: Animals, cerebellar granule neurons, Enzyme Inhibitors, Rats, Wistar, MESH: Cell Movement, Neurons, integumentary system, MESH: Culture Media, Conditioned, MESH: Real-Time Polymerase Chain Reaction, MESH: Gene Expression Regulation, Enzymologic, MESH: Pituitary Adenylate Cyclase-Activating Polypeptide, MESH: Neuroprotective Agents, MESH: Rats, Wistar, MESH: Cerebellum, MESH: Male, Rats, Neuroprotective Agents, MESH: Cell Survival, MESH: Enzyme Inhibitors, [SDV.TOX]Life Sciences [q-bio]/Toxicology, Culture Media, Conditioned, Tissue Plasminogen Activator, Pituitary Adenylate Cyclase-Activating Polypeptide, neuroprotection, Female, MESH: Female, hormones, hormone substitutes, and hormone antagonists
Abstract

International audience; Pituitary adenylate cyclase-activating polypeptide (PACAP) and tissue plasminogen activator (tPA) play important roles in neuronal migration and survival. However, a direct link between the neurotrophic effects of PACAP and tPA has never been investigated. In this study, we show that, in PC12 cells, PACAP induced a 9.85-fold increase in tPA gene expression through activation of the protein kinase A- and protein kinase C-dependent signaling pathways. In immature cerebellar granule neurons (CGN), PACAP stimulated tPA mRNA expression and release of proteolytically active tPA. Immunocytochemical labeling revealed the presence of tPA in the cytoplasm and processes of cultured CGN. The inhibitory effect of PACAP on CGN motility was not affected by the tPA substrate plasminogen or the tPA inhibitor plasminogen activator inhibitor-1. In contrast, plasminogen activator inhibitor-1 significantly reduced the stimulatory effect of PACAP on CGN survival. Altogether, these data indicate that tPA gene expression is activated by PACAP in both tumoral and normal neuronal cells. The present study also demonstrates that PACAP stimulates the release of tPA which promotes CGN survival by a mechanism dependent of its proteolytic activity.

Published
2011
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14. Pituitary adenylate cyclase-activating polypeptide protects astroglial cells against oxidative stress-induced apoptosis

Author

Masmoudi-Kouki, Olfa, Douiri, Salma, Hamdi, Yosra, Kaddour, Hadhemi, Bahdoudi, Saima, Vaudry, David, Basille, Magali, Leprince, Jérôme, Fournier, Alain, Vaudry, Hubert, Tonon, Marie-Christine, Amri, Mohamed, Université de Tunis El Manar (UTM), Différenciation et communication neuronale et neuroendocrine (DC2N), Université de Rouen Normandie (UNIROUEN), Normandie Université (NU)-Normandie Université (NU)-Institut National de la Santé et de la Recherche Médicale (INSERM), Institut Fédératif de Recherches Multidisciplinaires sur les Peptides (IFRMP 23), Institut National de la Santé et de la Recherche Médicale (INSERM)-Centre de Lutte Contre le Cancer Henri Becquerel Normandie Rouen (CLCC Henri Becquerel)-Université de Rouen Normandie (UNIROUEN), Normandie Université (NU)-Normandie Université (NU)-Université Le Havre Normandie (ULH), Normandie Université (NU)-CHU Rouen, Normandie Université (NU)-Centre National de la Recherche Scientifique (CNRS), Plate-Forme de Recherche en Imagerie Cellulaire de Haute-Normandie (PRIMACEN), Normandie Université (NU)-Normandie Université (NU)-Institute for Research and Innovation in Biomedicine (IRIB), Normandie Université (NU)-Normandie Université (NU)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Centre National de la Recherche Scientifique (CNRS)-Université de Rouen Normandie (UNIROUEN), Normandie Université (NU)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Centre National de la Recherche Scientifique (CNRS), Institut Armand Frappier (INRS-IAF), and Institut National de la Recherche Scientifique [Québec] (INRS)-Réseau International des Instituts Pasteur (RIIP)

Subjects
MESH: Signal Transduction, endocrine system, MESH: Rats, MESH: Mitochondria, MESH: Drug Interactions, [SDV.NEU.NB]Life Sciences [q-bio]/Neurons and Cognition [q-bio.NC]/Neurobiology, MESH: Neurons, Apoptosis, [CHIM.THER]Chemical Sciences/Medicinal Chemistry, MESH: Animals, Newborn, Cerebellum, MESH: Caspase 3, [SDV.BC.IC]Life Sciences [q-bio]/Cellular Biology/Cell Behavior [q-bio.CB], Animals, Drug Interactions, MESH: Animals, Rats, Wistar, Cells, Cultured, Cerebral Cortex, Neurons, MESH: Glutathione, MESH: Oxidative Stress, MESH: Culture Media, Conditioned, Caspase 3, MESH: Apoptosis, MESH: Pituitary Adenylate Cyclase-Activating Polypeptide, MESH: Reactive Oxygen Species, Hydrogen Peroxide, MESH: Rats, Wistar, Glutathione, MESH: Cerebellum, MESH: Cerebral Cortex, Mitochondria, Rats, MESH: Astrocytes, Oxidative Stress, Animals, Newborn, Astrocytes, Culture Media, Conditioned, [SDV.SP.PHARMA]Life Sciences [q-bio]/Pharmaceutical sciences/Pharmacology, Pituitary Adenylate Cyclase-Activating Polypeptide, MESH: Hydrogen Peroxide, Reactive Oxygen Species, hormones, hormone substitutes, and hormone antagonists, Signal Transduction, MESH: Cells, Cultured
Abstract

International audience; Oxidative stress, associated with a variety of disorders including neurodegenerative diseases, results from accumulation of reactive oxygen species (ROS). Oxidative stress is not only responsible for neuron apoptosis, but can also provoke astroglial cell death. Numerous studies indicate that pituitary adenylate cyclase-activating polypeptide (PACAP) promotes neuron survival, but nothing is known regarding the action of PACAP on astroglial cell survival. Thus, the purpose of the present study was to investigate the potential glioprotective effect of PACAP on H(2)O(2)-induced astrocyte death. Pre-treatment of cultured rat astrocytes with nanomolar concentrations of PACAP prevented cell death provoked by H(2)O(2) (300 μM), whereas vasoactive intestinal polypeptide was devoid of protective activity. The effect of PACAP on astroglial cell survival was abolished by the type 1 PACAP receptor antagonist, PACAP6-38. The protective action of PACAP was blocked by the protein kinase A inhibitor H89, the protein kinase C inhibitor chelerythrine and the mitogen-activated protein (MAP)-kinase kinase (MEK) inhibitor U0126. PACAP stimulated glutathione formation, and blocked H(2)O(2)-evoked ROS accumulation and glutathione content reduction. In addition, PACAP prevented the decrease of mitochondrial activity and caspase 3 activation induced by H(2)O(2). Taken together, these data indicate for the first time that PACAP, acting through type 1 PACAP receptor, exerts a potent protective effect against oxidative stress-induced astrocyte death. The anti-apoptotic activity of PACAP on astrocytes is mediated through the protein kinase A, protein kinase C and MAPK transduction pathways, and can be accounted for by inhibition of ROS-induced mitochondrial dysfunctions and caspase 3 activation.

Published
2011
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15. Neuroprotective effects of PACAP against ethanol-induced toxicity in the developing rat cerebellum

Author

Vadim Le Joncour, Alain Fournier, Magalie Bénard, Vincent Roy, Mickaël Naassila, Hubert Vaudry, Valérie Jolivel, Béatrice Botia, David Vaudry, Delphine Burel, Différenciation et communication neuronale et neuroendocrine ( DC2N ), Université de Rouen Normandie ( UNIROUEN ), Normandie Université ( NU ) -Normandie Université ( NU ) -Institut National de la Santé et de la Recherche Médicale ( INSERM ), Institut Fédératif de Recherches Multidisciplinaires sur les Peptides ( IFRMP 23 ), CHU Rouen-Université Le Havre Normandie ( ULH ), Normandie Université ( NU ) -Normandie Université ( NU ) -Université de Rouen Normandie ( UNIROUEN ), Normandie Université ( NU ) -CRLCC Henri Becquerel-Institut National de la Santé et de la Recherche Médicale ( INSERM ) -Centre National de la Recherche Scientifique ( CNRS ), International Associated laboratory Samuel de Champlain, Institut National de la Recherche Scientifique [Québec] ( INRS ) -Université de Rouen Normandie ( UNIROUEN ), Neuroendocrinologie cellulaire et moléculaire, Psychologie et Neurosciences de la Cognition et de l'affectivité ( PSY-NCA ), Normandie Université ( NU ) -Normandie Université ( NU ), Alcoolisation précoce et vulnerabilité à la dépendance, Université de Picardie Jules Verne ( UPJV ) -Institut National de la Santé et de la Recherche Médicale ( INSERM ), Institut Armand Frappier ( INRS-IAF ), Institut National de la Recherche Scientifique [Québec] ( INRS ) -Réseau International des Instituts Pasteur ( RIIP ) -Institut Armand Frappier, Naassila, Mickael, Différenciation et communication neuronale et neuroendocrine (DC2N), Université de Rouen Normandie (UNIROUEN), Normandie Université (NU)-Normandie Université (NU)-Institut National de la Santé et de la Recherche Médicale (INSERM), Institut Fédératif de Recherches Multidisciplinaires sur les Peptides (IFRMP 23), CHU Rouen, Normandie Université (NU)-Normandie Université (NU)-Université Le Havre Normandie (ULH), Normandie Université (NU)-Université de Rouen Normandie (UNIROUEN), Normandie Université (NU)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Centre de Lutte Contre le Cancer Henri Becquerel Normandie Rouen (CLCC Henri Becquerel)-Centre National de la Recherche Scientifique (CNRS), Institut National de la Recherche Scientifique [Québec] (INRS)-Université de Rouen Normandie (UNIROUEN), Psychologie et Neurosciences de la Cognition et de l'affectivité (PSY-NCA), Normandie Université (NU)-Normandie Université (NU), Groupe de Recherche sur l'alcool et les pharmacodépendances - UMR INSERM_S 1247 (GRAP), Université de Picardie Jules Verne (UPJV)-Institut National de la Santé et de la Recherche Médicale (INSERM), Institut Armand Frappier (INRS-IAF), Institut National de la Recherche Scientifique [Québec] (INRS)-Réseau International des Instituts Pasteur (RIIP), Institut National de la Santé et de la Recherche Médicale (INSERM)-Centre de Lutte Contre le Cancer Henri Becquerel Normandie Rouen (CLCC Henri Becquerel)-Université de Rouen Normandie (UNIROUEN), Normandie Université (NU)-CHU Rouen, and Normandie Université (NU)-Centre National de la Recherche Scientifique (CNRS)

Subjects
Cerebellum, MESH : Cerebellum, Cellular differentiation, Toxicology, MESH : Ethanol, MESH: Animals, Newborn, 0302 clinical medicine, MESH : Neuroprotective Agents, MESH: Animals, 0303 health sciences, MESH : Rats, General Neuroscience, MESH : Animals, Newborn, Neurogenesis, MESH: Neuroprotective Agents, MESH: Motor Activity, MESH : Pituitary Adenylate Cyclase-Activating Polypeptide, medicine.anatomical_structure, Neuroprotective Agents, Toxicity, Pituitary Adenylate Cyclase-Activating Polypeptide, [SDV.NEU]Life Sciences [q-bio]/Neurons and Cognition [q-bio.NC], hormones, hormone substitutes, and hormone antagonists, medicine.medical_specialty, Programmed cell death, endocrine system, MESH: Ethanol, MESH: Rats, Neuropeptide, Biology, Motor Activity, MESH : Rats, Wistar, Neuroprotection, 03 medical and health sciences, Internal medicine, medicine, Animals, [SDV.NEU] Life Sciences [q-bio]/Neurons and Cognition [q-bio.NC], Rats, Wistar, 030304 developmental biology, Ethanol, MESH: Pituitary Adenylate Cyclase-Activating Polypeptide, MESH: Rats, Wistar, Granule cell, MESH: Cerebellum, Rats, Endocrinology, Animals, Newborn, [ SDV.NEU ] Life Sciences [q-bio]/Neurons and Cognition [q-bio.NC], MESH : Animals, MESH : Motor Activity, 030217 neurology & neurosurgery
Abstract

International audience; The developing rat cerebellum is particularly sensitive to alcohol at the end of the first postnatal week, a period of intense neurogenesis. The neuropeptide Pituitary adenylate cyclase-activating polypeptide (PACAP) has previously been shown to prevent the death of cultured neurons in vitro. We have thus investigated the capacity of PACAP to counteract ethanol toxicity in 8-day-old rats. Behavioral studies revealed that PACAP reduces the deleterious action of alcohol in the negative geotaxis test. Administration of ethanol induced a transient increase of the expression of pro-apoptotic genes including c-jun or caspase-3 , which could be partially blocked by PACAP. Alcohol inhibited the expression of the α6 GABA ( A ) subunit while PACAP increased neuroD2 mRNA level, two markers of neuronal differentiation. Although gene regulations occurred rapidly, a third injection of ethanol was required to strongly reduce the number of granule cells in the internal granule cell layer, an effect which was totally blocked by PACAP. The action of PACAP was mimicked by D-JNKi1 and Z-VAD-FMK, indicating the involvement of the jun and caspase-3 pathways in alcohol toxicity. The present data demonstrate that PACAP can counteract in vivo the deleterious effect of ethanol. The beneficial action of PACAP on locomotor activity precedes its activity on cell survival, indicating that PACAP can block the detrimental action of ethanol on cell differentiation.

Published
2011
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16. Design and in vitro characterization of PAC1/VPAC1-selective agonists with potent neuroprotective effects

Author

Michel Detheux, Agnieszka Dejda, David Vaudry, David Chatenet, Alain Fournier, Steve Bourgault, Hubert Vaudry, Myriam Létourneau, Ngoc-Duc Doan, Institut Armand Frappier (INRS-IAF), Institut National de la Recherche Scientifique [Québec] (INRS)-Réseau International des Instituts Pasteur (RIIP), International Associated laboratory Samuel de Champlain, Institut National de la Recherche Scientifique [Québec] (INRS)-Université de Rouen Normandie (UNIROUEN), Normandie Université (NU)-Normandie Université (NU)-Institut National de la Santé et de la Recherche Médicale (INSERM), Euroscreen S.A., Institut Fédératif de Recherches Multidisciplinaires sur les Peptides (IFRMP 23), Institut National de la Santé et de la Recherche Médicale (INSERM)-Centre de Lutte Contre le Cancer Henri Becquerel Normandie Rouen (CLCC Henri Becquerel)-Université de Rouen Normandie (UNIROUEN), Normandie Université (NU)-Normandie Université (NU)-Université Le Havre Normandie (ULH), Normandie Université (NU)-CHU Rouen, Normandie Université (NU)-Centre National de la Recherche Scientifique (CNRS), and This work was supported by the Canadian Institutes of Health Research

Subjects
Agonist, medicine.drug_class, Receptors, Vasoactive Intestinal Polypeptide, Type I, MESH: Neurons, MESH: Cricetinae, CHO Cells, MESH: Drug Design, Pharmacology, Biology, Transfection, Biochemistry, Neuroprotection, Calcium in biology, 03 medical and health sciences, Structure-Activity Relationship, MESH: Structure-Activity Relationship, 0302 clinical medicine, Cricetulus, MESH: Cricetulus, MESH: CHO Cells, Cricetinae, medicine, Animals, Humans, MESH: Animals, Receptor, 030304 developmental biology, Neurons, 0303 health sciences, MESH: Humans, MESH: Transfection, MESH: Pituitary Adenylate Cyclase-Activating Polypeptide, Dopaminergic, Rational design, MESH: Receptors, Vasoactive Intestinal Polypeptide, Type I, MESH: Neuroprotective Agents, In vitro, 3. Good health, Pituitary adenylate cyclase-activating peptide, Neuroprotective Agents, [SDV.TOX]Life Sciences [q-bio]/Toxicology, Drug Design, MESH: Receptors, Pituitary Adenylate Cyclase-Activating Polypeptide, Type I, Pituitary Adenylate Cyclase-Activating Polypeptide, 030217 neurology & neurosurgery, Receptors, Pituitary Adenylate Cyclase-Activating Polypeptide, Type I
Abstract

International audience; Pituitary adenylate cyclase-activating polypeptide (PACAP) is a pleiotropic neuropeptide that exerts a large array of actions in the central nervous system and periphery. Through the activation of PAC1 and VPAC1, PACAP is able to exert neuroprotective, as well as anti-inflammatory effects, two phenomena involved in the pathogenesis and the progression of neurodegenerative diseases. The aim of the current study was to provide insights into the molecular arrangement of the amino terminus of PACAP and to develop new potent and selective PAC1/VPAC1 agonists promoting neuronal survival. We have synthesized a series of PACAP derivatives and measured their binding affinity and their ability to induce intracellular calcium mobilization for each receptor, i.e. PAC1, VPAC1, and VPAC2. Ultimately, analogs with an improved pharmacological profile were evaluated in an in vitro model of neuronal loss. Results showed that introduction of a hydroxyproline or an alanine moiety, respectively, at position 2 or 7 generated derivatives without significant VPAC2 agonistic activity. Moreover, the structure-activity relationship study suggests the presence of common (Asx-turn like) and distinct (different N-capping type) secondary structures that might be responsible for receptor recognition, selectivity and activation. Finally, evaluation of the neuroprotective activity of [Ala(7)]PACAP27 and [Hyp(2)]PACAP27 demonstrated their ability to protect potently human dopaminergic SH-SY5Y neuroblasts against the toxicity of MPP(+), in pre- and co-treatment experiments. These new pharmacological and structural data should prove useful for the rational design of PACAP-derived compounds that could be putative therapeutic agents for the treatment of neurodegenerative diseases.

Published
2010
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17. Balanced effect of PACAP and FasL on granule cell death during cerebellar development: a morphological, functional and behavioural characterization

Author

Emma R. Isaac, Bruno J. Gonzalez, Delphine Burel, Ludovic Galas, Alain Fournier, Pierre Chapillon, Nancy M. Sherwood, Bénédicte Parent, Vincent Roy, Sébastien Arthaud, Hubert Vaudry, Arnaud Desfeux, Aurélie Allais, Institut Fédératif de Recherches Multidisciplinaires sur les Peptides (IFRMP 23), Institut National de la Santé et de la Recherche Médicale (INSERM)-Centre de Lutte Contre le Cancer Henri Becquerel Normandie Rouen (CLCC Henri Becquerel)-Université de Rouen Normandie (UNIROUEN), Normandie Université (NU)-Normandie Université (NU)-Université Le Havre Normandie (ULH), Normandie Université (NU)-CHU Rouen, Normandie Université (NU)-Centre National de la Recherche Scientifique (CNRS), Institute for Research and Innovation in Biomedicine (IRIB), Université de Rouen Normandie (UNIROUEN), Normandie Université (NU)-Normandie Université (NU)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Centre National de la Recherche Scientifique (CNRS), Department of Biology, University of Victoria [Canada] (UVIC), Endothélium microcirculatoire cérébral et lésions du système nerveux central au cours du développement (Néovasc), Normandie Université (NU)-Normandie Université (NU)-Institute for Research and Innovation in Biomedicine (IRIB), Normandie Université (NU)-Normandie Université (NU)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Centre National de la Recherche Scientifique (CNRS)-Université de Rouen Normandie (UNIROUEN), Normandie Université (NU)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Centre National de la Recherche Scientifique (CNRS)-Institut National de la Santé et de la Recherche Médicale (INSERM), Institut Armand Frappier (INRS-IAF), and Institut National de la Recherche Scientifique [Québec] (INRS)-Réseau International des Instituts Pasteur (RIIP)

Subjects
MESH: Cell Death, Cerebellum, Time Factors, MESH: Neurons, Biochemistry, MESH: Mice, Knockout, Fas ligand, MESH: Animals, Newborn, Mice, 0302 clinical medicine, MESH: Gene Expression Regulation, Developmental, MESH: Behavior, Animal, MESH: Caspase 3, MESH: Animals, Caspase, MESH: Cell Size, MESH: Bromodeoxyuridine, Mice, Knockout, Neurons, 0303 health sciences, MESH: Muscle, Skeletal, MESH: Statistics, Nonparametric, biology, Behavior, Animal, Cell Death, Caspase 3, Gene Expression Regulation, Developmental, MESH: Motor Activity, Pituitary adenylate cyclase-activating peptide, medicine.anatomical_structure, [SDV.TOX]Life Sciences [q-bio]/Toxicology, Pituitary Adenylate Cyclase-Activating Polypeptide, hormones, hormone substitutes, and hormone antagonists, medicine.medical_specialty, Programmed cell death, endocrine system, Fas Ligand Protein, MESH: Reflex, MESH: Psychomotor Performance, In Vitro Techniques, Motor Activity, Statistics, Nonparametric, 03 medical and health sciences, Cellular and Molecular Neuroscience, MESH: Mice, Inbred C57BL, Internal medicine, MESH: Cell Proliferation, Reflex, medicine, Animals, RNA, Messenger, Muscle, Skeletal, MESH: Mice, 030304 developmental biology, MESH: RNA, Messenger, Cell Proliferation, Cell Size, MESH: Pituitary Adenylate Cyclase-Activating Polypeptide, MESH: Time Factors, Granule cell, MESH: Cerebellum, MESH: Fas Ligand Protein, Mice, Inbred C57BL, Endocrinology, Animals, Newborn, Bromodeoxyuridine, Apoptosis, biology.protein, 030217 neurology & neurosurgery, Psychomotor Performance
Abstract

International audience; It is now established that the development of the CNS requires equilibrium between cell survival and apoptosis. Pituitary adenylate cyclase-activating polypeptide (PACAP) exerts a powerful protective effect on cerebellar granule cells by inhibiting the caspase 3. In contrast, Fas ligand (FasL) plays an essential role during ontogenesis in eliminating supernumerary neurons by apoptosis. To determine if PACAP and FasL interact during cerebellar development, we characterized the effects of these factors on cerebellar morphogenesis and caspase 3 activity in PACAP+/+ and PACAP-/- mice. First, we demonstrated in vivo that PACAP is able to reverse the diminution of internal granule cell layer thickness induced by FasL in PACAP+/+ and PACAP-/- mice. Second, ex vivo and immunohistochemical studies revealed that interaction between FasL and PACAP occurs through the caspase 3 activity. Third, behavioural study showed a significant difference for the PACAP + FasL group in the righting reflex test at P8 which does not persist at P60. Finally, a time course study revealed that the pro-apoptotic effect of FasL characterized at P8 was followed by a progressive compensatory mechanism in caspase 3 activity and bromodeoxyuridine incorporation. These data suggest that PACAP and FasL interact during cerebellar development to control apoptosis of granule cells and may affect some motor cerebellar functions.

Published
2010
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18. Molecular and conformational determinants of pituitary adenylate cyclase-activating polypeptide (PACAP) for activation of the PAC1 receptor

Author

David Vaudry, Steve Bourgault, Marc Laburthe, Isabelle Ségalas-Milazzo, Alain Couvineau, Laure Guilhaudis, Alain Fournier, Hubert Vaudry, Neuroendocrinologie cellulaire et moléculaire, Université de Rouen Normandie (UNIROUEN), Normandie Université (NU)-Normandie Université (NU)-Institut National de la Santé et de la Recherche Médicale (INSERM), Institut Fédératif de Recherches Multidisciplinaires sur les Peptides (IFRMP 23), Institut National de la Santé et de la Recherche Médicale (INSERM)-Centre de Lutte Contre le Cancer Henri Becquerel Normandie Rouen (CLCC Henri Becquerel)-Université de Rouen Normandie (UNIROUEN), Normandie Université (NU)-Normandie Université (NU)-Université Le Havre Normandie (ULH), Normandie Université (NU)-CHU Rouen, Normandie Université (NU)-Centre National de la Recherche Scientifique (CNRS), Institut Armand Frappier (INRS-IAF), Institut National de la Recherche Scientifique [Québec] (INRS)-Réseau International des Instituts Pasteur (RIIP), International Associated laboratory Samuel de Champlain, Institut National de la Santé et de la Recherche Médicale (INSERM)-Institut National de la Recherche Scientifique [Québec] (INRS)-Université de Rouen Normandie (UNIROUEN), Normandie Université (NU)-Normandie Université (NU), Chimie Organique et Bioorganique : Réactivité et Analyse (COBRA), Institut Normand de Chimie Moléculaire Médicinale et Macromoléculaire (INC3M), Institut de Chimie du CNRS (INC)-École Nationale Supérieure d'Ingénieurs de Caen (ENSICAEN), Normandie Université (NU)-Normandie Université (NU)-Institut national des sciences appliquées Rouen Normandie (INSA Rouen Normandie), Institut National des Sciences Appliquées (INSA)-Normandie Université (NU)-Institut National des Sciences Appliquées (INSA)-Université Le Havre Normandie (ULH), Normandie Université (NU)-Université de Rouen Normandie (UNIROUEN), Normandie Université (NU)-Centre National de la Recherche Scientifique (CNRS)-Université de Caen Normandie (UNICAEN), Normandie Université (NU)-Institut de Chimie du CNRS (INC)-École Nationale Supérieure d'Ingénieurs de Caen (ENSICAEN), Normandie Université (NU)-Centre National de la Recherche Scientifique (CNRS)-Institut de Chimie Organique Fine (IRCOF), Institut National des Sciences Appliquées (INSA)-Normandie Université (NU)-Institut National des Sciences Appliquées (INSA)-Centre National de la Recherche Scientifique (CNRS)-Centre National de la Recherche Scientifique (CNRS), Equipe de Chimie Organique et Biologie Structurale (ECOBS), Institut national des sciences appliquées Rouen Normandie (INSA Rouen Normandie), Institut National des Sciences Appliquées (INSA)-Normandie Université (NU)-Institut National des Sciences Appliquées (INSA)-Normandie Université (NU), Centre de recherche biomédicale Bichat-Beaujon (CRB3), Université Paris Diderot - Paris 7 (UPD7)-Institut National de la Santé et de la Recherche Médicale (INSERM), and This work was supported by the Institut National de la Recherche Scientifique, the Institut National de la Santé et de la Recherche Médicale (INSERM U413), the Conseil Régional de Haute-Normandie, the IREB, the IRME, the ANR, and a scientific exchange program from INSERM and the Fonds de la Recherche en Sante' du Que'bec (FRSQ). Parts of these studies were performed with the support of the Platform for Cell Imaging of Haute-Normandie (PRIMACEN). NMR and molecular modeling software facilities were provided by the Centre de Ressources Informatiques de Haute-Normandie

Subjects
Agonist, Magnetic Resonance Spectroscopy, MESH: Amino Acids, Protein Conformation, medicine.drug_class, Adenylate kinase, MESH: Cricetinae, CHO Cells, Cyclase, Turn (biochemistry), Structure-Activity Relationship, 03 medical and health sciences, chemistry.chemical_compound, Cricetulus, 0302 clinical medicine, MESH: Structure-Activity Relationship, MESH: Protein Conformation, MESH: Cricetulus, MESH: CHO Cells, Cricetinae, Drug Discovery, Peptide synthesis, medicine, Animals, MESH: Protein Binding, MESH: Animals, Amino Acids, 030304 developmental biology, G protein-coupled receptor, 0303 health sciences, MESH: Magnetic Resonance Spectroscopy, MESH: Pituitary Adenylate Cyclase-Activating Polypeptide, Pituitary adenylate cyclase-activating peptide, chemistry, Biochemistry, [SDV.TOX]Life Sciences [q-bio]/Toxicology, MESH: Receptors, Pituitary Adenylate Cyclase-Activating Polypeptide, Type I, Pituitary Adenylate Cyclase-Activating Polypeptide, Molecular Medicine, Pharmacophore, 030217 neurology & neurosurgery, Protein Binding, Receptors, Pituitary Adenylate Cyclase-Activating Polypeptide, Type I
Abstract

International audience; PAC1 receptor is abundant in the CNS and plays an important role in neuronal survival. To identify the molecular determinants and the conformational components responsible for the activation of the PAC1 receptor, we performed a SAR study focusing on the N-terminal domain of its endogenous ligand, PACAP. This approach revealed that residues Asp(3) and Phe(6) are key elements of the pharmacophore of the PAC1 receptor. This study, supported by NMR structural analyses, suggests that the N-terminal tail of PACAP (residues 1 to 4) adopts a specific conformation similar to a turn when it activates the PAC1 receptor. Moreover, the integrity of the alpha-helix conformation observed at positions 5 to 7 appears crucial to allow the binding of PACAP. Characterization of analogues led to the identification of several superagonists, such as [Bip(6)]PACAP27, and of a new potent PAC1 receptor antagonist, [Sar(4)]PACAP38. The bioactive conformation inferred from this SAR study could constitute an appropriate molecular scaffold supporting the design of nonpeptidic PAC1 receptor agonists.

Published
2009
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19. Distribution and functional characterization of pituitary adenylate cyclase-activating polypeptide receptors in the brain of non-human primates

Author

Nicolas Aubert, Marc Massonneau, M. Basille, J. F. Le Bigot, Alain Fournier, S. de Jouffrey, David Vaudry, Hubert Vaudry, Pauline Noack, Valérie Jolivel, Philippe Ancian, Bruno J. Gonzalez, Neuroendocrinologie cellulaire et moléculaire, Université de Rouen Normandie (UNIROUEN), Normandie Université (NU)-Normandie Université (NU)-Institut National de la Santé et de la Recherche Médicale (INSERM), Quantitative Imaging in Drug Development (QUIDD), QUantitative Imaging in Drug Development, International Associated laboratory Samuel de Champlain, Institut National de la Santé et de la Recherche Médicale (INSERM)-Institut National de la Recherche Scientifique [Québec] (INRS)-Université de Rouen Normandie (UNIROUEN), Normandie Université (NU)-Normandie Université (NU), Centre International de Toxicologie (CIT), Centre International de Toxicologie, Institut Armand Frappier (INRS-IAF), Institut National de la Recherche Scientifique [Québec] (INRS)-Réseau International des Instituts Pasteur (RIIP), Ce travail a été financé par l'Institut National de la Santé et de la Recherche Médicale (INSERM U-413), the Institut pour la Recherche sur la Moelle épinière et l'Encéphale (IRME), the Institut de Recherches Scientifiques sur les Boissons (IREB, and 2007/29), the Institut National de la Santé et de la Recherche Médicale and Fondation de la Recherche en Santé du Québec (INSERM/FRSQ

Subjects
Male, Vasoactive intestinal peptide, MESH: Callithrix, Receptors, Pituitary Adenylate Cyclase-Activating Polypeptide, 0302 clinical medicine, Tissue Distribution, MESH: Animals, Receptor, MESH: Brain Mapping, Brain Mapping, 0303 health sciences, General Neuroscience, Brain, Marmoset, Callithrix, Cell biology, Pituitary adenylate cyclase-activating peptide, medicine.anatomical_structure, Cerebral cortex, Hypothalamus, [SDV.TOX]Life Sciences [q-bio]/Toxicology, MESH: Habenula, Pituitary Adenylate Cyclase-Activating Polypeptide, Female, medicine.medical_specialty, Adenylate kinase, In situ hybridization, Biology, 03 medical and health sciences, MESH: Brain, Species Specificity, Internal medicine, biology.animal, medicine, Animals, MESH: Species Specificity, RNA, Messenger, MESH: Tissue Distribution, 030304 developmental biology, MESH: RNA, Messenger, Habenula, MESH: Receptors, Pituitary Adenylate Cyclase-Activating Polypeptide, MESH: Septum of Brain, MESH: Pituitary Adenylate Cyclase-Activating Polypeptide, MESH: Hypothalamus, MESH: Male, Macaca fascicularis, Endocrinology, MESH: Macaca fascicularis, Septum of Brain, MESH: Female, 030217 neurology & neurosurgery
Abstract

International audience; The distribution and density of pituitary adenylate cyclase-activating polypeptide (PACAP) binding sites have been investigated in the brain of the primates Jacchus callithrix (marmoset) and Macaca fascicularis (macaque) using [(125)I]-PACAP27 as a radioligand. PACAP binding sites were widely expressed in the brain of these two species with particularly high densities in the septum, hypothalamus and habenula. A moderate density of recognition sites was seen in all subdivisions of the cerebral cortex with a heterogenous distribution, the highest concentrations occurring in layers I and VI while the underlying white matter was almost devoid of binding sites. Reverse transcriptase-polymerase chain reaction (RT-PCR) analysis revealed intense expression of the mRNAs encoding the short and hop-1 variants of pituitary adenylate cyclase-activating polypeptide-specific receptor (PAC1-R) in the cortex of both marmoset and macaque, whereas vasoactive intestinal polypeptide/pituitary adenylate cyclase-activating polypeptide mutual receptor, subtype 1 (VPAC1-R) and vasoactive intestinal polypeptide/pituitary adenylate cyclase-activating polypeptide mutual receptor, subtype 2 (VPAC2-R) mRNAs were expressed at a much lower level. In situ hybridization histochemistry showed intense expression of PAC1-R and weak expression of VPAC1-R mRNAs in layer IV of the cerebral cortex. Incubation of cortical tissue slices with PACAP induced a dose-dependent stimulation of cyclic AMP formation, indicating that PACAP binding sites correspond to functional receptors. Moreover, treatment of primate cortical slices with 100 nM PACAP significantly reduced the activity of caspase-3, a key enzyme of the apoptotic cascade. The present results indicate that PACAP should exert the same neuroprotective effect in the brain of primates as in rodents and suggest that PAC1-R agonists may have a therapeutic value to prevent neuronal cell death after stroke or in specific neurodegenerative diseases.

Published
2009
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20. Biological and structural analysis of truncated analogs of PACAP27

Author

Hubert Vaudry, Emilie Raoult, Steve Bourgault, Alain Couvineau, Laure Guilhaudis, Alain Fournier, Isabelle Ségalas-Milazzo, David Vaudry, Marc Laburthe, Institut Armand Frappier (INRS-IAF), Institut National de la Recherche Scientifique [Québec] (INRS)-Réseau International des Instituts Pasteur (RIIP), Neuroendocrinologie cellulaire et moléculaire, Université de Rouen Normandie (UNIROUEN), Normandie Université (NU)-Normandie Université (NU)-Institut National de la Santé et de la Recherche Médicale (INSERM), International Associated laboratory Samuel de Champlain, Institut National de la Santé et de la Recherche Médicale (INSERM)-Institut National de la Recherche Scientifique [Québec] (INRS)-Université de Rouen Normandie (UNIROUEN), Normandie Université (NU)-Normandie Université (NU), Chimie Organique et Bioorganique : Réactivité et Analyse (COBRA), Institut Normand de Chimie Moléculaire Médicinale et Macromoléculaire (INC3M), Institut de Chimie du CNRS (INC)-École Nationale Supérieure d'Ingénieurs de Caen (ENSICAEN), Normandie Université (NU)-Normandie Université (NU)-Institut national des sciences appliquées Rouen Normandie (INSA Rouen Normandie), Institut National des Sciences Appliquées (INSA)-Normandie Université (NU)-Institut National des Sciences Appliquées (INSA)-Université Le Havre Normandie (ULH), Normandie Université (NU)-Université de Rouen Normandie (UNIROUEN), Normandie Université (NU)-Centre National de la Recherche Scientifique (CNRS)-Université de Caen Normandie (UNICAEN), Normandie Université (NU)-Institut de Chimie du CNRS (INC)-École Nationale Supérieure d'Ingénieurs de Caen (ENSICAEN), Normandie Université (NU)-Centre National de la Recherche Scientifique (CNRS)-Institut de Chimie Organique Fine (IRCOF), Institut National des Sciences Appliquées (INSA)-Normandie Université (NU)-Institut National des Sciences Appliquées (INSA)-Centre National de la Recherche Scientifique (CNRS)-Centre National de la Recherche Scientifique (CNRS), Centre de recherche biomédicale Bichat-Beaujon (CRB3), Université Paris Diderot - Paris 7 (UPD7)-Institut National de la Santé et de la Recherche Médicale (INSERM), and This work was supported by the Agence Nationale de la Recherche (ANR), the Institut National de la Recherche Scientifique, the Institut National de la Santé et de la Recherche Médicale (INSERM U413), the Conseil Régional de Haute-Normandie, and a scientific exchange program from INSERM and the Fonds de la Recherche en Santé du Québec. Parts of these studies were performed with the support of the Platform for Cell Imaging of Haute-Normandie. H.V. is an Affiliated Professor at the Institut National de la Recherche Scientifique--Institut Armand-Frappier.

Subjects
Circular dichroism, MESH: Rats, Molecular Sequence Data, MESH: Cricetinae, MESH: Amino Acid Sequence, CHO Cells, PAC1 Receptor, PC12 Cells, MESH: Circular Dichroism, 03 medical and health sciences, Cellular and Molecular Neuroscience, 0302 clinical medicine, Cricetulus, MESH: Cricetulus, MESH: CHO Cells, MESH: Cell Proliferation, Cricetinae, MESH: PC12 Cells, Animals, MESH: Animals, Amino Acid Sequence, Receptor, MESH: Peptide Fragments, Peptide sequence, 030304 developmental biology, Cell Proliferation, 0303 health sciences, MESH: Molecular Sequence Data, biology, Chemistry, Cell growth, Chinese hamster ovary cell, Circular Dichroism, MESH: Pituitary Adenylate Cyclase-Activating Polypeptide, Biological activity, General Medicine, biology.organism_classification, Peptide Fragments, Rats, Biochemistry, [SDV.TOX]Life Sciences [q-bio]/Toxicology, MESH: Receptors, Pituitary Adenylate Cyclase-Activating Polypeptide, Type I, Biophysics, Pituitary Adenylate Cyclase-Activating Polypeptide, 030217 neurology & neurosurgery, Receptors, Pituitary Adenylate Cyclase-Activating Polypeptide, Type I
Abstract

International audience; The affinity toward the PAC1 receptor, the biological activity, and the alpha-helical content of several truncated PACAP27 analogs were measured. We first evaluated the pharmacological and structural parameters of C-terminal shortened PACAP fragments, from PACAP(1-23) to PACAP(1-19). All carboxy-truncated derivatives demonstrated circular dichroism spectra typical of a helical conformation. On the other hand, progressive shortening of the C-terminal domain gradually decreases the potency of PACAP to bind and to activate the PAC1 receptor. This decrease in biological activity was mainly attributed to the removal of residues that seem to interact directly with the receptor rather than to a destabilization of the C-terminal helical conformation. We also investigated the pharmacological and conformational characteristics of several hybrid PACAP27 derivatives containing an aliphatic molecular spacer connecting the N-terminal domain to the C-terminal region. However, this strategy revealed that none of these discontinuous analogs showed any significant affinity toward the PAC1 receptor, even if some of them exhibited circular dichroism spectra corresponding to an alpha-helical structure. This study suggests that several domains of PACAP27 are involved in the interaction with the PAC1 receptor and that the presence of the helical conformation is not a sufficient feature for receptor activation.

Published
2008
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21. Selenoprotein T is a PACAP-regulated gene involved in intracellular Ca2+ mobilization and neuroendocrine secretion

Author

Nathalie C. Guérineau, Alain Fournier, Didier Vieau, Hubert Vaudry, Youssef Anouar, Stéphane Gasman, Abdel G. Elkahloun, Luca Grumolato, Djida Ait-Ali, Jean Lesage, Ludovic Galas, Hafida Ghzili, Yannick Tanguy, Maité Montero-Hadjadje, Jérôme Leprince, Neuroendocrinologie cellulaire et moléculaire, Université de Rouen Normandie (UNIROUEN), Normandie Université (NU)-Normandie Université (NU)-Institut National de la Santé et de la Recherche Médicale (INSERM), Différenciation et communication neuronale et neuroendocrine (DC2N), Neurotransmission et sécrétion neuroendocrine (NSN), Centre National de la Recherche Scientifique (CNRS), Environnement périnatal et croissance - EA 4489 (EPS), Université de Lille-Centre Hospitalier Régional Universitaire [Lille] (CHRU Lille), Plate-Forme de Recherche en Imagerie Cellulaire de Haute-Normandie (PRIMACEN), Normandie Université (NU)-Normandie Université (NU)-Institute for Research and Innovation in Biomedicine (IRIB), Normandie Université (NU)-Normandie Université (NU)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Centre National de la Recherche Scientifique (CNRS)-Université de Rouen Normandie (UNIROUEN), Normandie Université (NU)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Centre National de la Recherche Scientifique (CNRS)-High-tech Research Infrastructures for Life Sciences (HeRacLeS), Normandie Université (NU)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Centre National de la Recherche Scientifique (CNRS)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Centre National de la Recherche Scientifique (CNRS)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Centre National de la Recherche Scientifique (CNRS), Institut de Génomique Fonctionnelle (IGF), Université Montpellier 1 (UM1)-Université Montpellier 2 - Sciences et Techniques (UM2)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Centre National de la Recherche Scientifique (CNRS), Cancer Genetics Branch, National Institute of Health (NIH)-National Human Genome Research Institute (NHGRI), Montreal Heart Institute (MONTREAL HEART INSTITUTE), Normandie Université (NU)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Centre National de la Recherche Scientifique (CNRS), Université de Montpellier (UM)-Université Montpellier 1 (UM1)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Université Montpellier 2 - Sciences et Techniques (UM2)-Centre National de la Recherche Scientifique (CNRS), Normandie Université (NU)-Normandie Université (NU)-Centre National de la Recherche Scientifique (CNRS)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Université de Rouen Normandie (UNIROUEN), and Normandie Université (NU)-Centre National de la Recherche Scientifique (CNRS)-Institut National de la Santé et de la Recherche Médicale (INSERM)

Subjects
[SDV.NEU.NB]Life Sciences [q-bio]/Neurons and Cognition [q-bio.NC]/Neurobiology, Cellular differentiation, [CHIM.THER]Chemical Sciences/Medicinal Chemistry, Endoplasmic Reticulum, Biochemistry, PC12 Cells, MESH: Selenoproteins, [SDV.BC.IC]Life Sciences [q-bio]/Cellular Biology/Cell Behavior [q-bio.CB], Gene expression, Cyclic AMP, MESH: Animals, Selenoproteins, MESH: Cyclic AMP, Regulation of gene expression, chemistry.chemical_classification, 0303 health sciences, Gene knockdown, Neurosecretion, 030302 biochemistry & molecular biology, Selenoprotein T, Cell Differentiation, MESH: Gene Expression Regulation, MESH: Calcium, Pituitary Adenylate Cyclase-Activating Polypeptide, Biotechnology, MESH: Cell Differentiation, MESH: Selenocysteine, MESH: Rats, MESH: Neurosecretion, Biology, MESH: Calcium Signaling, 03 medical and health sciences, MESH: Endoplasmic Reticulum, Genetics, MESH: PC12 Cells, Animals, Secretion, Calcium Signaling, Molecular Biology, 030304 developmental biology, Endoplasmic reticulum, MESH: Pituitary Adenylate Cyclase-Activating Polypeptide, Molecular biology, Rats, Selenocysteine, chemistry, Gene Expression Regulation, [SDV.SP.PHARMA]Life Sciences [q-bio]/Pharmaceutical sciences/Pharmacology, Calcium, Selenoprotein
Abstract

International audience; Selenoproteins contain the essential trace element selenium, the deficiency of which is associated with cancer or accelerated aging. Although selenoproteins are thought to be instrumental for the effects of selenium, the biological function of many of these proteins remains unknown. Here, we studied the role of selenoprotein T (SelT), a selenocysteine (Sec) -containing protein with no known function, which we have identified as a novel target gene of the neuropeptide pituitary adenylate cyclase-activating polypeptide (PACAP) during PC12 cell differentiation. SelT was found to be ubiquitously expressed throughout embryonic development and in adulthood in rat. Immunocytochemical analysis revealed that SelT is mainly localized to the endoplasmic reticulum through a hydrophobic domain. PACAP and cAMP induced a rapid and long-lasting increase in SelT gene expression in PC12 cells, in a Ca(2+)-dependent manner. These results suggested a possible role of SelT in PACAP signaling during PC12 cell differentiation. Indeed, overexpression of SelT in PC12 cells provoked an increase in the concentration of intracellular Ca(2+) ([Ca(2+)](i)) that was dependent on the Sec residue. Conversely, SelT gene knockdown inhibited the PACAP-induced increase in [Ca(2+)](i) and reduced hormone secretion. These findings demonstrate the implication of a selenoprotein in the regulation of Ca(2+) homeostasis and neuroendocrine secretion in response to a cAMP-stimulating trophic factor.

Published
2008
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22. Role of PACAP and VIP in astroglial functions

Author

Jérôme Leprince, Alain Fournier, Pierrick Gandolfo, Hélène Castel, Marie-Christine Tonon, Agnieszka Dejda, Olfa Masmoudi-Kouki, Hubert Vaudry, Neuroendocrinologie cellulaire et moléculaire, Université de Rouen Normandie (UNIROUEN), Normandie Université (NU)-Normandie Université (NU)-Institut National de la Santé et de la Recherche Médicale (INSERM), Institut Fédératif de Recherches Multidisciplinaires sur les Peptides (IFRMP 23), Institut National de la Santé et de la Recherche Médicale (INSERM)-Centre de Lutte Contre le Cancer Henri Becquerel Normandie Rouen (CLCC Henri Becquerel)-Université de Rouen Normandie (UNIROUEN), Normandie Université (NU)-Normandie Université (NU)-Université Le Havre Normandie (ULH), Normandie Université (NU)-CHU Rouen, Normandie Université (NU)-Centre National de la Recherche Scientifique (CNRS), Faculté des Sciences Mathématiques, Physiques et Naturelles de Tunis (FST), Université de Tunis El Manar (UTM), International Associated laboratory Samuel de Champlain, Institut National de la Recherche Scientifique [Québec] (INRS)-Université de Rouen Normandie (UNIROUEN), Institut Armand Frappier (INRS-IAF), Institut National de la Recherche Scientifique [Québec] (INRS)-Réseau International des Instituts Pasteur (RIIP), and Polish Academy of Sciences (PAN)

Subjects
Physiology, MESH: Receptors, Vasoactive Intestinal Peptide, [SDV.NEU.NB]Life Sciences [q-bio]/Neurons and Cognition [q-bio.NC]/Neurobiology, Vasoactive intestinal peptide, MESH: Glycogen, Receptors, Pituitary Adenylate Cyclase-Activating Polypeptide, Neuropeptide, [CHIM.THER]Chemical Sciences/Medicinal Chemistry, Biology, Biochemistry, Neuroprotection, Models, Biological, 03 medical and health sciences, Cellular and Molecular Neuroscience, 0302 clinical medicine, Endocrinology, Neurotrophic factors, MESH: Cell Proliferation, [SDV.BC.IC]Life Sciences [q-bio]/Cellular Biology/Cell Behavior [q-bio.CB], medicine, Animals, Humans, MESH: Animals, 030304 developmental biology, Cell Proliferation, 0303 health sciences, MESH: Receptors, Pituitary Adenylate Cyclase-Activating Polypeptide, MESH: Humans, MESH: Pituitary Adenylate Cyclase-Activating Polypeptide, MESH: Vasoactive Intestinal Peptide, MESH: Models, Biological, MESH: Astrocytes, Pituitary adenylate cyclase-activating peptide, medicine.anatomical_structure, Astrocytes, Synaptic plasticity, [SDV.SP.PHARMA]Life Sciences [q-bio]/Pharmaceutical sciences/Pharmacology, Neuroglia, Pituitary Adenylate Cyclase-Activating Polypeptide, Receptors, Vasoactive Intestinal Peptide, Neuroscience, 030217 neurology & neurosurgery, Glycogen, hormones, hormone substitutes, and hormone antagonists, Astrocyte, Vasoactive Intestinal Peptide
Abstract

International audience; Astrocytes represent at least 50% of the volume of the human brain. Besides their roles in various supportive functions, astrocytes are involved in the regulation of stem cell proliferation, synaptic plasticity and neuroprotection. Astrocytes also influence neuronal physiology by responding to neurotransmitters and neuropeptides and by releasing regulatory factors termed gliotransmitters. In particular, astrocytes express the PACAP-specific receptor PAC1-R and the PACAP/VIP mutual receptors VPAC1-R and VPAC2-R during development and/or in the adult. There is now clear evidence that PACAP and VIP modulate a number of astrocyte activities such as proliferation, plasticity, glycogen production, and biosynthesis of neurotrophic factors and gliotransmitters.

Published
2007
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23. VIP and PACAP stimulate TSH release from the bullfrog pituitary

Author

Alain Fournier, Kazutoshi Yamamoto, Sakae Kikuyama, Reiko Okada, Marie Christine Tonon, Jérôme Leprince, Hubert Vaudry, Hiroshi Mochida, Yoichi Ito, Waseda University, Saitama University, University of Shizuoka, Neuroendocrinologie cellulaire et moléculaire, Université de Rouen Normandie (UNIROUEN), Normandie Université (NU)-Normandie Université (NU)-Institut National de la Santé et de la Recherche Médicale (INSERM), International Associated laboratory Samuel de Champlain, Institut National de la Santé et de la Recherche Médicale (INSERM)-Institut National de la Recherche Scientifique [Québec] (INRS)-Université de Rouen Normandie (UNIROUEN), Normandie Université (NU)-Normandie Université (NU), Institut Armand Frappier (INRS-IAF), and Institut National de la Recherche Scientifique [Québec] (INRS)-Réseau International des Instituts Pasteur (RIIP)

Subjects
Pituitary gland, Physiology, [SDV.NEU.NB]Life Sciences [q-bio]/Neurons and Cognition [q-bio.NC]/Neurobiology, Vasoactive intestinal peptide, Receptors, Pituitary Adenylate Cyclase-Activating Polypeptide, Thyrotropin, [CHIM.THER]Chemical Sciences/Medicinal Chemistry, PACAP, Biochemistry, MESH: Dose-Response Relationship, Drug, 0302 clinical medicine, Endocrinology, Thyrotropic cell, Bullfrog, MESH: Pituitary Gland, [SDV.BC.IC]Life Sciences [q-bio]/Cellular Biology/Cell Behavior [q-bio.CB], MESH: Animals, Cells, Cultured, 0303 health sciences, Rana catesbeiana, Chemistry, MESH: Vasoactive Intestinal Peptide, PAC1-receptor, Pituitary adenylate cyclase-activating peptide, medicine.anatomical_structure, Pituitary Gland, MESH: Thyrotropin, Pituitary Adenylate Cyclase-Activating Polypeptide, Frog TSH, hormones, hormone substitutes, and hormone antagonists, Endocrine gland, Vasoactive Intestinal Peptide, MESH: Cells, Cultured, medicine.medical_specialty, endocrine system, 03 medical and health sciences, Cellular and Molecular Neuroscience, Thyroid-stimulating hormone, Internal medicine, medicine, Animals, 030304 developmental biology, MESH: Receptors, Pituitary Adenylate Cyclase-Activating Polypeptide, Dose-Response Relationship, Drug, MESH: Pituitary Adenylate Cyclase-Activating Polypeptide, Antagonist, MESH: Rana catesbeiana, VIP, VPAC2-receptor, [SDV.SP.PHARMA]Life Sciences [q-bio]/Pharmaceutical sciences/Pharmacology, 030217 neurology & neurosurgery
Abstract

International audience; We have recently shown that corticotropin-releasing hormone (CRH) is a major thyrotropin (TSH)-releasing factor in amphibians, but we have also found that, besides CRH, other hypothalamic substances stimulate TSH secretion in frog. In order to characterize novel TSH secretagogues, we have investigated the effect of frog (Rana ridibunda) vasoactive intestinal polypeptide (VIP) (fVIP) and pituitary adenylate cyclase-activating polypeptide (PACAP) (fPACAP38 and PACAP27) on TSH release from bullfrog (Rana catesbeiana) pituitary cells in primary culture. Incubation of pituitary cells for 24h with graded concentrations of fVIP, fPACAP38 and PACAP27 (10(-9) to 10(-6)M) induced a dose-dependent stimulation of TSH release with minimum effective doses of 10(-9)M for fVIP and 10(-8)M for fPACAP38 and PACAP27. The PAC1-R/VPAC2-R antagonist PACAP(6-38) (10(-7) and 10(-6)M) dose-dependently suppressed the stimulatory effects of fVIP and fPACAP38 (10(-7)M each). Likewise, this antagonist (10(-6) and 10(-5)M) dose-dependently attenuated the stimulatory effect of PACAP27 (10(-7)M). On the other hand, the VPAC1-R/VPAC2-R antagonist [d-pCl-Phe(6), Leu(17)]VIP (10(-6) and 10(-5)M) dose-dependently inhibited the stimulatory effect of fVIP (10(-9)M) and PACAP27 (10(-8)M), but did not affect the response to fPACAP38 (10(-8)M). These data indicate that, in amphibians, the activity of thyrotrophs can be regulated by VIP and PACAP acting likely through VPAC2-R and PAC1-R.

Published
2007
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24. Actions of PACAP and VIP on melanotrope cells of Xenopus laevis

Author

Kidane, Adhanet, Cruijsen, Peter, Ortiz-Bazan, Maria, Vaudry, Hubert, Leprince, Jérôme, Kuijpers-Kwant, Frouwke, Roubos, Eric, Jenks, Bruce, Cruijsen, Peter M.J.M., Neuroendocrinologie cellulaire et moléculaire, Université de Rouen Normandie (UNIROUEN), Normandie Université (NU)-Normandie Université (NU)-Institut National de la Santé et de la Recherche Médicale (INSERM), and Différenciation et communication neuronale et neuroendocrine (DC2N)

Subjects
Pro-Opiomelanocortin, Physiology, [SDV.NEU.NB]Life Sciences [q-bio]/Neurons and Cognition [q-bio.NC]/Neurobiology, Vasoactive intestinal peptide, Xenopus, Gene Expression, Enteroendocrine cell, [CHIM.THER]Chemical Sciences/Medicinal Chemistry, Biochemistry, MESH: Dose-Response Relationship, Drug, Xenopus laevis, 0302 clinical medicine, Endocrinology, MESH: Pituitary Gland, MESH: Reverse Transcriptase Polymerase Chain Reaction, [SDV.BC.IC]Life Sciences [q-bio]/Cellular Biology/Cell Behavior [q-bio.CB], MESH: Pro-Opiomelanocortin, MESH: Animals, Receptor, Regulation of gene expression, 0303 health sciences, Reverse Transcriptase Polymerase Chain Reaction, MESH: Vasoactive Intestinal Peptide, Pars intermedia, Immunohistochemistry, 3. Good health, Pituitary adenylate cyclase-activating peptide, Pituitary Gland, Pituitary Adenylate Cyclase-Activating Polypeptide, hormones, hormone substitutes, and hormone antagonists, Vasoactive Intestinal Peptide, medicine.medical_specialty, endocrine system, MESH: Gene Expression, Melanotrophs, Neurophysiology, Neuropeptide, Biology, 03 medical and health sciences, Cellular and Molecular Neuroscience, MESH: Xenopus laevis, Internal medicine, medicine, Animals, 030304 developmental biology, Dose-Response Relationship, Drug, MESH: Pituitary Adenylate Cyclase-Activating Polypeptide, MESH: Immunohistochemistry, biology.organism_classification, MESH: Melanotrophs, [SDV.SP.PHARMA]Life Sciences [q-bio]/Pharmaceutical sciences/Pharmacology, 030217 neurology & neurosurgery
Abstract

Item does not contain fulltext The neuropeptides, pituitary adenylate cyclase-activating polypeptide (PACAP) and vasoactive intestinal polypeptide (VIP) are implicated in the regulation of gene expression and hormone secretion in mammalian melanotrope cells and a mammalian pro-opiomelanocortin (POMC)-producing tumor cell line, but the physiological relevance of this regulation is elusive. The purpose of the present study was to establish if these peptides affect biosynthetic and secretory processes in a well-established physiological model for endocrine cell functioning, the pituitary melanotrope cells of the amphibian Xenopus laevis, which hormonally control the process of skin color adaptation to background illumination. We show that both PACAP and VIP are capable of stimulating the secretory process of the Xenopus melanotrope cell. As the peptides are equipotent, they may exert their actions via a VPAC receptor. Moreover, PACAP stimulated POMC biosynthesis and POMC gene expression. Strong anti-PACAP immunoreactivity was found in the pituitary pars nervosa (PN), suggesting that this neurohemal organ is a source of neurohormonal PACAP action on the melanotropes in the intermediate pituitary. We propose that the PACAP/VIP family of peptides has a physiological function in regulating Xenopus melanotrope cell activity during the process of skin color adaptation.

Published
2007
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25. PACAP and C2-ceramide generate different AP-1 complexes through a MAP-kinase-dependent pathway: involvement of c-Fos in PACAP-induced Bcl-2 expression

Author

Cecile Fisch, Xavier Xifró, Jean‐François Lebigot, Nicolas Aubert, Bruno J. Gonzalez, Alain Fournier, Hubert Vaudry, Stephane De Jouffrey, José Rodríguez-Alvarez, David Vaudry, Anthony Falluel-Morel, Neuroendocrinologie cellulaire et moléculaire, Université de Rouen Normandie (UNIROUEN), Normandie Université (NU)-Normandie Université (NU)-Institut National de la Santé et de la Recherche Médicale (INSERM), CIT (CIT), CIT, Laboratory of Biochemistry and Molecular Biology, Neuroscience Institute-Autonomous University of Barcelona, Institut Armand Frappier (INRS-IAF), Institut National de la Recherche Scientifique [Québec] (INRS)-Réseau International des Instituts Pasteur (RIIP), and This work was supported by INSERM (U413), the CIT, a France-Quebec exchange program (INSERM-FRSQ), the Regional Platform for Cell Imaging, and the Conseil Régional de Haute-Normandie

Subjects
Proto-Oncogene Proteins c-jun, MESH: Neurons, Apoptosis, Biochemistry, c-Fos, MESH: Animals, Newborn, MESH: Down-Regulation, MESH: Dose-Response Relationship, Drug, 0302 clinical medicine, Sphingosine, Gene expression, Phosphoprotein Phosphatases, MESH: Animals, Protein Phosphatase 2, Enzyme Inhibitors, Phosphorylation, Cells, Cultured, Neurons, 0303 health sciences, Kinase, MESH: Proto-Oncogene Proteins c-fos, MESH: Protein Phosphatase 2, MESH: Gene Expression Regulation, MESH: Transcription Factor AP-1, 3. Good health, DNA-Binding Proteins, Proto-Oncogene Proteins c-bcl-2, MESH: Cell Survival, MESH: Enzyme Inhibitors, Mitogen-activated protein kinase, [SDV.TOX]Life Sciences [q-bio]/Toxicology, Pituitary Adenylate Cyclase-Activating Polypeptide, MESH: Sphingosine, Proto-Oncogene Proteins c-fos, hormones, hormone substitutes, and hormone antagonists, MESH: Cells, Cultured, Transcriptional Activation, endocrine system, MESH: Rats, Cell Survival, MAP Kinase Signaling System, Down-Regulation, Adenylate kinase, Biology, Cerebellar Cortex, 03 medical and health sciences, Cellular and Molecular Neuroscience, MESH: Phosphoprotein Phosphatases, Animals, Rats, Wistar, Protein kinase A, 030304 developmental biology, MESH: Cerebellar Cortex, Dose-Response Relationship, Drug, MESH: Phosphorylation, MESH: Proto-Oncogene Proteins c-jun, MESH: MAP Kinase Signaling System, MESH: Apoptosis, MESH: Pituitary Adenylate Cyclase-Activating Polypeptide, Protein phosphatase 2, MESH: Rats, Wistar, Molecular biology, Rats, Transcription Factor AP-1, Animals, Newborn, Gene Expression Regulation, MESH: Proto-Oncogene Proteins c-bcl-2, biology.protein, MESH: Transcriptional Activation, 030217 neurology & neurosurgery, MESH: DNA-Binding Proteins
Abstract

International audience; The neuropeptide pituitary adenylate cyclase-activating polypeptide (PACAP) inhibits C2-ceramide-induced cell death through blockade of the mitochondrial apoptotic pathway in rat cerebellar granule neurones. However, the gene induction processes and transcription factors involved in the anti-apoptotic effect of PACAP remain unknown. Here, we show that PACAP and C2-ceramide activate activator protein-1 (AP-1) DNA binding in a dose- and time-dependent manner, but generate different AP-1 dimers. Thus, PACAP increased the proportion of c-Fos and Jun D while C2-ceramide increased c-Jun and reduced c-Fos in AP-1 complexes. In addition, PACAP strongly activated c-Fos gene expression while C2-ceramide markedly increased c-Jun phosphorylation. The effect of PACAP on c-Fos expression was blocked by the mitogen-activated protein kinase/extracellular signal-regulated kinase (MEK) inhibitor, U0126, while phosphorylation of c-Jun induced by C2-ceramide was abrogated by the protein phosphatase 2A (PP2A) inhibitor, okadaic acid. Transfection of immature granule cells with c-Fos siRNA, which strongly reduced basal and PACAP-stimulated levels of the protein, totally prevented the stimulatory effect of PACAP on Bcl-2 expression. The present study demonstrates that AP-1 complexes containing c-Fos mediate the effect of PACAP on Bcl-2 gene expression in cerebellar granule neurones. Our data also indicate that different AP-1 dimers are associated with the pro-apoptotic effect of C2-ceramide and the anti-apoptotic effect of PACAP.

Published
2006
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26. PACAP stimulates biosynthesis and release of endozepines from rat astrocytes

Author

Marie-Christine Tonon, Georges Pelletier, David Vaudry, Pierrick Gandolfo, Olfa Masmoudi-Kouki, Alain Fournier, Hubert Vaudry, Jérôme Leprince, Faculty of Sciences of Tunis (University of Tunis), Université de Tunis El Manar (UTM), Neuroendocrinologie cellulaire et moléculaire, Université de Rouen Normandie (UNIROUEN), Normandie Université (NU)-Normandie Université (NU)-Institut National de la Santé et de la Recherche Médicale (INSERM), Institut Fédératif de Recherches Multidisciplinaires sur les Peptides (IFRMP 23), Institut National de la Santé et de la Recherche Médicale (INSERM)-Centre de Lutte Contre le Cancer Henri Becquerel Normandie Rouen (CLCC Henri Becquerel)-Université de Rouen Normandie (UNIROUEN), Normandie Université (NU)-Normandie Université (NU)-Université Le Havre Normandie (ULH), Normandie Université (NU)-CHU Rouen, Normandie Université (NU)-Centre National de la Recherche Scientifique (CNRS), Laval University Medical center, Université Laval [Québec] (ULaval), Institut Armand Frappier (INRS-IAF), and Institut National de la Recherche Scientifique [Québec] (INRS)-Réseau International des Instituts Pasteur (RIIP)

Subjects
MESH: Signal Transduction, medicine.medical_specialty, endocrine system, Endozepine, MESH: Rats, [SDV.NEU.NB]Life Sciences [q-bio]/Neurons and Cognition [q-bio.NC]/Neurobiology, Vasoactive intestinal peptide, Neuropeptide, MESH: Adenylyl Cyclases, MESH: Cyclic AMP-Dependent Protein Kinases, [CHIM.THER]Chemical Sciences/Medicinal Chemistry, General Biochemistry, Genetics and Molecular Biology, Adenylyl cyclase, 03 medical and health sciences, chemistry.chemical_compound, 0302 clinical medicine, History and Philosophy of Science, Downregulation and upregulation, Internal medicine, [SDV.BC.IC]Life Sciences [q-bio]/Cellular Biology/Cell Behavior [q-bio.CB], medicine, Animals, MESH: Animals, Receptor, Protein kinase C, Cells, Cultured, 030304 developmental biology, Diazepam Binding Inhibitor, 0303 health sciences, Chemistry, General Neuroscience, MESH: Pituitary Adenylate Cyclase-Activating Polypeptide, Cyclic AMP-Dependent Protein Kinases, 3. Good health, Rats, MESH: Astrocytes, medicine.anatomical_structure, Endocrinology, Astrocytes, [SDV.SP.PHARMA]Life Sciences [q-bio]/Pharmaceutical sciences/Pharmacology, MESH: Diazepam Binding Inhibitor, Pituitary Adenylate Cyclase-Activating Polypeptide, 030217 neurology & neurosurgery, hormones, hormone substitutes, and hormone antagonists, Astrocyte, MESH: Cells, Cultured, Adenylyl Cyclases, Signal Transduction
Abstract

International audience; Astrocytes synthesize and release endozepines, a family of neuropeptides related to diazepam-binding inhibitor (DBI). Astroglial cells also express the receptors of pituitary adenylate cyclase-activating polypeptide (PACAP) and vasoactive intestinal polypeptide (VIP). In the present article, we show that PACAP dose dependently increases DBI gene expression and stimulates endozepine release through activation of PAC1-R. PACAP increases cAMP formation, enhances polyphosphoinositide turnover, and evokes calcium mobilization from intracellular Ca2+ pools. The effect of PACAP on endozepine release is mediated through the adenylyl cyclase/PKA pathway while the downregulation of astrocyte response to PACAP can be ascribed to activation of the PLC/PKC pathway.

Published
2006
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27. Identification of Proteins Regulated by PACAP in PC12 Cells by 2D Gel Electrophoresis Coupled to Mass Spectrometry

Author

Jérôme Leprince, Alexis Lebon, Laurent Coquet, Thierry Jouenne, Alain Fournier, Damien Seyer, Bruno J. Gonzalez, Pascal Cosette, David Vaudry, Hubert Vaudry, Philippe Chan, Plate-Forme de Recherche en Imagerie Cellulaire de Haute-Normandie (PRIMACEN), Université de Rouen Normandie (UNIROUEN), Normandie Université (NU)-Normandie Université (NU)-Institute for Research and Innovation in Biomedicine (IRIB), Normandie Université (NU)-Normandie Université (NU)-Centre National de la Recherche Scientifique (CNRS)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Université de Rouen Normandie (UNIROUEN), Normandie Université (NU)-Centre National de la Recherche Scientifique (CNRS)-Institut National de la Santé et de la Recherche Médicale (INSERM), Equipe de recherche sur les relations matrice extracellulaire-cellules (ERRMECe), Fédération INSTITUT DES MATÉRIAUX DE CERGY-PONTOISE (I-MAT), Université de Cergy Pontoise (UCP), Université Paris-Seine-Université Paris-Seine-Université de Cergy Pontoise (UCP), Université Paris-Seine-Université Paris-Seine, Polymères Biopolymères Surfaces (PBS), Institut national des sciences appliquées Rouen Normandie (INSA Rouen Normandie), Institut National des Sciences Appliquées (INSA)-Normandie Université (NU)-Institut National des Sciences Appliquées (INSA)-Normandie Université (NU)-Institut de Chimie du CNRS (INC)-Institut Normand de Chimie Moléculaire Médicinale et Macromoléculaire (INC3M), Institut de Chimie du CNRS (INC)-École Nationale Supérieure d'Ingénieurs de Caen (ENSICAEN), Normandie Université (NU)-Normandie Université (NU)-Institut national des sciences appliquées Rouen Normandie (INSA Rouen Normandie), Institut National des Sciences Appliquées (INSA)-Normandie Université (NU)-Institut National des Sciences Appliquées (INSA)-Université Le Havre Normandie (ULH), Normandie Université (NU)-Université de Rouen Normandie (UNIROUEN), Normandie Université (NU)-Centre National de la Recherche Scientifique (CNRS)-Université de Caen Normandie (UNICAEN), Normandie Université (NU)-École Nationale Supérieure d'Ingénieurs de Caen (ENSICAEN), Normandie Université (NU)-Université Le Havre Normandie (ULH), Normandie Université (NU)-Centre National de la Recherche Scientifique (CNRS), Polymères, biopolymères, membranes (PBM), Centre National de la Recherche Scientifique (CNRS)-Institut national des sciences appliquées Rouen Normandie (INSA Rouen Normandie), Institut National des Sciences Appliquées (INSA)-Normandie Université (NU)-Institut National des Sciences Appliquées (INSA)-Normandie Université (NU)-Université de Rouen Normandie (UNIROUEN), Normandie Université (NU), Neuroendocrinologie cellulaire et moléculaire, Normandie Université (NU)-Normandie Université (NU)-Institut National de la Santé et de la Recherche Médicale (INSERM), Différenciation et communication neuronale et neuroendocrine (DC2N), Montreal Heart Institute (MONTREAL HEART INSTITUTE), Endothélium microcirculatoire cérébral et lésions du système nerveux central au cours du développement (Néovasc), and Normandie Université (NU)-Centre National de la Recherche Scientifique (CNRS)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Institut National de la Santé et de la Recherche Médicale (INSERM)

Subjects
Neurite, MESH: Rats, Protein subunit, [SDV.NEU.NB]Life Sciences [q-bio]/Neurons and Cognition [q-bio.NC]/Neurobiology, Protein Array Analysis, PC12 cell line, [CHIM.THER]Chemical Sciences/Medicinal Chemistry, Proteomics, PC12 Cells, General Biochemistry, Genetics and Molecular Biology, 03 medical and health sciences, chemistry.chemical_compound, 0302 clinical medicine, History and Philosophy of Science, Hsp27, [SDV.BC.IC]Life Sciences [q-bio]/Cellular Biology/Cell Behavior [q-bio.CB], Animals, MESH: PC12 Cells, Electrophoresis, Gel, Two-Dimensional, MESH: Animals, MESH: Proteins, 030304 developmental biology, Gel electrophoresis, 0303 health sciences, Two-dimensional gel electrophoresis, biology, MESH: Protein Array Analysis, General Neuroscience, MESH: Pituitary Adenylate Cyclase-Activating Polypeptide, Proteins, MESH: Electrophoresis, Gel, Two-Dimensional, Molecular biology, Rats, Cell biology, Tubulin, chemistry, MESH: Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization, Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization, biology.protein, [SDV.SP.PHARMA]Life Sciences [q-bio]/Pharmaceutical sciences/Pharmacology, Pituitary Adenylate Cyclase-Activating Polypeptide, 030217 neurology & neurosurgery
Abstract

International audience; The rat pheochromocytoma PC12 cell line has been widely used as a model to study neuronal differentiation. In particular, after serum depletion, PC12 cells stop to proliferate and undergo apoptosis. Under such conditions, treatment with pituitary adenylate cyclase-activating polypeptide (PACAP) promotes cell survival and induces neurite outgrowth. The identification of the proteins regulated by PACAP in PC12 cells under apoptotic conditions should provide valuable information concerning the mechanisms controlling neuronal cell survival and differentiation. To this aim, PC12 cells cultured in serum-free medium were treated with PACAP (10(-7) M), proteins were extracted, separated by two-dimensional gel electrophoresis (2-DE), and identified by MALDI-ToF mass spectrometry. The comparison between 16 2-DE maps led to the characterization of 110 proteins regulated by PACAP among which 22 have been identified by automatic query of the Mascot, Aldente, and Profound servers with the ProGeR-CDD database. Seventy-six percent of these proteins, including the p17 subunit of caspase-3, the heat shock protein hsp60, and the GTPase ran were found to be repressed whereas the others notably hsp27, tubulin beta-5, and calmodulin were overexpressed. Investigation of the putative functions indicated that some of the proteins regulated by PACAP and identified in the present article could control cell survival or differentiation.

Published
2006
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28. Effects of pituitary adenylate cyclase-activating polypeptide, vasoactive intestinal polypeptide, and somatostatin on the release of thyrotropin from the bullfrog pituitary

Author

Sakae Kikuyama, Jérôme Leprince, Hubert Vaudry, Kazutoshi Yamamoto, Reiko Okada, Yoichi Ito, Alain Fournier, Nicolas Chartrel, Department of Biology, School of Education, Waseda University, Institut Fédératif de Recherches Multidisciplinaires sur les Peptides (IFRMP 23), Institut National de la Santé et de la Recherche Médicale (INSERM)-Centre de Lutte Contre le Cancer Henri Becquerel Normandie Rouen (CLCC Henri Becquerel)-Université de Rouen Normandie (UNIROUEN), Normandie Université (NU)-Normandie Université (NU)-Université Le Havre Normandie (ULH), Normandie Université (NU)-CHU Rouen, Normandie Université (NU)-Centre National de la Recherche Scientifique (CNRS), Institut Armand Frappier (INRS-IAF), Institut National de la Recherche Scientifique [Québec] (INRS)-Réseau International des Instituts Pasteur (RIIP), and This work was supported by Inamori Foundation (H17) JSPS (16370031, 15207007), Waseda University (2004B-839, 2003C-009), INSERM (U413), the European Institute for Peptide Research (IFRMP 23), and the Conseil Régional de Haute-Normandie.

Subjects
medicine.medical_specialty, endocrine system, endocrine system diseases, MESH: Somatostatin, Vasoactive intestinal peptide, Thyrotropin, Adenylate kinase, Neuropeptide, 030209 endocrinology & metabolism, Stimulation, somatostatin, PACAP, amphibia, General Biochemistry, Genetics and Molecular Biology, Cortistatin (neuropeptide), 03 medical and health sciences, 0302 clinical medicine, History and Philosophy of Science, Bullfrog, MESH: Pituitary Gland, Internal medicine, medicine, Animals, MESH: Animals, Cells, Cultured, 030304 developmental biology, 0303 health sciences, Rana catesbeiana, Chemistry, TSH, General Neuroscience, MESH: Pituitary Adenylate Cyclase-Activating Polypeptide, MESH: Vasoactive Intestinal Peptide, 3. Good health, MESH: Rana catesbeiana, VIP, Endocrinology, Somatostatin, Pituitary Gland, MESH: Thyrotropin, [SDV.TOX]Life Sciences [q-bio]/Toxicology, Pituitary Adenylate Cyclase-Activating Polypeptide, hormones, hormone substitutes, and hormone antagonists, Vasoactive Intestinal Peptide, Hormone, MESH: Cells, Cultured
Abstract

The recent development of a specific radioimmunoassay for amphibian (bullfrog, Rana catesbeiana) thyrotropin (TSH) has made it possible to study the effects of various neuropeptides on the release of TSH from the pituitary in vitro. Up to now, corticotropin-releasing factor of bullfrog origin has been shown to have a potent TSH-releasing activity, whereas gonadotropin-releasing hormone and TSH-releasing hormone exhibit a moderate TSH-releasing effect on the adult, but not larval, pituitary. In the present study, the effects of pituitary adenylate cyclase-activating polypeptide (PACAP), vasoactive intestinal polypeptide (VIP), and somatostatin (SS) on the in vitro release of TSH from the bullfrog pituitary were investigated. Both frog (R. ridibunda) PACAP-38 and PACAP-27 caused a concentration-dependent stimulation of the release of TSH from dispersed pituitary cells during a 24-h culture. The PACAP-38- and PACAP-27-induced TSH release was suppressed by a simultaneous application of PACAP6-38. Application of high concentrations of PACAP6-38 alone caused a slight but significant stimulatory effect on the release of TSH. Frog VIP also stimulated TSH release from pituitary cells concentration-dependently. Frog SS1 (homologous to mammalian somatostatin-14) and SS2 (homologous to mammalian cortistatin) did not affect the basal release of TSH but caused a concentration-dependent suppression of the PACAP-38-induced release of TSH. These results suggest the involvement of multiple neuropeptides in the regulation of the release of TSH from the amphibian pituitary.

Published
2006
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29. Opposite regulation of the mitochondrial apoptotic pathway by C2-ceramide and PACAP through a MAP-kinase-dependent mechanism in cerebellar granule cells

Author

Falluel-Morel, Anthony, Aubert, Nicolas, Vaudry, David, Basille, Magali, Fontaine, Marc, Fournier, Alain, Vaudry, Hubert, Gonzalez, Bruno J, Neuroendocrinologie cellulaire et moléculaire, Université de Rouen Normandie (UNIROUEN), Normandie Université (NU)-Normandie Université (NU)-Institut National de la Santé et de la Recherche Médicale (INSERM), Laboratory of Defense Proteins in Immune and Inflammatory Responses, Institut National de la Santé et de la Recherche Médicale (INSERM)-Faculty of Medicine and Pharmacy-European Institute for Peptide Research-Institut Fédératif de Recherches Multidisciplinaires sur les Peptides (IFRMP 23), Institut National de la Santé et de la Recherche Médicale (INSERM)-Centre de Lutte Contre le Cancer Henri Becquerel Normandie Rouen (CLCC Henri Becquerel)-Université de Rouen Normandie (UNIROUEN), Normandie Université (NU)-Normandie Université (NU)-Université Le Havre Normandie (ULH), Normandie Université (NU)-CHU Rouen, Normandie Université (NU)-Centre National de la Recherche Scientifique (CNRS)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Centre de Lutte Contre le Cancer Henri Becquerel Normandie Rouen (CLCC Henri Becquerel)-Université de Rouen Normandie (UNIROUEN), Normandie Université (NU)-Centre National de la Recherche Scientifique (CNRS), Institut Armand Frappier (INRS-IAF), Institut National de la Recherche Scientifique [Québec] (INRS)-Réseau International des Instituts Pasteur (RIIP), and This research was supported by an INSERM Grant (U413) and an IREB Grant (2001/22). AFM is recipient of a fellowship from the Conseil Re ́gional de Haute-Normandie (LARC-Neuroscience net- work). NA is a recipient of a fellowship from the Conseil Régional de Haute-Normandie and the CIT-IFM Recherche.

Subjects
MESH: Signal Transduction, Time Factors, MAP Kinase Kinase 4, MESH: Drug Interactions, MESH: Neurons, Fluorescent Antibody Technique, MESH: Neurotransmitter Agents, MESH: Neuropeptides, MESH: Caspase 9, MESH: Animals, Newborn, MESH: Dose-Response Relationship, Drug, Membrane Potentials, Mice, Cytosol, MESH: Cytosol, Sphingosine, MESH: Reverse Transcriptase Polymerase Chain Reaction, Cerebellum, MESH: Caspase 3, MESH: Animals, Drug Interactions, Enzyme Inhibitors, MESH: Fluorescent Antibody Technique, Cells, Cultured, bcl-2-Associated X Protein, Neurons, Neurotransmitter Agents, Caspase 3, Reverse Transcriptase Polymerase Chain Reaction, Intracellular Signaling Peptides and Proteins, Cytochromes c, MESH: Cytochromes c, Carbocyanines, MESH: Gene Expression Regulation, Caspase 9, Mitochondria, ERK, Proto-Oncogene Proteins c-bcl-2, MESH: Enzyme Inhibitors, [SDV.TOX]Life Sciences [q-bio]/Toxicology, Caspases, Pituitary Adenylate Cyclase-Activating Polypeptide, MESH: Carbocyanines, MESH: Sphingosine, Mitogen-Activated Protein Kinases, hormones, hormone substitutes, and hormone antagonists, MESH: MAP Kinase Kinase 4, MESH: Cells, Cultured, Signal Transduction, endocrine system, MESH: Rats, MESH: Mitochondria, Blotting, Western, Models, Neurological, MESH: Mitogen-Activated Protein Kinase Kinases, mitochondrial apoptotic pathway, MESH: Models, Neurological, MESH: Intracellular Signaling Peptides and Proteins, MESH: Membrane Potentials, MESH: Blotting, Western, Animals, Bcl-2, MESH: bcl-2-Associated X Protein, Nerve Growth Factors, RNA, Messenger, Rats, Wistar, MESH: Mice, MESH: RNA, Messenger, Mitogen-Activated Protein Kinase Kinases, MESH: Caspases, Dose-Response Relationship, Drug, MESH: Nerve Growth Factors, MESH: Pituitary Adenylate Cyclase-Activating Polypeptide, MESH: Time Factors, Neuropeptides, JNK Mitogen-Activated Protein Kinases, MESH: JNK Mitogen-Activated Protein Kinases, MESH: Rats, Wistar, MESH: Mitogen-Activated Protein Kinases, MESH: Cerebellum, Rats, MESH: Proto-Oncogene Proteins c-bcl-2, Animals, Newborn, Gene Expression Regulation, Bax, Benzimidazoles, JNK, MESH: Benzimidazoles
Abstract

International audience; The sphingomyelin-derived messenger ceramides provoke neuronal apoptosis through caspase-3 activation, while the neuropeptide pituitary adenylate cyclase-activating polypeptide (PACAP) promotes neuronal survival and inhibits caspase-3 activity. However, the mechanisms leading to the opposite regulation of caspase-3 by C2-ceramide and PACAP are currently unknown. Here, we show that PACAP prevents C2-ceramide-induced inhibition of mitochondrial potential and C2-ceramide-evoked cytochrome c release. C2-ceramide stimulated Bax expression, but had no effect on Bcl-2, while PACAP abrogated the action of C2-ceramide on Bax and stimulated Bcl-2 expression. The effects of C2-ceramide and PACAP on Bax and Bcl-2 were blocked, respectively, by the JNK inhibitor L-JNKI1 and the MEK inhibitor U0126. L-JNKI1 prevented the alteration of mitochondria induced by C2-ceramide while U0126 suppressed the protective effect of PACAP against the deleterious action of C2-ceramide on mitochondrial potential. Moreover, L-JNKI1 inhibited the stimulatory effect of C2-ceramide on caspase-9 and -3 and prevented C2-ceramide-induced cell death. U0126 blocked PACAP-induced Bcl-2 expression, abrogated the inhibitory effect of PACAP on ceramide-induced caspase-9 activity, and promoted granule cell death. The present study reveals that C2-ceramide and PACAP exert opposite effects on Bax and Bcl-2 through, respectively, JNK- and ERK-dependent mechanisms. These data indicate that the mitochondrial pathway plays a pivotal role in the pro- and anti-apoptotic effects of C2-ceramide and PACAP.

Published
2004
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30. Pituitary adenylate cyclase-activating polypeptide (PACAP) stimulates endozepine release from cultured rat astrocytes via a PKA-dependent mechanism

Author

Christine Patte-Mensah, Pierrick Gandolfo, Hubert Vaudry, Olfa Masmoudi, Marie-Christine Tonon, Jérôme Leprince, David Vaudry, Alain Fournier, Neuroendocrinologie cellulaire et moléculaire, Université de Rouen Normandie (UNIROUEN), Normandie Université (NU)-Normandie Université (NU)-Institut National de la Santé et de la Recherche Médicale (INSERM), Institut Fédératif de Recherches Multidisciplinaires sur les Peptides (IFRMP 23), Institut National de la Santé et de la Recherche Médicale (INSERM)-Centre de Lutte Contre le Cancer Henri Becquerel Normandie Rouen (CLCC Henri Becquerel)-Université de Rouen Normandie (UNIROUEN), Normandie Université (NU)-Normandie Université (NU)-Université Le Havre Normandie (ULH), Normandie Université (NU)-CHU Rouen, Normandie Université (NU)-Centre National de la Recherche Scientifique (CNRS), Institut Armand Frappier (INRS-IAF), and Institut National de la Recherche Scientifique [Québec] (INRS)-Réseau International des Instituts Pasteur (RIIP)

Subjects
MESH: Signal Transduction, [SDV.NEU.NB]Life Sciences [q-bio]/Neurons and Cognition [q-bio.NC]/Neurobiology, [CHIM.THER]Chemical Sciences/Medicinal Chemistry, MESH: Neuropeptides, Biochemistry, MESH: Dose-Response Relationship, Drug, chemistry.chemical_compound, 0302 clinical medicine, Phosphatidylinositol Phosphates, [SDV.BC.IC]Life Sciences [q-bio]/Cellular Biology/Cell Behavior [q-bio.CB], Cyclic AMP, MESH: Animals, MESH: Peptide Fragments, Cells, Cultured, MESH: Cyclic AMP, Diazepam Binding Inhibitor, 0303 health sciences, MESH: Kinetics, hemic and immune systems, respiratory system, MESH: Phosphatidylinositol Phosphates, 3. Good health, MESH: Calcium, Pituitary Adenylate Cyclase-Activating Polypeptide, hormones, hormone substitutes, and hormone antagonists, Signal Transduction, Biotechnology, MESH: Cells, Cultured, medicine.medical_specialty, endocrine system, Endozepine, MESH: Rats, Neuropeptide, Adenylate kinase, MESH: Cyclic AMP-Dependent Protein Kinases, Biology, Cyclase, 03 medical and health sciences, Internal medicine, Genetics, medicine, Animals, Secretion, Molecular Biology, Protein kinase C, 030304 developmental biology, Dose-Response Relationship, Drug, Neuropeptides, MESH: Pituitary Adenylate Cyclase-Activating Polypeptide, Antagonist, Cyclic AMP-Dependent Protein Kinases, Peptide Fragments, Rats, MESH: Astrocytes, Kinetics, Endocrinology, Chelerythrine, chemistry, Astrocytes, [SDV.SP.PHARMA]Life Sciences [q-bio]/Pharmaceutical sciences/Pharmacology, MESH: Diazepam Binding Inhibitor, Calcium, 030217 neurology & neurosurgery
Abstract

International audience; Astroglial cells synthesize and release endozepines, neuropeptides that are related to the octadecaneuropeptide ODN. Glial cells also express PACAP/VIP receptors. We have investigated the possible effect of PACAP on the release of ODN-like immunoreactivity (ODN-LI) by cultured rat astrocytes. Administration of PACAP27 and PACAP38 induced a concentration-dependent increase in secretion of ODN-LI whereas VIP was approximately 1000-fold less potent. The maximum effect of PACAP38 occurred after 5 min, then gradually declined during the next 10 min. The stimulatory effects of PACAP and VIP were abrogated by the PACAP antagonist PACAP6-38. PACAP38 stimulated cAMP formation, activated polyphosphoinositide turnover, and provoked calcium mobilization from IP3-sensitive pools. The PKA inhibitor H89 suppressed PACAP-induced secretion of ODN-LI, whereas PLC inhibitor U73122 and the PKC inhibitor chelerythrine had no effect. In contrast, U73122 restored the stimulatory action of PACAP on ODN-LI release and cAMP formation during prolonged (15 min) incubation with the peptide, and this effect was prevented by PMA. The present results demonstrate that PACAP stimulates endozepine release through activation of PAC1 receptors coupled to the AC/PKA pathway. Our data also show that activation of the PLC/PKC pathway down-regulates the effect of PACAP on endozepine release.

Published
2003
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31. Pituitary growth hormone secretion in the turbot, a phylogenetically recent teleost, is regulated by a species-specific pattern of neuropeptides

Author

Nadine Le Belle, Gilles Boeuf, Billy K. C. Chow, J. Marchelidon, Karine Rousseau, Sylvie Dufour, Karine Pichavant, Biologie des organismes marins et écosystèmes (BOME), Centre National de la Recherche Scientifique (CNRS)-Université Pierre et Marie Curie - Paris 6 (UPMC)-Muséum national d'Histoire naturelle (MNHN), Optimisation des régulations physiologiques (ORPHY (EA 4324)), Institut Brestois Santé Agro Matière (IBSAM), Université de Brest (UBO)-Université de Brest (UBO)-Université de Brest (UBO)-Centre Hospitalier Régional Universitaire de Brest (CHRU Brest), Littoral, Environnement, Télédétection, Géomatique (LETG - Rennes), Littoral, Environnement, Télédétection, Géomatique UMR 6554 (LETG), Université de Caen Normandie (UNICAEN), Normandie Université (NU)-Normandie Université (NU)-Université d'Angers (UA)-École pratique des hautes études (EPHE), Université Paris sciences et lettres (PSL)-Université Paris sciences et lettres (PSL)-Université de Brest (UBO)-Université de Rennes 2 (UR2), Université de Rennes (UNIV-RENNES)-Université de Rennes (UNIV-RENNES)-Centre National de la Recherche Scientifique (CNRS)-Institut de Géographie et d'Aménagement Régional de l'Université de Nantes (IGARUN), Université de Nantes (UN)-Université de Nantes (UN)-Université de Caen Normandie (UNICAEN), Université de Nantes (UN)-Université de Nantes (UN), Muséum national d'Histoire naturelle (MNHN)-Université Pierre et Marie Curie - Paris 6 (UPMC)-Centre National de la Recherche Scientifique (CNRS), Université de Brest (UBO)-Centre Hospitalier Régional Universitaire de Brest (CHRU Brest)-Institut Brestois Santé Agro Matière (IBSAM), Université de Brest (UBO)-Université de Brest (UBO), Normandie Université (NU)-Normandie Université (NU)-Université d'Angers (UA)-École Pratique des Hautes Études (EPHE), Université Paris sciences et lettres (PSL)-Université Paris sciences et lettres (PSL)-Université de Brest (UBO)-Université de Rennes 2 (UR2)-Centre National de la Recherche Scientifique (CNRS)-Institut de Géographie et d'Aménagement Régional de l'Université de Nantes (IGARUN), and Guerrero, François

Subjects
MESH: Signal Transduction, MESH: Eels, [SDV.MHEP.PHY] Life Sciences [q-bio]/Human health and pathology/Tissues and Organs [q-bio.TO], Endocrinology, Diabetes and Metabolism, MESH: Somatostatin, MESH: Neuropeptides, Culture Media, Serum-Free, MESH: Drug Synergism, MESH: Dose-Response Relationship, Drug, chemistry.chemical_compound, 0302 clinical medicine, Endocrinology, MESH: Pituitary Gland, MESH: Animals, Cells, Cultured, 0303 health sciences, Eels, Forskolin, MESH: Pituitary Gland, Anterior, Drug Synergism, Growth hormone–releasing hormone, MESH: Culture Media, Serum-Free, MESH: Flatfishes, Somatostatin, Pituitary Gland, Flatfishes, Pituitary Adenylate Cyclase-Activating Polypeptide, Tetradecanoylphorbol Acetate, MESH: Forskolin, hormones, hormone substitutes, and hormone antagonists, Signal Transduction, MESH: Cells, Cultured, medicine.medical_specialty, endocrine system, Somatotropic cell, Growth-hormone-releasing hormone receptor, 030209 endocrinology & metabolism, In Vitro Techniques, Gonadotropic cell, 03 medical and health sciences, Cellular and Molecular Neuroscience, Species Specificity, Pituitary Gland, Anterior, Thyrotropin-releasing hormone receptor, Internal medicine, medicine, [SDV.MHEP.PHY]Life Sciences [q-bio]/Human health and pathology/Tissues and Organs [q-bio.TO], Animals, MESH: Species Specificity, MESH: Tetradecanoylphorbol Acetate, 030304 developmental biology, Dose-Response Relationship, Drug, Endocrine and Autonomic Systems, Colforsin, Neuropeptides, MESH: Pituitary Adenylate Cyclase-Activating Polypeptide, chemistry, Growth Hormone, MESH: Growth Hormone, Corticotropic cell
Abstract

In mammals, growth hormone (GH) is under a dual hypothalamic control exerted by growth hormone-releasing hormone (GHRH) and somatostatin (SRIH). We investigated GH release in a pleuronectiform teleost, the turbot (Psetta maxima), using a serum-free primary culture of dispersed pituitary cells. Cells released GH for up to 12 days in culture, indicating that turbot somatotropes do not require releasing hormone for their regulation. SRIH dose-dependently inhibited GH release up to a maximal inhibitory effect of 95%. None of the potential stimulators tested induced any change in basal GH release. Also, neither forskolin, an activator of adenylate cyclase, nor phorbol ester (TPA), an activator of protein kinase C, were able to modify GH release, suggesting that spontaneous basal release already represents the maximal secretory capacity of turbot somatotropes. In contrast, forskolin and TPA were able to increase GH release in the presence of SRIH. In this condition (coincubation with SRIH), pituitary adenylate cyclase-activating polypeptide (PACAP) stimulated GH release, whereas none of the other neuropeptides tested (GHRHs; sea bream or salmon or chicken II GnRHs; TRH; CRH) had any significant effect. These data indicate that inhibitory control by SRIH may be the basic control of GH production in teleosts and lower vertebrates, while PACAP may represent the ancestral growth hormone-releasing factor in teleosts, a role taken over in higher vertebrates by GHRH.

Published
2001

32. Strategies to Convert PACAP from a Hypophysiotropic Neurohormone Into a Neuroprotective Drug

Author

Ngoc-Duc Doan, Jérôme Leprince, Olivier Wurtz, Hubert Vaudry, David Chatenet, David Vaudry, Alain Fournier, Steve Bourgault, Institut Armand Frappier (INRS-IAF), Institut National de la Recherche Scientifique [Québec] (INRS)-Réseau International des Instituts Pasteur (RIIP), Différenciation et communication neuronale et neuroendocrine (DC2N), Université de Rouen Normandie (UNIROUEN), Normandie Université (NU)-Normandie Université (NU)-Institut National de la Santé et de la Recherche Médicale (INSERM), International Associated laboratory Samuel de Champlain, Institut National de la Santé et de la Recherche Médicale (INSERM)-Institut National de la Recherche Scientifique [Québec] (INRS)-Université de Rouen Normandie (UNIROUEN), Normandie Université (NU)-Normandie Université (NU), Plate-Forme de Recherche en Imagerie Cellulaire de Haute-Normandie (PRIMACEN), Normandie Université (NU)-Normandie Université (NU)-Institute for Research and Innovation in Biomedicine (IRIB), Normandie Université (NU)-Normandie Université (NU)-Centre National de la Recherche Scientifique (CNRS)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Université de Rouen Normandie (UNIROUEN), and Normandie Université (NU)-Centre National de la Recherche Scientifique (CNRS)-Institut National de la Santé et de la Recherche Médicale (INSERM)

Subjects
[SDV.NEU.NB]Life Sciences [q-bio]/Neurons and Cognition [q-bio.NC]/Neurobiology, Receptors, Pituitary Adenylate Cyclase-Activating Polypeptide, Apoptosis, MESH: Receptors, G-Protein-Coupled, Pharmacology, Brain Ischemia, Receptors, G-Protein-Coupled, 0302 clinical medicine, [SDV.BC.IC]Life Sciences [q-bio]/Cellular Biology/Cell Behavior [q-bio.CB], Drug Discovery, MESH: Animals, media_common, 0303 health sciences, MESH: Cytokines, Microglia, MESH: Brain Ischemia, MESH: Neuroprotective Agents, 3. Good health, medicine.anatomical_structure, Neuroprotective Agents, MESH: Cell Survival, Cytokines, Pituitary Adenylate Cyclase-Activating Polypeptide, medicine.symptom, Intracellular, hormones, hormone substitutes, and hormone antagonists, Drug, endocrine system, Cell Survival, media_common.quotation_subject, Inflammation, Biology, Neuroprotection, 03 medical and health sciences, MESH: Drug Discovery, In vivo, medicine, Animals, Humans, Nerve Growth Factors, Neuroinflammation, 030304 developmental biology, MESH: Receptors, Pituitary Adenylate Cyclase-Activating Polypeptide, MESH: Humans, MESH: Nerve Growth Factors, MESH: Apoptosis, MESH: Pituitary Adenylate Cyclase-Activating Polypeptide, MESH: Astrocytes, Astrocytes, Brain Injuries, MESH: Brain Injuries, [SDV.SP.PHARMA]Life Sciences [q-bio]/Pharmaceutical sciences/Pharmacology, 030217 neurology & neurosurgery
Abstract

International audience; In neurological insults, such as cerebral ischemia and traumatic brain injury, complex molecular mechanisms involving inflammation and apoptosis are known to cause severe neuronal cell loss, emphasizing the necessity of developing therapeutic strategies targeting simultaneously these two processes. Over the last decade, numerous in vitro and in vivo studies have demonstrated the unique therapeutical potential of pituitary adenylate cyclase-activating polypeptide (PACAP) for the treatment of neuronal disorders involving apoptotic cell death and neuroinflammation. The neuroprotective activity of PACAP is based on its capacity to reduce the production of deleterious cytokines from activated microglia, to stimulate the release of neuroprotective agents from astrocytes and to inhibit pro-apoptotic intracellular pathways. However, the use of PACAP as a clinically applicable drug is hindered by its peptidic nature. As most natural peptides, native PACAP shows poor metabolic stability, low bioavailability, inadequate distribution and rapid blood clearance. Moreover, injection of PACAP to human can induce peripheral adverse side effects. Therefore, targeted chemical modifications and/or conjugation of PACAP to different macromolecules are required to improve the pharmacokinetic and pharmacological properties of PACAP. This review presents the chemical, biochemical and pharmacological strategies that are currently under development to convert PACAP from a hypophysiotropic neurohormone into a clinically relevant neuroprotective drug.

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