29 results on '"Daniela Necchi"'
Search Results
2. CBP and p300 acetylate PCNA to link its degradation with nucleotide excision repair synthesis
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Daniela Necchi, Micol Tillhon, Sabrina Sommatis, Tiziana Nardo, Angela Bachi, Ornella Cazzalini, Ennio Prosperi, Lucia Anna Stivala, Ilaria Dutto, Alexander Rapp, M. Cristina Cardoso, and A. Ivana Scovassi
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DNA Replication ,DNA Repair ,DNA repair ,DNA-Directed DNA Polymerase ,Genome Integrity, Repair and Replication ,DNA polymerase delta ,RFC2 ,03 medical and health sciences ,0302 clinical medicine ,Replication factor C ,Proliferating Cell Nuclear Antigen ,Genetics ,Humans ,p300-CBP Transcription Factors ,Cells, Cultured ,030304 developmental biology ,0303 health sciences ,DNA clamp ,biology ,Acetylation ,DNA ,Molecular biology ,CREB-Binding Protein ,Chromatin ,Proliferating cell nuclear antigen ,030220 oncology & carcinogenesis ,Mutation ,biology.protein ,DNA mismatch repair ,Nucleotide excision repair ,DNA Damage - Abstract
The proliferating cell nuclear antigen (PCNA) protein serves as a molecular platform recruiting and coordinating the activity of factors involved in multiple deoxyribonucleic acid (DNA) transactions. To avoid dangerous genome instability, it is necessary to prevent excessive retention of PCNA on chromatin. Although PCNA functions during DNA replication appear to be regulated by different post-translational modifications, the mechanism regulating PCNA removal and degradation after nucleotide excision repair (NER) is unknown. Here we report that CREB-binding protein (CBP), and less efficiently p300, acetylated PCNA at lysine (Lys) residues Lys13,14,77 and 80, to promote removal of chromatin-bound PCNA and its degradation during NER. Mutation of these residues resulted in impaired DNA replication and repair, enhanced the sensitivity to ultraviolet radiation, and prevented proteolytic degradation of PCNA after DNA damage. Depletion of both CBP and p300, or failure to load PCNA on DNA in NER deficient cells, prevented PCNA acetylation and degradation, while proteasome inhibition resulted in accumulation of acetylated PCNA. These results define a CBP and p300-dependent mechanism for PCNA acetylation after DNA damage, linking DNA repair synthesis with removal of chromatin-bound PCNA and its degradation, to ensure genome stability.
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- 2014
3. Nature-Inspired Multifunctional Ligands: Focusing on Amyloid-Based Molecular Mechanisms of Alzheimer’s Disease
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Jessica Fiori, Cristina Lanni, Elena Simoni, Manuela Bartolini, Michela Rosini, Roberta Caporaso, Daniela Necchi, Anna Minarini, Antonella Pinto, Vincenza Andrisano, Melania Maria Serafini, Simoni, Elena, Serafini, Melania M., Bartolini, Manuela, Caporaso, Roberta, Pinto, Antonella, Necchi, Daniela, Fiori, Jessica, Vincenza, Andrisano, Minarini, Anna, Lanni, Cristina, and Rosini, Michela
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0301 basic medicine ,Amyloid ,Cell Survival ,Peptide ,Protein Serine-Threonine Kinases ,medicine.disease_cause ,Ligands ,Protective Agents ,Biochemistry ,03 medical and health sciences ,Mediator ,Alzheimer Disease ,Cell Line, Tumor ,Drug Discovery ,medicine ,Humans ,General Pharmacology, Toxicology and Pharmaceutics ,Pharmacology ,chemistry.chemical_classification ,Reactive oxygen species ,Amyloid beta-Peptides ,Organic Chemistry ,Hydrogen Peroxide ,medicine.disease ,Zyxin ,Oxidative Stress ,030104 developmental biology ,chemistry ,Cinnamates ,Molecular Medicine ,Alzheimer’s disease · antioxidants · amyloid-beta peptide · p53 · multifunctional ligands ,Signal transduction ,Alzheimer's disease ,Tumor Suppressor Protein p53 ,Carrier Proteins ,Reactive Oxygen Species ,Oxidative stress ,Function (biology) ,Signal Transduction - Abstract
The amyloidogenic pathway is a prominent feature of Alzheimer's disease (AD). However, growing evidence suggests that a linear disease model based on β-amyloid peptide (Aβ) alone is not likely to be realistic, which therefore calls for further investigations on the other actors involved in the play. The pro-oxidant environment induced by Aβ in AD pathology is well established, and a correlation among Aβ, oxidative stress, and conformational changes in p53 has been suggested. In this study, we applied a multifunctional approach to identify allyl thioesters of variously substituted trans-cinnamic acids for which the pharmacological profile was strategically tuned by hydroxy substituents on the aromatic moiety. Indeed, only catechol derivative 3 [(S)-allyl (E)-3-(3,4-dihydroxyphenyl)prop-2-enethioate] inhibited Aβ fibrilization. Conversely, albeit to different extents, all compounds were able to decrease the formation of reactive oxygen species in SH-SY5Y neuroblastoma cells and to prevent alterations in the conformation of p53 and its activity mediated by soluble sub-lethal concentrations of Aβ. This may support an involvement of oxidative stress in Aβ function, with p53 emerging as a potential mediator of their functional interplay.
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- 2016
4. p300/CBP acetyl transferases interact with and acetylate the nucleotide excision repair factor XPG
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Tiziana Nardo, Ornella Cazzalini, Lucia Anna Stivala, Micol Tillhon, Daniela Necchi, Sabrina Sommatis, Ennio Prosperi, and A. Ivana Scovassi
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Cyclin-Dependent Kinase Inhibitor p21 ,Curcumin ,DNA Repair ,DNA repair ,DNA damage ,DNA Mismatch Repair ,Biochemistry ,chemistry.chemical_compound ,Proliferating Cell Nuclear Antigen ,Humans ,p300-CBP Transcription Factors ,RNA, Small Interfering ,Molecular Biology ,biology ,DNA replication ,Nuclear Proteins ,Acetylation ,Cell Biology ,Histone acetyltransferase ,Fibroblasts ,Endonucleases ,Molecular biology ,Proliferating cell nuclear antigen ,DNA-Binding Proteins ,chemistry ,biology.protein ,DNA mismatch repair ,DNA ,HeLa Cells ,Transcription Factors ,Nucleotide excision repair - Abstract
Nucleotide excision repair (NER) is an important DNA repair mechanism through which cells remove bulky DNA lesions. Following DNA damage, the histone acetyltransferase (HAT) p300 (also referred to as lysine acetyltransferase or KAT) is known to associate with proliferating cell nuclear antigen (PCNA), a master regulator of DNA replication and repair processes. This interaction, which results in HAT inhibition, may be dissociated by the cell cycle inhibitor p21(CDKN1A), thereby restoring p300 activity; however, the role of this protein interplay is still unclear. Here, we report that silencing p300 or its homolog CREB-binding protein (CBP) by RNA interference (RNAi) significantly reduces DNA repair synthesis in human fibroblasts. In addition, we determined whether p300 and CBP may associate with and acetylate specific NER factors such as XPG, the 3'-endonuclease that is involved in the incision/excision step and is known to interact with PCNA. Our results show that p300 and CBP interact with XPG, which has been found to be acetylated in vivo. XPG is acetylated by p300 in vitro, and this reaction is inhibited by PCNA. Knocking down both p300/CBP by RNAi or by chemical inhibition with curcumin greatly reduced XPG acetylation, and a concomitant accumulation of the protein at DNA damage sites was observed. The ability of p21 to bind PCNA was found to regulate the interaction between p300 and XPG, and an abnormal accumulation of XPG at DNA damage sites was also found in p21(-/-) fibroblasts. These results indicate an additional function of p300/CBP in NER through the acetylation of XPG protein in a PCNA-p21 dependent manner.
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- 2012
5. Dysfunction of the ubiquitin–proteasome system in the cerebellum of aging Ts65Dn mice
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Elda Scherini, Selene Lomoio, and Daniela Necchi
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Aging ,Proteasome Endopeptidase Complex ,Cerebellum ,Immunocytochemistry ,Purkinje cell ,Biology ,Mice ,Mice, Neurologic Mutants ,Purkinje Cells ,Developmental Neuroscience ,Western blot ,Ubiquitin ,mental disorders ,medicine ,Animals ,Cell Nucleus ,medicine.diagnostic_test ,Neurodegeneration ,Ubiquitination ,medicine.disease ,Molecular biology ,Disease Models, Animal ,medicine.anatomical_structure ,nervous system ,Neurology ,Proteasome ,Nerve Degeneration ,biology.protein ,Neuron ,Down Syndrome - Abstract
In the cerebellum of adult-aging Ts65Dn mice, a murine model of Down syndrome, Purkinje cells undergo degeneration. Searching for the cause of Purkinje cell degeneration, we have studied the ubiquitin–proteasome system (UPS) in the cerebellum of aging Ts65Dn mice. Inhibition of UPS is sufficient to induce neuron degeneration and death. Proteasome chymotrypsin-like proteolytic activity was reduced by 35% in the cerebellum of Ts65Dn mice in comparison with euploid animals. Accordingly, Western blot analysis of ubiquitin showed an increase in ubiquitinated proteins. Immunocytochemistry for ubiquitin revealed strongly positive intranuclear inclusions in Purkinje cells and large neurons of cerebellar nuclei. The Western blot analysis of ubiquitin in nuclear protein extracts confirmed the increase of ubiquitinated proteins in the cell nuclei. After FUS immunocytochemistry, large intranuclear inclusions were visible in Purkinje cells and large neurons of cerebellar nuclei in Ts65Dn mice. Together, data indicate a possible role for proteasome inhibition in the cerebellar neurodegeneration in Ts65Dn mice.
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- 2011
6. β-Amyloid overload does not directly correlate with SAPK/JNK activation and tau protein phosphorylation in the cerebellar cortex of Ts65Dn mice
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Daniela Necchi, Selene Lomoio, and Elda Scherini
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Male ,Cerebellum ,Tau protein ,Purkinje cell ,Mice, Transgenic ,Trisomy ,tau Proteins ,Axon hillock ,Mice ,Purkinje Cells ,Interneurons ,mental disorders ,medicine ,Amyloid precursor protein ,Animals ,Genetic Predisposition to Disease ,Mitogen-Activated Protein Kinase 8 ,Phosphorylation ,Molecular Biology ,Amyloid beta-Peptides ,biology ,General Neuroscience ,JNK Mitogen-Activated Protein Kinases ,Granule cell ,Axons ,Cell biology ,Disease Models, Animal ,medicine.anatomical_structure ,Tauopathies ,nervous system ,Cerebellar cortex ,Nerve Degeneration ,biology.protein ,Female ,Neurology (clinical) ,Neuron ,Down Syndrome ,Neuroglia ,Neuroscience ,Signal Transduction ,Developmental Biology - Abstract
It is known that in the nervous tissue beta-amyloid overproduction and its extracellular or intracellular deposition can activate mitogen-activated protein kinases involved in tau protein phosphorylation. Hyperphosphorylated tau is not more able to bind neuron microtubules, leading to their disassembly and axon degeneration. We have previously described that at 10 months of age in the cerebellum of Ts65Dn mice, which are partially trisomic for the chromosome 16 and are considered a valuable model for Down syndrome, Purkinje cells undergo axon degeneration. Taking into consideration that Ts65Dn mice carry three copies of the gene encoding for the amyloid precursor protein, to characterize potential signaling events triggering the degenerative phenomenon, specific antibodies were used to examine the role of beta-amyloid overload in the activation of the stress activated kinase/c-jun N-terminal kinase (SAPK/JNK) and tau protein phosphorylation in the cerebellar cortex of 12-month-old Ts65Dn mice. We found small extracellular deposits of beta-amyloid at the borderline between the granule cell layer and the white matter, i.e., in the vicinity of the area where calbindin immunostaining of Purkinje cell axons revealed clusters of newly formed terminals of injured axons. Moreover, intracellular deposits were present in the somata of Purkinje cells. The level of activation of SAPK/JNK was greatly increased. The activation occurred in the "pinceaux" made by basket interneuron axons at the axon hillock of Purkinje cells. Antibody directed against tau protein phosphorylated at Ser-396/Ser-404 revealed positive NG2 cells and Bergman fibers in the molecular layer and oligodendrocytes in the white matter. Data indicate that beta-amyloid extracellular deposits could have exerted a local cytotoxic effect, leading to Purkinje cell axon degeneration. The activation of SAPK/JNK in basket cell "pinceaux" may be a consequence of altered functionality of Purkinje cells and may represent an attempt of basket cells of synaptic remodeling. Moreover, the findings for tau protein phosphorylation suggest that Ts65Dn mice are affected by a cerebellar glial tauopathy.
- Published
- 2009
7. Altered Neuron Excitability and Synaptic Plasticity in the Cerebellar Granular Layer of Juvenile Prion Protein Knock-Out Mice with Impaired Motor Control
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Francesca Prestori, Bertrand Bearzatto, Jeanne Lainé, Egidio D'Angelo, Daniela Necchi, Serge N. Schiffmann, Shyam Diwakar, Herbert Axelrad, and Paola Rossi
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Cerebellum ,Patch-Clamp Techniques ,Prions ,Movement ,animal diseases ,Stimulation ,Biology ,Membrane Potentials ,Mice ,Microscopy, Electron, Transmission ,Neural Pathways ,Neuroplasticity ,medicine ,Animals ,Cell Proliferation ,Mice, Knockout ,Neurons ,Movement Disorders ,Neuronal Plasticity ,Transmissible spongiform encephalopathy ,General Neuroscience ,Age Factors ,Excitatory Postsynaptic Potentials ,Dose-Response Relationship, Radiation ,Long-term potentiation ,Articles ,medicine.disease ,Granule cell ,Electric Stimulation ,nervous system diseases ,Mice, Inbred C57BL ,medicine.anatomical_structure ,Animals, Newborn ,Bromodeoxyuridine ,Synaptic plasticity ,Neuron ,Neuroscience - Abstract
Although the role of abnormal prion protein (PrP) conformation in generating infectious brain diseases (transmissible spongiform encephalopathy) has been recognized, the function of PrP in the normal brain remains mostly unknown. In this investigation, we considered the effect of PrP gene knock-out (PrP0/0) on cerebellar neural circuits and in particular on granule cells, which show intense PrP expression during development and selective affinity for PrP. At the third postnatal week, when PrP expression would normally attain mature levels, PrP0/0mice showed low performance in the accelerating rotarod and runway tests and the functioning of 40% of granule cells was abnormal. Spikes were slow, nonovershooting, and nonrepetitive in relation with a reduction in transient inward and outward membrane currents, and also the EPSPs and EPSCs had slow kinetics. Overall, these alterations closely resembled an immature phenotype. Moreover, in slow-spiking PrP0/0granule cells, theta-burst stimulation was unable to induce any long-term potentiation. This profound impairment in synaptic excitation and plasticity was associated with a protracted proliferation of granule cells and disappeared at P40–P50 along with the recovery of normal motor behavior (Büeler et al., 1992). These results suggest that PrP plays an important role in granule cell development eventually regulating cerebellar network formation and motor control.
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- 2008
8. Spatiotemporal dynamics of p21CDKN1A protein recruitment to DNA-damage sites and interaction with proliferating cell nuclear antigen
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Oliver Mortusewicz, Daniela Necchi, Monica Savio, Lucia Anna Stivala, Heinrich Leonhardt, Tiziana Nardo, M. Cristina Cardoso, Ennio Prosperi, Ornella Cazzalini, and Paola Perucca
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Cyclin-Dependent Kinase Inhibitor p21 ,DNA Repair ,Ultraviolet Rays ,DNA damage ,DNA repair ,Transfection ,DNA polymerase delta ,Cell Line ,Green fluorescent protein ,Mice ,Mutant protein ,Proliferating Cell Nuclear Antigen ,Animals ,Humans ,biology ,Cell Cycle ,DNA replication ,Cell Biology ,Molecular biology ,Chromatin ,Proliferating cell nuclear antigen ,Cell biology ,DNA-Binding Proteins ,biology.protein ,DNA Damage ,HeLa Cells ,Protein Binding ,Transcription Factors ,Nucleotide excision repair - Abstract
The cyclin-dependent kinase inhibitor p21CDKN1A plays a fundamental role in the DNA-damage response by inducing cell-cycle arrest, and by inhibiting DNA replication through association with the proliferating cell nuclear antigen (PCNA). However, the role of such an interaction in DNA repair is poorly understood and controversial. Here, we provide evidence that a pool of p21 protein is rapidly recruited to UV-induced DNA-damage sites, where it colocalises with PCNA and PCNA-interacting proteins involved in nucleotide excision repair (NER), such as DNA polymerase δ, XPG and CAF-1. In vivo imaging and confocal fluorescence microscopy analysis of cells coexpressing p21 and PCNA fused to green or red fluorescent protein (p21-GFP, RFP-PCNA), showed a rapid relocation of both proteins at microirradiated nuclear spots, although dynamic measurements suggested that p21-GFP was recruited with slower kinetics. An exogenously expressed p21 mutant protein unable to bind PCNA neither colocalised, nor coimmunoprecipitated with PCNA after UV irradiation. In NER-deficient XP-A fibroblasts, p21 relocation was greatly delayed, concomitantly with that of PCNA. These results indicate that early recruitment of p21 protein to DNA-damage sites is a NER-related process dependent on interaction with PCNA, thus suggesting a direct involvement of p21 in DNA repair.
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- 2006
9. The malformation of the cerebellar fissura prima : a tool for studying histogenetic processes
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Daniela Necchi and Elda Scherini
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Models, Anatomic ,Cell type ,Cerebellum ,Pia mater ,Incidence ,Meninges ,Histogenesis ,Biology ,Nervous System Malformations ,Rats ,medicine.anatomical_structure ,Species Specificity ,Neurology ,Cytoarchitecture ,Cerebellar cortex ,medicine ,Animals ,Humans ,Basal lamina ,Neurology (clinical) ,Neuroscience - Abstract
Due to its regular cytoarchitecture and the relatively low number of cell types, the development of the cerebellar cortex is a model of election for studies of morphogenetic processes. To unravel the cellular and molecular mechanisms that regulate cell development, migration and differentiation and the settling of local circuits, pertubation of the three-layered organization of the cerebellar cortex has been induced by X-ray irradiation or antimitotic drug. In this review we deal with some data about the incidence and development of the malformation of the cerebellar fissura prima of the rat, as an eligible model for histogenetic studies. The naturally occurring malformation of the fissura prima is characterized by the loss of the three-layered organization of folia and the presence of large masses of ectopic granule cells. The malformation appears to be under genetic control, since the incidence of affected animals is consistent over extended breeding cycles, although the target of the eventual mutation is unknown. The observation of development of the malformation in infant rats suggests that a defect in meninges, and in particular in the pia mater, is a primary contributing factor. The molecules responsible for this defect are not identified, but they must be involved in basal lamina stabilization or destabilization. In fact, there is evidence that meninges do develop during first stages of histogenesis and only later degenerate. A possible correlation with certain human pathologies that involve defects in foliation is discussed.
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- 2002
10. Increase of the ornithine decarboxylase/polyamine system and transglutaminase upregulation in the spinal cord of aged rats
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Daniela Necchi, Elda Scherini, Antonio Contestabile, and Marco Virgili
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Male ,Aging ,Cerebellum ,medicine.medical_specialty ,Central nervous system ,Biology ,Protein polyamination ,Ornithine Decarboxylase ,Hippocampus ,Ornithine decarboxylase ,chemistry.chemical_compound ,Internal medicine ,medicine ,Animals ,Rats, Wistar ,Cerebral Cortex ,Neurons ,Transglutaminases ,General Neuroscience ,Biogenic Polyamines ,Spinal cord ,Rats ,Up-Regulation ,Spermidine ,medicine.anatomical_structure ,Endocrinology ,Spinal Cord ,chemistry ,Biochemistry ,Putrescine ,Polyamine - Abstract
We have investigated changes in ornithine decarboxylase (ODC) activity and in polyamine levels in the central nervous system of aged rats. We measured a significant increase of ODC catalytic activity in the spinal cord from 30 month-old rats (+105%) as compared to 4 month-old rats. No changes were noticed in the cerebellum, cortex and hippocampus from the same animals. A related putrescine increase was measured in the spinal cord of 30 month-old rats (+168%), together with a smaller increase of spermidine (+33%). A parallel increase (+78%) of the Ca2+-dependent transglutaminase activity was detected in the spinal cord of 30 month-old rats, while no changes were apparent in the cortex and cerebellum. Our observations indicate a possible role of the ODC/polyamine system during the normal process of ageing in rats and point to the spinal cord as the most sensitive area for this kind of modification. A possible role of protein polyamination by tranglutaminase is discussed.
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- 2001
11. Nitric oxide-containing neurons in the nervous ganglia ofHelix aspersa during rest and activity: Immunocytochemical and enzyme histochemical detection
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Carla Fenoglio, Elena Conforti, Elda Scherini, Graziella Bernocchi, Daniela Necchi, and Maria Bonaria Pisu
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Rest ,Neuropeptide ,Snail ,Motor Activity ,Nicotinamide adenine dinucleotide ,Nitric Oxide ,Nervous System ,Neuroprotection ,Nitric oxide ,chemistry.chemical_compound ,Immunolabeling ,Antibody Specificity ,biology.animal ,Animals ,Neurons ,Behavior, Animal ,biology ,Helix, Snails ,General Neuroscience ,NADPH Dehydrogenase ,Colocalization ,Immunohistochemistry ,Molecular biology ,Ganglia, Invertebrate ,Nitric oxide synthase ,chemistry ,Biochemistry ,biology.protein ,Nitric Oxide Synthase - Abstract
Nitric oxide synthase (NOS) immunoreactivity and staining for nicotinamide adenine dinucleotide phosphate-diaphorase (NADPH-diaphorase) activity are two cytochemical markers for nitric oxide (NO)-containing neurons. The authors examined the changes in the distribution of NOS immunolabeling and NADPH-diaphorase reactivity in the cerebral and buccal ganglia of the terrestrial snail Helix aspersa during resting and active phases. During inactivity and after 1 day of activity, in the mesocerebrum and metacerebrum of the snails, there were several reactive neurons for both markers; after 7 days of activity, the number of reactive neurons was lower. Opposite results were obtained in the buccal ganglia, in which increased staining and numbers of reactive neurons were present in the active snails (after 1 day and 7 days of activity). Although the staining patterns for the two reactions were similar, colocalization was not always observed. The comparison between inactive and active animals provided a more precise survey of NOS-containing neurons in the snail cerebral ganglia than previously described. Moreover, it suggested that not only is NO involved in distinct nervous circuits, but, as a ubiquitous molecule, it also plays a role in neuroprotection and neuropeptide release. J. Comp. Neurol. 409:274–284, 1999. © 1999 Wiley-Liss, Inc.
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- 1999
12. Distribution of calretinin-like immunoreactivity in the brain of Rana esculenta
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Graziella Bernocchi, Daniela Necchi, Maria Bonaria Pisu, Cristiana Soldani, and Elda Scherini
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Brain Chemistry ,Male ,Blotting, Western ,Immunocytochemistry ,Thalamus ,Rana esculenta ,Nerve Tissue Proteins ,Sensory system ,Anatomy ,Biology ,Immunohistochemistry ,Rats ,Rana ,Cellular and Molecular Neuroscience ,Immunolabeling ,S100 Calcium Binding Protein G ,Habenula ,nervous system ,Calbindin 2 ,Tegmentum ,Animals ,Humans ,Calretinin - Abstract
The distribution of calretinin-like immunoreactivity has been analyzed in the brain of Rana esculenta. Several neurons of nuclei belonging to sensory pathways, subhabenular area and left habenula were immunopositive. Immunoreactivity was present in fibers of motor and sensory pathways, thalamus, tegmentum and isthmus. The immunolabeling pattern partially overlapped that previously described in the rat. However, in comparison with the rat, fewer cells and fibers were immunoreactive and there were less positive brain nuclei. especially in the pallium, septum and striatum, that were totally negative. Taking into consideration that these regions are rather simple in the frog, the presence of calretinin seems to be consistent with the degree of complexity of brain areas and segregation of different nuclei.
- Published
- 1999
13. Bioactive peptides and serotonin immunocytochemistry in the cerebral ganglia of hibernatingHelix aspersa
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Elda Scherini, Graziella Bernocchi, C. Vignola, Daniela Necchi, and Maria Bonaria Pisu
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Hibernation ,medicine.medical_specialty ,Immunocytochemistry ,Substance P ,General Medicine ,Neurotransmission ,Biology ,Calcitonin gene-related peptide ,chemistry.chemical_compound ,Endocrinology ,medicine.anatomical_structure ,nervous system ,chemistry ,Internal medicine ,Neuromodulation ,medicine ,Animal Science and Zoology ,Serotonin ,Neuron - Abstract
The role of some neuromodulators and neurotransmitters in the functioning of molluskan cerebral neurons and in their metabolic changes during hibernation has been considered. The cerebral ganglion of mollusks is a center for the integration of different inputs from the sensory areas of the head and for the generation of motor command impulses. During hibernation, animals are deprived of many external sensory stimuli and do not have locomotion and feeding. Immunocytochemistry for bioactive peptides (BAPs), such as SP (Substance P), CCK8 (Cholecystokinin 8/Gastrin), CGRP (Calcitonin-Gene-Related Peptide) and ET (Endothelin), and serotonin was performed on cerebral ganglia of active and hibernating Helix aspersa. The distribution of the immunopositivity was analyzed in different cell-containing areas (procerebrum, mesocerebrum, metacerebrum) and in the neuropiles. With all the antibodies raised against peptides, we observed that only a few neurons, mainly of small and medium size, had immunopositivity during the period of activity, the patterns of distribution being quite similar to those previously described in Helix or other gastropods. Fibers and varicosities with BAP immunopositivity were found in the procerebral and central neuropiles and sometimes around neurons. Serotonin-immunopositive neurons, including the giant neuron, were observed in the metacerebrum; numerous fibers and varicosities immunopositive for serotonin were present in the neuropile areas. In hibernating snails, the number of fibers with BAP and serotonin immunopositivity decreased in several areas of the neuropiles. Moreover, an increased number of neurons of the metacerebrum (two- to four-fold) and mesocerebrum (8- to 28-fold) had BAP-like immunopositivity, and the intensity of the immunoreaction for serotonin of the metacerebral neurons was also higher than in the active snails. These results are discussed, taking into account two hypotheses. The first hypothesis assumes that the increased immunocytochemical staining was really linked to accumulation of BAPs and serotonin. The second hypothesis considers that the antibodies for BAPs recognized a preprotein, the synthesis of BAPs being completed during the active period only. Both the hypotheses account for the co-occurrence and co-localization of two or ore peptides and serotonin and stress that the hibernation condition is of interest for studies on the actual function of single neurons in the cerebral ganglia. Finally, the data are consistent with the changes recently found in other markers of the morphological and functional activity of neurons, demonstrating that the neuromodulation and the neurotransmission are slowed during hibernation. J. Exp. Zool. 280:354–367, 1998. © 1998 Wiley-Liss, Inc.
- Published
- 1998
14. Small ribosomal subunits associate with nuclear myosin and actin in transit to the nuclear pores
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Barbara Cisterna, Daniela Necchi, Ennio Prosperi, and Marco Biggiogera
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Nucleolus ,Ribosome biogenesis ,macromolecular substances ,Myosins ,Biology ,Biochemistry ,Myosin ,Genetics ,ultrastructural immunocytochemistry ,Humans ,Nuclear pore ,Molecular Biology ,Cell Nucleus ,nucleocytoplasmic transport ,Ribosomal Protein S6 ,Nucleoplasm ,Eukaryotic Large Ribosomal Subunit ,ribosomal protein S6 ,Ribosomal RNA ,Flow Cytometry ,Immunohistochemistry ,Actins ,Chromatin ,Cell biology ,Microscopy, Electron ,Protein Transport ,RNA, Ribosomal ,Ribosomal protein s6 ,Nuclear Pore ,HeLa Cells ,Biotechnology - Abstract
We have followed at high resolution the ribosomal protein S6 entering the nucleus of HeLa cells, stopping in some (not all) interchromatin granules clusters and reaching, via Cajal bodies, the nucleolus. There, S6 is assembled with other proteins and rRNA into small ribosomal subunit (SSU), released in the nucleoplasm, and exported through the nuclear pores. We show for the first time the spatial association of nuclear myosin I (NMI) and actin with the SSU already at the nucleolar periphery to the nuclear pore. A blockade of NMI or actin induces an upstream accumulation of the S6 protein en route to the nucleolus, and a temperature lower than normal influences RNA export. Our data strongly suggest a functional relationship of SSU with NMI and actin. In our hypothesis, an active, myosin-driven movement of the small ribosomal subunit can be responsible for the export of approximately 10% of SSUs. This hypothesis is supported by ultrastructural, immunofluorescence, and biochemical analyses. The currently accepted model for the subunit release suggests a diffusive, temperature-independent mechanism. However, the advantage of the double mechanism would assure that the movement of a part of the subunits could be modulated, increased, or decreased according to the needs of the cell at a specific moment in the cell cycle.
- Published
- 2006
15. Glial Cell Reaction tocis-Dichlorodiammine Platinum Treatment in the Immature Rat Cerebellum
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Elda Scherini, Daniela Necchi, and Graziella Bernocchi
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Male ,Cerebellum ,Neurofilament ,Antineoplastic Agents ,Calbindin ,Purkinje Cells ,Developmental Neuroscience ,medicine ,Animals ,Rats, Wistar ,biology ,Glial fibrillary acidic protein ,GFAP stain ,Immunohistochemistry ,Rats ,Cell biology ,medicine.anatomical_structure ,Animals, Newborn ,nervous system ,Neurology ,biology.protein ,Phosphorylation ,Cisplatin ,Neuroglia ,Biomarkers ,Parvalbumin ,Immunostaining - Abstract
In this study we have investigated changes in glial cells of the cerebellum of cis-dichlorodiammine platinum (cisDDP)-treated rats. The expression of S-100 protein and glial fibrillary acidic protein (GFAP), taken as markers of glial cell function, was evaluated using immunocytochemical methods. In parallel, immunoreactivity for calbindin, parvalbumin, and phosphorylated 200-kDa neurofilament protein was observed in Purkinje cells as markers for neuronal integrity and activity. Results showed that, although no difference in the immunostaining of S-100 protein between control and treated animals could be observed, an increase in the frequency of GFAP immunoreactive cells was present in cisDDP-treated rats. In Purkinje cells, immunocytochemical expression of calbindin and parvalbumin was decreased after drug treatment. In addition, following immunoreaction for phosphorylated 200-kDa neurofilament protein, the somata of Purkinje cells, which were negative in control animals, were stained in treated rats. These findings suggest that cisDDP does not significantly interfere with pathways of glial cell activity and that the increased number of GFAP positive astrocytes may be due to an activation of glial cells consequent upon neuronal death.
- Published
- 1997
16. Zyxin is a novel target for β-amyloid peptide: characterization of its role in Alzheimer's pathogenesis
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Daniela Necchi, Maurizio Memo, Erica Buoso, Daniela Uberti, Antonella Pinto, Cristina Lanni, Stefano Govoni, Laura Buizza, and Marco Racchi
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Amyloid ,DNA damage ,Cell Survival ,Regulator ,Retinoic acid ,Protein Serine-Threonine Kinases ,Biochemistry ,Zyxin ,Cell Line ,methods ,antagonists /&/ inhibitors/metabolism ,Cellular and Molecular Neuroscience ,chemistry.chemical_compound ,Drug Delivery Systems ,Alzheimer Disease ,Cell Line, Tumor ,Amyloid precursor protein ,Humans ,Protein kinase A ,administration /&/ dosage ,Amyloid beta-Peptides ,antagonists /&/ inhibitors/physiology ,Tumor ,biology ,Protein Stability ,Signal transducing adaptor protein ,Protein-Serine-Threonine Kinases ,Peptide Fragments ,drug effects/physiology ,HEK293 Cells ,chemistry ,physiology ,biology.protein ,Cancer research ,etiology/metabolism/pathology, Amyloid beta-Peptides ,administration /&/ dosage, Carrier Proteins ,antagonists /&/ inhibitors/physiology, Cell Line ,Tumor, Cell Survival ,drug effects/physiology, Drug Delivery Systems ,methods, HEK293 Cells, Humans, Peptide Fragments ,administration /&/ dosage, Protein Stability, Protein-Serine-Threonine Kinases ,antagonists /&/ inhibitors/physiology, Signal Transduction ,physiology, Zyxin ,etiology/metabolism/pathology ,Carrier Proteins ,Signal Transduction - Abstract
Zyxin is an adaptor protein recently identified as a novel regulator of the homeodomain-interacting protein kinase 2 (HIPK2)-p53 signaling in response to DNA damage. We recently reported an altered conformational state of p53 in tissues from patients with Alzheimer ‘s disease (AD), because of a deregulation of HIPK2 activity, leading to an impaired and dysfunctional response to stressors. Here, we examined the molecular mechanisms underlying the deregulation of HIPK2 activity in two cellular models, HEK-293 cells and SH-SY5Y neuroblastoma cells differentiated with retinoic acid over-expressing the amyloid precursor protein, focusing on the evidence that zyxin expression is important to maintain HIPK2 protein stability. We demonstrated that both beta-amyloid (Aβ) 1-40 and 1-42 induce zyxin deregulation, thus affecting the transcriptional repressor activity of HIPK2 onto its target promoter, metallothionein 2A, which is in turn responsible for the induction of an altered conformational state of p53. We demonstrate for the first time that zyxin is a novel target of Aβ activities in AD. These results may help the studies on the pathogenesis of AD, through the fine dissection of events related to beta-amyloid activities.
- Published
- 2013
17. Abstracts Second Congress of the European Society for Clinical Neuropharmacology
- Author
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G. Collingridge, D. Bruns, A. Teufel, P. Barbier, H. G. Bloß, K. P. Offord, C. Stahl-Hennig, E. N. H. Jansen, T. Sobanski, Carlo Ori, M. Goetz, A. Muratorio, Ulderico Freo, E. Sale, Eldad Melamed, J. M. Maloteaux, Z. Bashir, B. Guschelbauer, D. Fitzgeral, R. Korinthenberg, H. Baas, Armin Heils, I. M. Macrae, T. Kutschka, M. C. Anderson, M. Beeg, G. A. Wiesbeck, P. Del Dotto, P. C. O'Brien, J. Knauber, M. E. Götz, Keith F. Tipton, M. Simanyi, G. Rossi, Max Holzer, I. Svobodová, L. Mancino, L. Calza, Th. Müller, J. P. W. F. Lakke, Mauro Dam, A. Raja, Eri Hashimoto, F. Crépel, U. Pulkowski, R. Ceravolo, D. R. Schroeder, M. Streifler, L. L. Iversen, E. Lossmann, K. Lieb, F. Heinen, T. A. Ibrahim, M. Rösier, F. Siebecker, R. Schinzel, P. Emson, A. H. Rajput, C. H. Lücking, F. Ferraguti, D. Feineis, L. Froelich, Wolf-Dieter Heiss, Ewout R. Brunt, Oliver Griesbeck, N. Pavese, A. Gerstner, J. Putzke, U. Nöth, Paolo Maria Rossini, R. God, G.B. Cassano, R. Nafc, D. Müller, A. Cunningham, Daniela Necchi, Patrick Carroll, C. Lucetti, F. Abel, O. M. Adegemo, E. Braak, B. Molzer, N. Fichter, A. Lugowska, M. L. Rao, S. Roßner, F. Coraddu, A. Gemma, O. Tucha, L. J. Bryan-Lluka, I. Avdiunina, H. Beckmann, P. Fruth, H. W. Clement, F. Müller, E. de la Morena, W. Zieglgänsberger, A. P. Jeanjean, C. H. Lammers, A. Seghers, A. Nuti, A. Steup, M. Schwarz, Michael Sendtner, T. W. van Weerden, M. Li, B. Janetzky, R. Erdmann, R. Winkel, B. Niedermeyer, V. ter Meulen, M. Butà, D. Peckys, Th. Arendt, P. A. Löschmann, S. Strauss, D. Offen, B. Gangus, D. M. Yilmazer, K. Velbinger, T. J. Feuerstein, Klaus V. Toyka, R. S. J. Frackowiak, F. Conquet, K. Gasiorowski, F. Fascetti, C. Grote, A. Barzilai, Thomas A. Sontag, C. G. Parsons, G. Dell’Agnello, Hans-Peter Hartung, Toshikazu Saito, R. Stein, W. D. Rausch, E. Donati, P. Vanhoorde, S. Hartmann, E. Orlov, D. Inglot, Francesca Vaglini, W. Paulus, A. Merico, W. H. Jost, H. H. Borchert, M. Bagli, N. S. Alekseeva, T. Heinemann, B. K. Siesjö, T. Hirning, I. Ziv, C. Wurthman, M. J. Lohse, E. Hermsteiner, J. Coos Verhoef, B. Landwehrmeyer, Félix F. Cruz-Sánchez, Hans Lassmann, R. Jackisch, E. W. Fünfgeld, M. Naumann, Gilberto Pizzolato, M. Bigl, Helmut Heinsen, J. E. Ahlskog, M. Sieklucka, Hiroki Ozawa, S. Hesse, J. Pruim, H. E. Junginger, Andreas Moser, J. Boning, F. Fumagalli, M. Berger, M. Lauk, E. Borroni, M. Gerlach, M. Heider, C. Schwarzer, K. Jellinger, W. H. Oertel, S. Danielcyk, V. Tuulik, J. Bauer, F. Block, Udo Rüb, F. Böcker, Hans Thoenen, L. Komelkova, G. Zürcher, A. Hochman, A. Mesec, G. Jungkunz, G. Charles, P. Vieregge, P. Kalus, Jürgen Fritze, I. Faé, K. Eschrich, H. Standhardt, M. Schüler, W. Kolasiewicz, A. Meister, E. N. H. Janzen, M Melzacka, A. A. Sharkawy, Borwin Bandelow, M. Renna, S. Hauck, Marco A. Riva, U. Lockemann, R. M. Nitsch, Heiko Braak, R. Medori, S. Federspiel, J. Berger, D. Senitz, J. Wissel, Georg W. Kreutzberg, U. McMonagle, H. Przuntek, T. Reum, C. Heim, K. V. Morgunov, R. Maggio, J. Leszek, Gavin P. Reynolds, Gerald Münch, M. Klessaschek, B. Zielke, R. Morgenstern, P. A. Fischer, Y. Agid, B. Volk, H. Schuttes, Konstanze Plaschke, J. Krieglstein, Th. Büttner, D. Blum-Degen, Eleni Koutsilieri, N. Wodarz, Reinhard Schliebs, P. König, Klaus W. Lange, T. Motzek-Noé, Robert Jech, J. Niemann, M. Abdel-moneim, V. V. Peresedov, Juergen Deckert, G. Storm, S. Kamolz, W. Breithaupt, B. Weber, Giovanni Corsini, J. C. Schwartz, M. Hüll, C. Bancher, M. Struck, M. Abdel-mohsen, A. Napolitano, D. Labunsky, M. Sohlbach, T. Winter, J. Sautter, H. J. Degen, Y. Glinka, R. Dörries, C. D. Earl, R. Riederer, G. Bringmann, W. Kuhn, J. A. Protzen, M. Winter, T. Klockgether, B. Fiebich, O. S. Brusov, H. Daniel, B. Bethke, T. R. Tolle, A. Weindl, Michael Bauer, N. Takahata, A. Baumer, Isabella Heuser, V. Gieselmann, Gian Franco Placidi, Giulio Perugi, H. Bönisch, A. Eckert, J. Michaelis, F. von Raison, V. Bigl, S. Harder, Graziella Bernocchi, J. Kuijpers, R. Brückner, U. Bonuccelli, M. José Barro, G. Laux, S. Grüter, W. Retz, M. L. Mimmack, A. Kupsch, Austin Smith, I. Zhirnova, A. M. Sardar, A. I. Svadovsky, Siegfried Hoyer, Peter Riederer, B. Haug, Thomas Arzberger, H. Bernheimer, M. Seemann, Karl-Heinz Sontag, D. Bengel, L. Demisch, W. Danielczyk, Bettina Holtmann, Ullrich Wüllner, E. Hermans, E. V. Khrapova, G. B. Landwehrmeyer, A. Tylki-Szymanska, R. Brinkmann, F. Remeš, B. Kanner, S. L. Timerbaeva, P. Piccini, Susanne Petri, W. Wesemann, G. Ulmar, F. Rausch, Leontino Battistin, U. Ziemann, H. B. Pollard, Gerhard Ransmayr, P. Mečíř, C. Mattern, U. Gärtner, S. Sopper, Moussa B.H. Youdim, Michael Deuschle, M. Pozza, H. Schubert, G. Goping, Rainer Spanagel, Lutz Frölich, L. HaveIec, Martina K. Brückner, W. Gsell, Werner Poewe, M. Da Prada, J. Hartmann, H. J. Stürenburg, B. Löschl, M. Norta, J. Dichgans, G. Stern, J. Mayr, Elda Scherini, D. Bleyl, A. Colzi, P. Rosario, C. D'Avino, J. X Xie, Klaus-Peter Lesch, M. Demuth, M. Ymamoto, A. Toso, K. Lehmann, F. Eblen, J. Thome, R. Burger, S. Šega, G. Farci, Evžen Růžička, F. W. H. M. Merkus, I. Strein, M. Rösler, T. Jaros, D. Barletta, W. W. Chan, U. Havemann-Reinecke, T. Kiauta, R. A. I. de Vos, S. Fähr, A. Körner, A. J. Willemsen, P. J. Neveu, A. V. Moshkin, A. Kleinschroth, L. A. Avdyuna, Johannes Kornhuber, Ryan J. Uitti, R. Häcker, Jan Roth, E. C. Laterre, H. Sternadl, Amos D. Korczyn, G. Künig, Werner E.G. Müller, W. Berger, G. Racagni, S. Salvetti, G. M. Emilien, Parsadanian As, K. Kunze, G. Sperk, D. Högemann, H. Maier, S. Behrens, D. K. Teherani, C Pardini, Karlheinz Reiners, T. S. Chen, C. J. Eggett, L. Popova, H. Reichmann, J. M. Rabey, H. Hartmann, Arvid Carlsson, B. Lawlor, F. Bürklin, O. Gleichauf, and S. Hemm
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0303 health sciences ,medicine.medical_specialty ,Public health ,03 medical and health sciences ,Psychiatry and Mental health ,0302 clinical medicine ,Neurology ,medicine ,Neurology (clinical) ,Psychiatry ,Psychology ,Neuroscience ,030217 neurology & neurosurgery ,Biological Psychiatry ,Neuropharmacology ,030304 developmental biology - Published
- 1995
18. Uneven distribution of NG2 cells in the rat cerebellar vermis and changes in aging
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Elda Scherini, Selene Lomoio, and Daniela Necchi
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Aging ,Cerebellum ,Histology ,cerebellum ,Immunocytochemistry ,Biophysics ,Biology ,Rats, Sprague-Dawley ,NG2 glia ,Cerebellar Cortex ,medicine ,Animals ,Distribution (pharmacology) ,Antigens ,lcsh:QH301-705.5 ,Brief Report ,Cell Biology ,Anatomy ,cerebellum, aging, NG2 glia ,Immunohistochemistry ,Cerebrocerebellum ,Rats ,Rat Cerebellum ,medicine.anatomical_structure ,nervous system ,lcsh:Biology (General) ,PROTEOGLYCAN 2 ,Cerebellar vermis ,Proteoglycans - Abstract
We describe by NG2 (neuron-glia chondroitin sulphate proteoglycan 2) immunocytochemistry an uneven distribution of NG2 glial cells in the rat cerebellum, being them more represented in the central lobules of the cerebellar vermis, belonging to the cerebrocerebellum. The cerebellar distribution of NG2 cells changes in aging rats, in which the area where the cells appear to be densely scattered throughout all cerebellar layers involves also more rostral and caudal lobules. In addition, in aging rats, in the most rostral and caudal lobules belonging to the spinocerebellum, punctate reaction product is present at the apical pole of Purkinje cells, i.e. in the area where the majority of synapses between olivary climbing fibers and Purkinje cells occur. Data suggest that the different distribution of NG2 cells is correlated to differences in physiology among cerebellar areas and reflects changes during aging.
- Published
- 2012
19. A single episode of neonatal seizures alters the cerebellum of immature rats
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Elda Scherini, Vladislav Mareš, Daniela Necchi, and Selene Lomoio
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medicine.medical_specialty ,Cerebellum ,Calbindins ,Immunocytochemistry ,Cell Count ,AMPA receptor ,Biology ,Tritium ,Calbindin ,Epilepsy ,Purkinje Cells ,S100 Calcium Binding Protein G ,Seizures ,Internal medicine ,medicine ,Animals ,Receptors, AMPA ,Axon ,Rats, Wistar ,Cell Proliferation ,Neuronal Plasticity ,Glutamate receptor ,medicine.disease ,Rats ,Disease Models, Animal ,Endocrinology ,medicine.anatomical_structure ,nervous system ,Neurology ,Animals, Newborn ,Excitatory Amino Acid Transporter 2 ,Cerebellar vermis ,Autoradiography ,Pentylenetetrazole ,Neurology (clinical) ,Thymidine - Abstract
Summary Purpose To test whether a single episode of early-life seizures may interfere with the development of the cerebellum. The cerebellum is particularly vulnerable in infants, since it is characterized by an important postnatal histogenesis that leads to the settling of adult circuitry. Methods Seizures were induced in 10-day-old Wistar rats with a single convulsive dose (80μg/g b.w., s.c.) of pentylentetrazole (PTZ). Immediately after rats were treated with 3 H-thymidine ( 3 HTdR, 2.5μCi/g b.w, s.c.). Rats were killed 4h later and paraffin sections of the cerebellar vermis were processed for 3 HTdR autoradiography and immunocytochemistry for 2/3 subunits of AMPA glutamate receptor (GluR2/3), glutamate transporter 1 (GLT1) and calbindin. Results Seizures reduced the proliferation rate of cells in the external germinal layer. Purkinje cells showed increased GluR2/3 immunoreactivity. However, some Purkinje cells were unstained or lost. Increased GLT1 immunoreactivity was present in glial cells surrounding Purkinje cells. Calbindin immunoreaction confirmed that some Purkinje cells were missed. The remaining Purkinje cells showed large spheroids along the course of their axon. Conclusions Data indicate that seizures lead to a loss and alteration of Purkinje cells in the cerebellum of immature rats. Since at 10 days of life Purkinje cells are no more proliferating, the loss of Purkinje cells should be permanent.
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- 2010
20. Morphological changes in the frog cerebellar cortex after unilateral section of the statoacustic nerve
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Graziella Bernocchi, Paolo Valli, Cristiana Soldani, Laura Botta, Daniela Necchi, and Elda Scherini
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Cytoplasm ,Cerebellum ,Histology ,medicine.medical_treatment ,Biophysics ,Cell Count ,Biology ,Functional Laterality ,Lesion ,Cerebellar Cortex ,Purkinje Cells ,Vestibular nuclei ,Neuroplasticity ,medicine ,Animals ,Tolonium Chloride ,Coloring Agents ,lcsh:QH301-705.5 ,Vestibular system ,Neuronal Plasticity ,Rana esculenta ,Neurectomy ,Cell Biology ,Anatomy ,Vestibular Nuclei ,Microscopy, Electron ,medicine.anatomical_structure ,lcsh:Biology (General) ,Cerebellar cortex ,medicine.symptom - Abstract
To investigate a possible role of the cerebellum in vestibular compensation that follows a lesion to the vestibular apparatus, the morphological changes of the cerebellar cortex of adult frogs following unilateral statoacustic nerve section was analyzed by means of electron microscopy starting from 3 days after the neurectomy for up to 6 months. On the ipsilateral side, massive abnormality was found in all layers at early postsurgical intervals. This involved both nerve fibers and cell bodies. Fibers often appeared condensed or vacuolated with poorly compacted myelin sheath. Cells had electronlucent and vacuolated cytoplasm to varying extent. Alterations became less conspicuous after 30 days and after 60 days altered nerve cells were no longer present. On the contralateral side, only a few Purkinje and granule cells were affected at early postsurgical stages. This may derive from the fact that, in the frog, some of the vestibular primary afferents reach contralateral cerebellar cortex. At 30 days, alterations had substantially progressed, and at 60 days they involved all the cortical layers. Fiber debris was present in the granular and molecular layers and numerous 317 Purkinje cells were electrondense and shrunken. This lateness in alteration may be a consequence of the prolonged silence of the vestibular nucleus contralateral to the lesion. At 4 and 6 months the tissue architecture was normal.
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- 2010
21. cis-Dichlorodiammineplatinum alters GABAergic structures in the immature rat cerebellum
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Graziella Bernocchi, Daniela Necchi, C. Vignola, and Elda Scherini
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medicine.medical_specialty ,Cerebellum ,Neurofilament ,Immunocytochemistry ,Purkinje cell ,Mitosis ,Biology ,Axon hillock ,Purkinje Cells ,Pericellular basket ,Neurofilament Proteins ,Internal medicine ,medicine ,Animals ,Rats, Wistar ,Axon ,gamma-Aminobutyric Acid ,Glutamate Decarboxylase ,General Neuroscience ,Body Weight ,Organ Size ,Rats ,Molecular Weight ,medicine.anatomical_structure ,Endocrinology ,GABAergic ,Cisplatin - Abstract
It has been reported that injection of the antitumoral drug cis -dichlorodiammineplatinum at 10 days of life affects cerebellar development in rats. After a single dose of 5 μg/g of body weight, the formation of granule cells is decreased and the maturation of postmitotic neurons is slowed down. A substantial time after treatment, reduced cell packing density of the internal granule layer and atrophy of the molecular layer can be observed. In addition, there is degeneration of some Purkinje cells and Golgi neurons. In spite of all these alterations, the regular architecture of the cerebellar folia is retained in many places. In the present study, we used immunocytochemistry with an immune serum raised against glutamic acid decar☐ylase to further characterize the cis -dichlorodiammineplatinum-induced alterations of GABAergic neurons. The aim was to examine cerebellar development and to test for factors controlling the settling of GABAergic circuits. At all post-treatment intervals, most of the Purkinje and Golgi neurons and molecular layer interneurons showed stronger anti-glutamic acid decar☐ylase immunoreactivity than in controls; this may have been due to altered fixation because of cis -dichlorodiammineplatinum-induced damages to the blood vessels; but could also reflect cellular retention of the enzyme, maybe due to cis -dichlorodiammineplatinum-induced damage of the microtubular apparatus. After seven days, large roundish immunoreactive varicosities were present in the molecular layer adjacent to the Purkinje cell dendritic poles. These varicosities, which were not observed in control animals, may be terminals of Purkinje cell axon recurrent collaterals contributing to the supraganglionic plexus, whose abnormal development would compensate for the reduced inhibitory inputs from inhibitory interneurons and/or Purkinje cells, which degenerated at early post-treatment intervals. At later post-treatment intervals (15 and 21 days), there were also alterations in the pericellular basket at the Purkinje cell axon hillock, which was poorly developed in or absent from the majority of cells. The finding was confirmed by morphological observation of basket cells in Golgi-Cox preparation and immunocytochemistry with an antibody raised against 200,000 mol. wt phosphorylated neurofilaments. It is concluded that early changes in anti-glutamic acid decar☐ylase immunoreactivity of neurons may be due to a direct interference of the drug with the cellular metabolic pathways. The late anomalies in the anti-glutamic acid decar☐ylase immunoreactivity appear to be secondary to changes in the tissue cytoarchitecture rather than being primary cis -dichlorodiammineplatinum-induced lesions of the cells.
- Published
- 1992
22. Degradation of p21CDKN1A after DNA damage is independent of type of lesion, and is not required for DNA repair
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Ornella Cazzalini, Paola Perucca, Monica Savio, Lucia Anna Stivala, Ennio Prosperi, Tiziana Nardo, Tania Coppa, and Daniela Necchi
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Cyclin-Dependent Kinase Inhibitor p21 ,Time Factors ,DNA Repair ,DNA damage ,DNA repair ,Cell Survival ,Ultraviolet Rays ,Blotting, Western ,Fluorescent Antibody Technique ,Biology ,Transfection ,Biochemistry ,DNA polymerase delta ,Cell Line ,Proliferating Cell Nuclear Antigen ,Humans ,Molecular Biology ,Replication protein A ,Cells, Cultured ,Cell Biology ,DNA Repair Pathway ,DNA repair protein XRCC4 ,Fibroblasts ,Molecular biology ,Cross-Linking Reagents ,Mutation ,DNA mismatch repair ,Cisplatin ,Nucleotide excision repair ,DNA Damage ,HeLa Cells ,Protein Binding ,Signal Transduction - Abstract
The inhibitor of cyclin-dependent kinases p21CDKN1A plays a fundamental role in several pathways involved in the DNA damage response, like checkpoint-mediated cell cycle arrest, transcription, apoptosis, and DNA repair. Although p21 protein level is regulated by proteasomal degradation, the relationship of this process with DNA repair pathways is not yet clear. In addition, the role of protein/protein interaction in regulating turnover of p21 protein, is controversial. Here, we show that in human fibroblasts treated with agents inducing lesions repaired through nucleotide excision repair (NER), or base excision repair (BER), p21 degradation was triggered more by the extent, than by the type of DNA damage, or consequent DNA repair pathway. In fact, lowering the amount of DNA damage resulted in an increased stability of p21 protein. Overexpression of p21 was found to obscure degradation, both for p21wt and a p21 mutant unable to bind PCNA (p21PCNA-). However, when expressed to lower levels, turnover of p21 protein after DNA damage was greatly influenced by interaction with PCNA, since p21PCNA- was more efficiently degraded than wild-type protein. Interestingly, a p21 mutant protein unable to localize in the nucleus because of mutations in the NLS region, was not degraded after DNA damage, thus indicating that nuclear localization is necessary for p21 turnover. Removal of p21 was not required for NER activity, since inhibition of p21 degradation by caffeine did not affect the UV-induced recruitment of repair proteins, such as PCNA and DNA polymerase delta, nor significantly influence DNA repair synthesis, as determined by autoradiography. These results indicate that degradation of p21 is not dependent on a particular DNA repair pathway, and is not required for efficient DNA repair.
- Published
- 2008
23. Axonal abnormalities in cerebellar Purkinje cells of the Ts65Dn mouse
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Daniela Necchi, Elda Scherini, and Selene Lomoio
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Cerebellum ,Pathology ,medicine.medical_specialty ,Immunocytochemistry ,Central nervous system ,Purkinje cell ,Granular layer ,Biology ,Calbindin ,Cerebellar Cortex ,Mice ,Purkinje Cells ,Genetic model ,medicine ,Image Processing, Computer-Assisted ,Animals ,Axon ,Molecular Biology ,General Neuroscience ,Immunohistochemistry ,Axons ,Disease Models, Animal ,medicine.anatomical_structure ,nervous system ,Female ,Neurology (clinical) ,Down Syndrome ,Neuroscience ,Developmental Biology - Abstract
Ts65Dn mice are a genetic model for Down syndrome. Among others, these mice have cerebellar pathology features which parallel those seen in Down syndrome patients. Both individuals with Down syndrome and Ts65Dn mice have reduced cerebellar volume and numbers of granule and Purkinje cells. In this report, we describe morphological abnormalities of axons of Purkinje cells in the cerebellum of Ts65Dn mice, by using anti-calbindin immunocytochemistry. A consistent number of Purkinje cells shows axons bearing giant varicosities along their transit through the granular layer. The cerebellar arbor vitae made by fasciculated Purkinje cell axons has a patchy appearance, some tracks being devoid of calbindin staining. The infraganglionic plexus, formed by recurrent collaterals of Purkinje cell axons, has enormously increased density, which is evidence for a compensatory reaction to degeneration of distal segments of axons. These alterations are accompanied by strong glial reaction as evidenced by GFAP immunocytochemistry. Moreover, the alterations are more consistent in the anterior lobules of the vermis and intermediate cortex. The axonal pathology of Purkinje cells may explain the impairment in cerebellar functions observed in Ts65Dn mice at the adulthood.
- Published
- 2008
24. Both early and delayed treatment with melanocortin 4 receptor-stimulating melanocortins produces neuroprotection in cerebral ischemia
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Sheila Leone, Davide Zaffe, Daniela Necchi, Alfio Bertolini, Chiara Mioni, Letteria Minutoli, Helgi B. Schiöth, Francesco Squadrito, Daniela Giuliani, Alessandra Bitto, Domenica Altavilla, Salvatore Guarini, Annibale R. Botticelli, Carla Bazzani, Roberto Pizzala, Herbert Marini, Maria Michela Cainazzo, and Monica Savio
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Central Nervous System ,Male ,Cytoplasm ,Time Factors ,Apoptosis ,ALPHA-MSH ,Hippocampus ,ACTIVATED PROTEIN-KINASES ,PITUITARY-ADRENAL-AXIS ,TUMOR-NECROSIS-FACTOR ,TRANSIENT FOREBRAIN ISCHEMIA ,HEMORRHAGE-SHOCKED RATS ,BCL-2 FAMILY PROTEINS ,BLOOD-BRAIN-BARRIER ,FACTOR-KAPPA-B ,GERBIL HIPPOCAMPUS ,Brain Ischemia ,Brain ischemia ,Endocrinology ,Ischemia ,Hypoxia ,bcl-2-Associated X Protein ,Neurons ,Caspase 3 ,Brain ,Melanocortin 4 receptor ,Stroke ,Neuroprotective Agents ,Treatment Outcome ,Caspases ,Cytokines ,Receptor, Melanocortin, Type 4 ,Melanocortin ,Agonist ,medicine.medical_specialty ,medicine.drug_class ,MAP Kinase Signaling System ,Blotting, Western ,bcl-X Protein ,Neuroprotection ,Melanocortin receptor ,Memory ,Internal medicine ,medicine ,Animals ,Learning ,Maze Learning ,Melanocortins ,Inflammation ,Models, Statistical ,business.industry ,medicine.disease ,Enzyme Activation ,Disease Models, Animal ,alpha-MSH ,business ,Gerbillinae ,DNA Damage - Abstract
Ischemic stroke is one of the main causes of death and disability. We investigated whether melanocortin peptides, which have protective effects in severe hypoxic conditions, also produce neuroprotection in a gerbil model of ischemic stroke. A 10-min period of global cerebral ischemia, induced by occluding both common carotid arteries, caused impairment in spatial learning and memory that was associated with activation of inflammatory and apoptotic pathways, including severe DNA damage and delayed neuronal death, in the hippocampus. Treatment with nanomolar doses of the melanocortin analog [Nle4, D-Phe7] alpha-MSH [which activates the melanocortin receptor subtypes (MC) mainly expressed in central nervous system, namely MC3 and MC4] modulated the inflammatory and apoptotic cascades and reduced hippocampus injuries even when delayed up to 9 h after ischemia, with consequent dose-dependent improvement in subsequent functional recovery. The selective MC3 receptor agonist gamma2-MSH had no protective effects. Pharmacological blockade of MC4 receptors prevented the neuroprotective effects of [Nle4, D-Phe7] alpha-MSH and worsened some ischemia outcomes. Together, our findings suggest that MC4 receptor-stimulating melanocortins might provide potential to develop a class of drugs with a broad treatment window for a novel approach to neuroprotection in ischemic stroke.
- Published
- 2006
25. MPTP-induced increase in c-Fos- and c-Jun-like immunoreactivity in the monkey cerebellum
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Daniela, Necchi, C, Soldani, F, Ronchetti, G, Bernocchi, and E, Scherini
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Male ,Macaca fascicularis ,Purkinje Cells ,1-Methyl-4-phenyl-1,2,3,6-tetrahydropyridine ,Proto-Oncogene Proteins c-jun ,Cerebellum ,Dopamine Agents ,Animals ,Immunohistochemistry ,Proto-Oncogene Proteins c-fos ,Up-Regulation - Abstract
The transcription factors c-Fos and c-Jun have been described to be overexpressed following many pathological stimuli, but whether they are required for neurodegeneration or neuroprotection is still open. In the present report, we analyzed the role of c-Fos and c-Jun proteins in Purkinje cell degeneration caused by the neurotoxin MPTP (1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine) in the monkey cerebellum, and determined the neuroprotective effect of the antioxidant drug a-dihydroergocryptine (DHEC), whose prior and simultaneous administration reduced the MPTP-induced neuronal loss in the substantia nigra. Immunocytochemistry for c-Fos- and c-Jun-like proteins showed persistent increased staining in Purkinje cells of MPTP-treated monkeys. The staining was greatly reduced in animals receiving DHEC. Similar results were observed in white matter glial cells after immunoreaction for c-Fos. The results suggest that, at least as far as the cerebellum is concerned, the increase in c-Fos and c-Jun expression correlate with cell damage, rather than with preservation.
- Published
- 2005
26. Regional alterations of the NO/NOS system in the aging brain: a biochemical, histochemical and immunochemical study in the rat
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Antonio Contestabile, Barbara Monti, Marco Virgili, Elda Scherini, and Daniela Necchi
- Subjects
Olfactory system ,Male ,Pathology ,medicine.medical_specialty ,Aging ,Central nervous system ,Blotting, Western ,Hippocampus ,Cell Count ,Striatum ,Nitric Oxide Synthase Type I ,Biology ,Nitric Oxide ,Biochemistry ,Nitric oxide ,chemistry.chemical_compound ,Internal medicine ,medicine ,Aging brain ,Animals ,Rats, Wistar ,Molecular Biology ,Neurons ,Neocortex ,Histocytochemistry ,General Neuroscience ,Immunochemistry ,NADPH Dehydrogenase ,Brain ,Rats ,Nitric oxide synthase ,medicine.anatomical_structure ,Endocrinology ,nervous system ,chemistry ,biology.protein ,Neurology (clinical) ,Nitric Oxide Synthase ,Developmental Biology - Abstract
We have used several approaches (immunohistochemistry and enzyme histochemistry, Western blotting, biochemical assay of Ca(2+)-dependent catalytic activity) in order to detect differences in neuronal nitric oxide synthase (nNOS) expression and activity in various brain regions of young-adult (4-month-old) and aged (28-month-old) rats. In most of the brain regions examined (striatum, neocortex, olfactory cortex and hippocampus) some significant decrease in the density per unit area of nNOS neurons, detected either through immunohistochemistry or enzyme histochemistry, was observed in aged rats. However, only in the striatum and olfactory cortex this was accompanied by a significant decrease of the catalytic activity of the constitutive, Ca(2+)-dependent NOS form. In these two regions, the relative level of expression of nNOS protein was also significantly decreased, as assessed by Western blotting of proteic extracts from young-adult and aged rats. Other observed differences were a paler stain of neurons in some brain areas of the aged rats and differences of cellular compartmentalization of the protein in the same rats, as assessed through confocal microscopy. The present observations demonstrate that the expression and activity of nNOS show regionally-specific alterations in the brain of aged healthy rats, with a trend towards decrease, rather than toward increase as suggested by some previous reports. Therefore, hypotheses implicating nitric oxide increase in brain aging should be reconsidered on the basis of a clear-cut distinction between the physiological and the pathological aspects of the aging process.
- Published
- 2002
27. Abnormal vestibular control of gaze and posture in a strain of a waltzing rat
- Author
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Daniela Necchi, G Rabbath, J.-P. Gasc, P.P. Vidal, P. Josset, and C. de Waele
- Subjects
Male ,genetic structures ,Rotation ,Posture ,Fixation, Ocular ,S100 Calcium Binding Protein G ,Neurofilament Proteins ,otorhinolaryngologic diseases ,medicine ,Animals ,Vestibular system ,Movement Disorders ,Semicircular canal ,General Neuroscience ,Motor control ,Rats, Inbred Strains ,Anatomy ,Reflex, Vestibulo-Ocular ,Vestibular nerve ,Immunohistochemistry ,Semicircular Canals ,Rats ,Microscopy, Electron ,medicine.anatomical_structure ,Vestibule ,Calbindin 2 ,Fixation (visual) ,Reflex ,Female ,sense organs ,Vestibule, Labyrinth ,Vestibulo–ocular reflex ,Psychology ,Neuroscience - Abstract
The waltzing behavior is usually attributed to vestibular dysfunction. However, the vestibular control of gaze and posture has not yet been measured quantitatively in any waltzing mutant. Therefore, this study was aimed at investigating the relationship between inner-ear morphology, the circling behavior, and the vestibular control of gaze and posture in a new strain of waltzing rats. Light- and electron-microscopy studies of these mutants did not reveal any structural abnormalities of the vestibular neural epithelia. In addition, the expression of Calretinin and 200-kD phosphorylated and non-phosphorylated neurofilaments was also found to be normal in the vestibular neural epithelia and ganglion cells. In contrast, the mutants showed severe dysfunctions of the vestibular control of gaze and posture. The skeletal geometry of the alert unrestrained animals was studied using cineradiography. At rest, waltzing rats held their heads tilted down: the horizontal semicircular-canal's plane was near the earth-horizontal's plane, instead of being tilted up as in Long Evans control rats. In addition, their cervical column was pitched more forward (33.6 degrees) than in the control group (6.9 degrees). The circling behavior was observed frequently, and the rats had episodes of circling in both directions. The episodes of circling amounted to an average of 17 turns, and the average angular velocity of the circling was 645 degrees/s. Unilateral labyrinthectomy induced the same postural and oculomotor syndromes in the waltzing and control groups. This indicates that the mutant vestibular nerve had a significant resting discharge before the lesion. Eye movements were recorded using acutely implanted search coils. Although waltzing rats were able to perform normal spontaneous eye movements, they showed a complete deficit of the horizontal vestibulo-ocular reflex (HVOR) and an impairment of the maculo-ocular reflex (MOR) during constant velocity, off-vertical axis rotation (OVAR). These results show, for the first time, that deficient transduction and/or processing of the horizontal-canal- and macular-related information can be causally related to the circling behavior and abnormal posture, respectively.
- Published
- 2001
28. Development of the anatomical alteration of the cerebellar fissura prima
- Author
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Graziella Bernocchi, Elda Scherini, Cristiana Soldani, and Daniela Necchi
- Subjects
Cerebellum ,Pia mater ,Staining and Labeling ,Granule (cell biology) ,External Granular Layer ,Anatomy ,Biology ,Histogenesis ,Granule cell ,Agricultural and Biological Sciences (miscellaneous) ,Immunohistochemistry ,Rats ,Brush Cell ,Purkinje Cells ,medicine.anatomical_structure ,S100 Calcium Binding Protein G ,Calbindin 2 ,medicine ,Animals ,Calretinin ,Rats, Wistar - Abstract
The development of the naturally occurring malformation of the cerebellar fissura prima was monitored in rats starting from 4 days of life to the adulthood. The first sign of the malformation was evident at 10 days of life and consisted of an interruption of the pia mater and the fusion of the external granular layers on the two sides of the fissura. Later, nests of apparently mature granule cells could be seen to be encircled by cells of the external granular layer and to be connected to the granule cell layer by thin bridges of cells. Calretinin immunoreactive fibers followed the bridges of cells to reach the ectopic masses of cells. Towards the end of histogenesis and in adult animals, brush cells and Golgi cells were present in the ectopic masses of granule cells. The latter appeared to contribute to the formation of normal glomeruli, as in the orthotopic granule cell layer. In addition, bundles of parallel fibers crossed the boundary between the molecular layers on the two side of the fissure, thus suggesting that parallel fibers can contact Purkinje cells of the opposite folium.
- Published
- 2000
29. Perineuronal glial system in the cerebral ganglion of active and hibernating Helix aspersa
- Author
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Daniela Necchi, Carla Fenoglio, Cristiana Soldani, Elda Scherini, and Graziella Bernocchi
- Subjects
Functional role ,biology ,Helix (gastropod) ,ATPase ,Period (gene) ,Land snail ,Cell Biology ,General Medicine ,Anatomy ,biology.organism_classification ,Cell biology ,nervous system ,Extracellular ,biology.protein ,Cerebral ganglion ,Alkaline phosphatase ,Developmental Biology - Abstract
The electron microscopical changes in the glial lacunar network that surrounds the large neurons of meso- and metacerebrum of land snail cerebral ganglia were considered, in order to get an insight into the functional role of this peculiar structure in invertebrates. Compared with snails during the active period, in the hibernating animals the extension of glial processes was reduced and the glial processes appeared more regular and stacked around neurons. Moreover, they did not form deep, long interdigitations with neuronal infoldings as during the active period. In particular, data on the ultracytochemical detection of alkaline phosphatase and Ca 2+ /Mg 2+ -ATPase enzyme activities, point to a correlation between the extension of the glial system and its function in the regulation of the extracellular environment. In fact, in hibernating snails, lower reactivity was found on the glial membranes, including those of the trophospongium.
- Published
- 1997
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