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2 results on '"MESH : P-Glycoprotein"'

1. Expression of drug transporters at the blood–brain barrier using an optimized isolated rat brain microvessel strategy

Author

Salah Yousif, Françoise Roux, Xavier Declèves, Cynthia Marie-Claire, Jean-Michel Scherrmann, Neuropsychopharmacologie des addictions. Vulnérabilité et variabilité expérimentale et clinique (NAVVEC - UM 81 (UMR 8206/ U705)), Centre National de la Recherche Scientifique (CNRS)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Université Paris Descartes - Paris 5 (UPD5)-Institut des sciences du Médicament -Toxicologie - Chimie - Environnement (IFR71), Institut National de la Santé et de la Recherche Médicale (INSERM)-Ecole Nationale Supérieure de Chimie de Paris - Chimie ParisTech-PSL (ENSCP), Université Paris sciences et lettres (PSL)-Université Paris sciences et lettres (PSL)-Centre National de la Recherche Scientifique (CNRS)-Institut de Recherche pour le Développement (IRD)-Université Paris Descartes - Paris 5 (UPD5)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Ecole Nationale Supérieure de Chimie de Paris - Chimie ParisTech-PSL (ENSCP), Université Paris sciences et lettres (PSL)-Université Paris sciences et lettres (PSL)-Centre National de la Recherche Scientifique (CNRS)-Institut de Recherche pour le Développement (IRD)-Université Paris Descartes - Paris 5 (UPD5)-Université Paris Diderot - Paris 7 (UPD7), Université Paris Diderot - Paris 7 (UPD7)-Institut des sciences du Médicament -Toxicologie - Chimie - Environnement (IFR71), Institut de Recherche pour le Développement (IRD)-Ecole Nationale Supérieure de Chimie de Paris - Chimie ParisTech-PSL (ENSCP), Université Paris sciences et lettres (PSL)-Université Paris sciences et lettres (PSL)-Université Paris Descartes - Paris 5 (UPD5)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Centre National de la Recherche Scientifique (CNRS)-Institut de Recherche pour le Développement (IRD)-Ecole Nationale Supérieure de Chimie de Paris - Chimie ParisTech-PSL (ENSCP), Université Paris sciences et lettres (PSL)-Université Paris sciences et lettres (PSL)-Université Paris Descartes - Paris 5 (UPD5)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Centre National de la Recherche Scientifique (CNRS)-Université Paris Descartes - Paris 5 (UPD5)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Centre National de la Recherche Scientifique (CNRS), Marie-Claire, Cynthia, Neuropsychopharmacologie des addictions. Vulnérabilité et variabilité expérimentale et clinique ( NAVVEC (UM 81) ), Université Paris Diderot - Paris 7 ( UPD7 ) -Institut des sciences du Médicament -Toxicologie - Chimie - Environnement ( IFR71 ), and Institut National de la Santé et de la Recherche Médicale ( INSERM ) -Ecole Nationale Supérieure de Chimie de Paris- Chimie ParisTech-PSL ( ENSCP ) -Centre National de la Recherche Scientifique ( CNRS ) -Institut de Recherche pour le Développement ( IRD ) -Université Paris Descartes - Paris 5 ( UPD5 ) -Institut National de la Santé et de la Recherche Médicale ( INSERM ) -Ecole Nationale Supérieure de Chimie de Paris- Chimie ParisTech-PSL ( ENSCP ) -Centre National de la Recherche Scientifique ( CNRS ) -Institut de Recherche pour le Développement ( IRD ) -Université Paris Descartes - Paris 5 ( UPD5 ) -Université Paris Descartes - Paris 5 ( UPD5 ) -Institut National de la Santé et de la Recherche Médicale ( INSERM ) -Centre National de la Recherche Scientifique ( CNRS )

Subjects
Male, MESH : RNA, Messenger, MESH : Multidrug Resistance-Associated Proteins, MESH: Rats, Sprague-Dawley, MESH : Microcirculation, MESH : P-Glycoprotein, MESH: Membrane Transport Proteins, MESH : Blood-Brain Barrier, Rats, Sprague-Dawley, MESH: Blood-Brain Barrier, 0302 clinical medicine, MESH : Membrane Transport Proteins, MESH: Microcirculation, Gene expression, MESH: Glial Fibrillary Acidic Protein, MESH: Animals, MESH: Endothelial Cells, Microvessel, MESH: Cerebral Arteries, MESH : Organ Culture Techniques, 0303 health sciences, MESH : Gene Expression Regulation, biology, MESH : Rats, General Neuroscience, Isolated brain, MESH : Glial Fibrillary Acidic Protein, MESH: Gene Expression Regulation, Multidrug Resistance-Associated Protein 2, Blot, medicine.anatomical_structure, [ SDV.BBM.GTP ] Life Sciences [q-bio]/Biochemistry, Molecular Biology/Genomics [q-bio.GN], Blood-Brain Barrier, cardiovascular system, [SDV.BBM.GTP] Life Sciences [q-bio]/Biochemistry, Molecular Biology/Genomics [q-bio.GN], MESH : Carrier Proteins, Multidrug Resistance-Associated Proteins, MESH: Multidrug Resistance-Associated Proteins, Astrocyte, MESH: P-Glycoprotein, MESH: Rats, MESH : Male, MESH: Carrier Proteins, Blood–brain barrier, 03 medical and health sciences, Organ Culture Techniques, [SDV.BBM.GTP]Life Sciences [q-bio]/Biochemistry, Molecular Biology/Genomics [q-bio.GN], Glial Fibrillary Acidic Protein, medicine, Animals, ATP Binding Cassette Transporter, Subfamily B, Member 1, RNA, Messenger, Molecular Biology, MESH: RNA, Messenger, 030304 developmental biology, MESH : Endothelial Cells, MESH : Cerebral Arteries, Microcirculation, MESH: Biological Markers, Endothelial Cells, Membrane Transport Proteins, Cerebral Arteries, Molecular biology, MESH : Rats, Sprague-Dawley, MESH: Organ Culture Techniques, MESH: Male, Rats, MESH : Biological Markers, Gene Expression Regulation, nervous system, Synaptophysin, biology.protein, MESH : Animals, Neurology (clinical), Neuron, Carrier Proteins, Biomarkers, 030217 neurology & neurosurgery, Developmental Biology
Abstract

International audience; Quantitative RT-PCR (qRT-PCR) and Western blotting studies on transporters at the blood-brain barrier (BBB) of isolated brain microvessels have produced conflicting data on their cellular distribution. A major problem is identifying cells expressing the genes of interest, since isolated brain microvessels are composed of several cell types and may be contaminated with mRNA or proteins from astrocytes and neurons. We isolated rat brain microvessels and examined microscopically samples at each step of isolation to evaluate microvessel purity. The expression of specific markers of endothelial cells (Glut-1, Flk-1), pericytes (Ng2), neurons (synaptophysin, Syn) and astrocytes (Gfap) was measured by qRT-PCR in order to select the protocol giving the least astrocyte and neuron mRNAs and the most endothelial mRNAs. We also evaluated the gene expression of drug transporters (Mdr1a, Mdr1b, Mrp1-5, Bcrp and Oatp-2) at each step to optimize their location in cells at the BBB. The Mdr1a, Mrp4, Bcrp and Oatp-2 gene profiles were similar to those of endothelium markers. The profiles of Mrp2 and Mrp3 closely resembled that of Ng2. Mrp5 and Mrp1 expression was not increased in the microvessel-enriched fraction, suggesting that they are ubiquitously expressed throughout the cortex parenchyma. We also evaluated by Western blotting the expression of P-gp, Mrp2, Gfap and Syn in the cortex and in the purest obtained microvessel fraction. Our results showed that P-gp expression strongly increased in microvessels whereas Mrp2 was not detected in any of the fraction. Surprisingly, Gfap expression increased in isolated microvessels whereas Syn was not detected. Our results showed that the strategy consisting of identifying gene expression at different steps of the protocol is useful to identify cells containing mRNA at the BBB and give overall similar results with protein expression.

Published
2007
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2. Expression and differential localization of xenobiotic transporters in the rat olfactory neuro-epithelium

Author

Yves Artur, Christine Belloir, Nicolas Thiebaud, Anne-Marie Le Bon, Jean-Marie Heydel, Franck Ménétrier, Anne-Laure Minn, Fabrice Neiers, Centre des Sciences du Goût et de l'Alimentation [Dijon] (CSGA), Institut National de la Recherche Agronomique (INRA)-Université de Bourgogne (UB)-AgroSup Dijon - Institut National Supérieur des Sciences Agronomiques, de l'Alimentation et de l'Environnement-Centre National de la Recherche Scientifique (CNRS), Centre National de la Recherche Scientifique (CNRS)-AgroSup Dijon - Institut National Supérieur des Sciences Agronomiques, de l'Alimentation et de l'Environnement-Institut National de la Recherche Agronomique (INRA)-Université de Bourgogne (UB), Agence Nationale de la Recherche (ANR-05-PNRA-1.E7), Aromalim Burgundy Regional Council (IFR92), French Ministry of Research, Centre des Sciences du Goût et de l'Alimentation [Dijon] ( CSGA ), and Institut National de la Recherche Agronomique ( INRA ) -Université de Bourgogne ( UB ) -AgroSup Dijon - Institut National Supérieur des Sciences Agronomiques, de l'Alimentation et de l'Environnement-Centre National de la Recherche Scientifique ( CNRS )

Subjects
Male, [ SDV.AEN ] Life Sciences [q-bio]/Food and Nutrition, MESH : Multidrug Resistance-Associated Proteins, p glycoprotein, ATP-binding cassette transporter, MESH : Hepatocytes, Receptors, Odorant, MESH : P-Glycoprotein, MESH: Hepatocytes, MESH : Lymphatic Vessels, 0302 clinical medicine, MESH : Protein Transport, ugt2a1, MESH: Smell, MESH: Receptors, Odorant, MESH: Animals, Receptor, xenobiotic metabolizing, mucosa, 0303 health sciences, MESH : Gene Expression Regulation, MESH : Rats, General Neuroscience, MESH : Odors, MESH: Gene Expression Regulation, Smell, Protein Transport, medicine.anatomical_structure, Biochemistry, Liver, transporter, barrier, Efflux, Multidrug Resistance-Associated Proteins, MESH: Multidrug Resistance-Associated Proteins, MESH: Xenobiotics, MESH: Protein Transport, MESH: P-Glycoprotein, MESH: Rats, MESH: Lymphatic Vessels, MESH : Male, odorant clearance, brain, MESH : Xenobiotics, xenobiotic, Biology, system, MESH : Rats, Wistar, Olfactory Receptor Neurons, Xenobiotics, 03 medical and health sciences, bulb, Olfactory Mucosa, multidrug resistance, MESH : Receptors, Odorant, medicine, Animals, ATP Binding Cassette Transporter, Subfamily B, Member 1, Rats, Wistar, detoxification, MESH: Olfactory Mucosa, 030304 developmental biology, Lymphatic Vessels, MESH : Olfactory Mucosa, MESH: Odors, MESH : Liver, Transporter, MESH: Rats, Wistar, MESH: Olfactory Receptor Neurons, Epithelium, MESH: Male, Olfactory bulb, Rats, enzyme, Gene Expression Regulation, Odorants, Hepatocytes, MESH : Smell, MESH : Olfactory Receptor Neurons, MESH : Animals, olfactory epithelium, Olfactory epithelium, perireceptor event, [SDV.AEN]Life Sciences [q-bio]/Food and Nutrition, 030217 neurology & neurosurgery, Drug metabolism, MESH: Liver
Abstract

International audience; Transporters, such as multidrug resistance P-glycoproteins (MDR), multidrug resistance-related proteins (MRP) and organic anion transporters (OATs), are involved in xenobiotic metabolism, particularly the cellular uptake or efflux of xenobiotics (and endobiotics) or their metabolites. The olfactory epithelium is exposed to both inhaled xenobiotics and those coming from systemic circulation. This tissue has been described as a pathway for xenobiotics to the brain via olfactory perineural space. Thereby, olfactory transporters and xenobiotic metabolizing enzymes, dedicated to the inactivation and the elimination of xenobiotics, have been involved in the toxicological protection of the brain, the olfactory epithelium itself and the whole body. These proteins could also have a role in the preservation of the olfactory sensitivity by inactivation and clearance of the excess of odorant molecules from the perireceptor space. The goal of the present study was to increase our understanding of the expression and the localization of transporters in this tissue. For most of the studied transporters, we observed an opposite mRNA expression pattern (RT-PCR) in the olfactory epithelium compared to the liver, which is considered to be the main metabolic organ. Olfactory epithelium mainly expressed efflux transporters (MRP, MDR). However, a similar pattern was observed between the olfactory epithelium and the olfactory bulb. We also demonstrate distinct cellular immunolocalization of the transporters in the olfactory epithelium. As previously reported, Mrp1 was mainly found in the supranuclear portions of supporting cells. In addition, Mrp3 and Mrp5 proteins, which were detected for the first time in olfactory epithelium, were localized to the olfactory neuron layer, while Mdr1 was localized to the capillary endothelium of lymphatic vessels in the subepithelial region. The pattern of expression and the distinct localization of the olfactory transporters showed in this work may highlight on their specific function in the whole olfactory epithelium.

Published
2011
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