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27 results on '"MESH : Calcium"'

1. Genetic and functional characterisation of the lactococcal P335 phage-host interactions

Author

Laurens Hanemaaijer, Jennifer Mahony, Joana Oliveira, Marco Ventura, Douwe van Sinderen, Horst Neve, Gabriele Andrea Lugli, Barry Collins, Christian Cambillau, Thijs R. H. M. Kouwen, School of Microbiology, University College Cork (UCC), Università degli studi di Parma [Parme, Italie], Dept Microbiol & Biotechnol, Max Rubner Inst, Architecture et fonction des macromolécules biologiques (AFMB), Institut National de la Recherche Agronomique (INRA)-Aix Marseille Université (AMU)-Centre National de la Recherche Scientifique (CNRS), Department of Microbiology, University College Cork, Università degli studi di Parma = University of Parma (UNIPR), University of Parma = Università degli studi di Parma [Parme, Italie], Centre National de la Recherche Scientifique (CNRS)-Aix Marseille Université (AMU)-Institut National de la Recherche Agronomique (INRA), University College Cork ( UCC ), Università degli studi di Parma, Architecture et fonction des macromolécules biologiques ( AFMB ), and Centre National de la Recherche Scientifique ( CNRS ) -Aix Marseille Université ( AMU ) -Institut National de la Recherche Agronomique ( INRA )

Subjects
0301 basic medicine, viruses, [ SDV.MP.BAC ] Life Sciences [q-bio]/Microbiology and Parasitology/Bacteriology, Proteomics, Siphoviridae, Bacteriophage, Phage group, Bacteriophages, MESH: Genetic Variation, MESH : Viral Proteins, 2. Zero hunger, Genetics, biology, Dairy Fermentation, MESH: Genomics, Genomics, MESH : Host Specificity, MESH : Bacteriophages, Lactococcus lactis, [ SDV.BBM.GTP ] Life Sciences [q-bio]/Biochemistry, Molecular Biology/Genomics [q-bio.GN], MESH: Calcium, [SDV.MP.VIR]Life Sciences [q-bio]/Microbiology and Parasitology/Virology, DNA microarray, Research Article, Biotechnology, MESH: Host Specificity, Context (language use), [ SDV.MP.VIR ] Life Sciences [q-bio]/Microbiology and Parasitology/Virology, Host Specificity, Microbiology, Dairy, Viral Proteins, 03 medical and health sciences, MESH : Genetic Variation, [SDV.BBM.GTP]Life Sciences [q-bio]/Biochemistry, Molecular Biology/Genomics [q-bio.GN], MESH: Bacteriophages, MESH : Calcium, Host (biology), MESH : Genomics, Genetic Variation, biology.organism_classification, MESH: Viral Proteins, [SDV.MP.BAC]Life Sciences [q-bio]/Microbiology and Parasitology/Bacteriology, 030104 developmental biology, MESH: Lactococcus lactis, Fermentation, Receptor-binding protein, Calcium, MESH : Lactococcus lactis
Abstract

Background Despite continuous research efforts, bacterio(phages) infecting Lactococcus lactis starter strains persist as a major threat to dairy fermentations. The lactococcal P335 phages, which are currently classified into four sub-groups (I-IV), are the second most frequently isolated phage group in an industrial dairy context. Results The current work describes the isolation and comparative genomic analysis of 17 novel P335 group phages. Detailed analysis of the genomic region of P335 phages encoding the so-called “baseplate”, which includes the receptor binding protein (RBP) was combined with a functional characterization of the RBP of sub-group III and IV phages. Additionally, calcium-dependence assays revealed a specific requirement for calcium by sub-group IV phages while host range analysis highlighted a higher number of strains with CWPS type A (11 of 39 strains) are infected by the P335 phages assessed in this study than those with a C (five strains), B (three of 39 strains) or unknown (one of 39 strains) CWPS type. Conclusions These analyses revealed significant divergence among RBP sequences, apparently reflecting their unique interactions with the host and particularly for strains with a type A CWPS. The implications of the genomic architecture of lactococcal P335 phages on serving as a general model for Siphoviridae phages are discussed. Electronic supplementary material The online version of this article (doi:10.1186/s12864-017-3537-5) contains supplementary material, which is available to authorized users.

Published
2017
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2. Characterization of Unique Signature Sequences in the Divergent Maternal Protein Bcl2l10

Author

Germain Gillet, Abdel Aouacheria, François Penin, Aurélie Cornut-Thibaut, Yannis Guillemin, Institut de biologie et chimie des protéines [Lyon] (IBCP), Université Claude Bernard Lyon 1 (UCBL), Université de Lyon-Université de Lyon-Centre National de la Recherche Scientifique (CNRS), Centre de Recherche en Cancérologie de Lyon (UNICANCER/CRCL), Centre Léon Bérard [Lyon]-Université Claude Bernard Lyon 1 (UCBL), Université de Lyon-Université de Lyon-Centre National de la Recherche Scientifique (CNRS)-Institut National de la Santé et de la Recherche Médicale (INSERM), Laboratoire de Biologie Moléculaire de la Cellule (LBMC), École normale supérieure - Lyon (ENS Lyon)-Université Claude Bernard Lyon 1 (UCBL), Université de Lyon-Université de Lyon-Institut National de la Santé et de la Recherche Médicale (INSERM)-Centre National de la Recherche Scientifique (CNRS), Ganivet, Agnès, Institut de biologie et chimie des protéines [Lyon] ( IBCP ), Université Claude Bernard Lyon 1 ( UCBL ), Université de Lyon-Université de Lyon-Centre National de la Recherche Scientifique ( CNRS ), Centre de Recherche en Cancérologie de Lyon ( CRCL ), Université de Lyon-Université de Lyon-Centre Léon Bérard [Lyon]-Institut National de la Santé et de la Recherche Médicale ( INSERM ) -Centre National de la Recherche Scientifique ( CNRS ), Laboratoire de Biologie Moléculaire de la Cellule ( LBMC ), École normale supérieure - Lyon ( ENS Lyon ) -Université Claude Bernard Lyon 1 ( UCBL ), Université de Lyon-Université de Lyon-Institut National de la Santé et de la Recherche Médicale ( INSERM ) -Centre National de la Recherche Scientifique ( CNRS ), and École normale supérieure de Lyon (ENS de Lyon)-Université Claude Bernard Lyon 1 (UCBL)

Subjects
MESH : Cell Line, MESH : Molecular Sequence Data, Amino Acid Motifs, MESH: Amino Acid Sequence, MESH: Base Sequence, MESH : Proto-Oncogene Proteins c-bcl-2, [ SDV.CAN ] Life Sciences [q-bio]/Cancer, MESH: INDEL Mutation, protein sequence, MESH: Amino Acid Motifs, MESH: Protein Structure, Tertiary, 0302 clinical medicine, INDEL Mutation, Databases, Genetic, genetic polymorphism, MESH : Female, MESH: Animals, MESH: Genetic Variation, [SDV.BDD]Life Sciences [q-bio]/Development Biology, Peptide sequence, MESH: Databases, Genetic, MESH: Evolution, Molecular, Sequence Deletion, Genetics, chemistry.chemical_classification, 0303 health sciences, MESH : Amino Acid Sequence, MESH : Sequence Alignment, apoptosis, Vertebrate, MESH: Sequence Deletion, Amino acid, Proto-Oncogene Proteins c-bcl-2, MESH: Calcium, 030220 oncology & carcinogenesis, Female, MESH : Protein Structure, Tertiary, MESH : Amino Acid Motifs, Molecular Sequence Data, MESH: Sequence Alignment, [SDV.CAN]Life Sciences [q-bio]/Cancer, Sequence alignment, Biology, Cell Line, MESH: Oocytes, Evolution, Molecular, 03 medical and health sciences, Bcl-2 family, MESH : Genetic Variation, biology.animal, evolution, [SDV.BDD] Life Sciences [q-bio]/Development Biology, Genetic variation, Homologous chromosome, Animals, Humans, MESH : HeLa Cells, MESH : Evolution, Molecular, Amino Acid Sequence, MESH : Calcium, MESH : Databases, Genetic, Indel, MESH : INDEL Mutation, Molecular Biology, Ecology, Evolution, Behavior and Systematics, 030304 developmental biology, MESH : Mutagenesis, Insertional, calcium, MESH: Humans, MESH: Molecular Sequence Data, Base Sequence, MESH : Sequence Deletion, MESH: Apoptosis, MESH : Humans, MESH : Oocytes, Genetic Variation, Protein Structure, Tertiary, MESH: Cell Line, Mutagenesis, Insertional, MESH: Mutagenesis, Insertional, MESH: Proto-Oncogene Proteins c-bcl-2, chemistry, MESH: HeLa Cells, Oocytes, MESH : Base Sequence, MESH : Animals, Sequence Alignment, MESH: Female, MESH : Apoptosis, HeLa Cells
Abstract

International audience; Insertions or deletions (indels) of amino acids residues have been recognized as an important source of genetic and structural divergence between paralogous Bcl-2 family members. However, these signature sequences have not so far been extensively investigated amongst orthologous Bcl-2 family proteins. Bcl2l10 is an antiapoptotic member of the Bcl-2 family that has evolved rapidly throughout the vertebrate lineage and which shows conserved abundant expression in eggs and oocytes. In this paper, we have unraveled two major sites of divergence between human Bcl2l10 and its vertebrate homologs. The first one provides length variation at the N-terminus (before the BH4 domain) and the second one is located between the predicted α5-α6 pore-forming helices, providing an unprecedented case in the superfamily of helix-bundled pore-forming proteins. These two particular indels were studied phylogenetically and through biochemical and cell biological techniques, including truncation and site-directed mutagenesis. While deletion of the N-terminal extension had no significant functional impact in HeLa cells, our results suggest that the human Bcl2l10 protein evolved a calcium-binding motif in its α5-α6 interhelical region by acquiring critical negatively charged residues. Considering the reliance of female eggs on calcium-dependent proteins and calcium-regulated processes and the exceptional longevity of oocytes in the primate lineage, we propose that this microstructural variation may be an adaptive feature associated with high maternal expression of this Bcl-2 family member.

Published
2011
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3. Epsilon toxin from Clostridium perfringens acts on oligodendrocytes without forming pores, and causes demyelination

Author

Flavia Heid, Serge Pauillac, Frédéric Doussau, Bernard Poulain, Jean-Luc Dupont, Stéphane Gaillard, Jean-Louis Bossu, Laetitia Wioland, Philippe Isope, Michel R. Popoff, Institut des Neurosciences Cellulaires et Intégratives ( INCI ), Université Louis Pasteur - Strasbourg I-Centre National de la Recherche Scientifique ( CNRS ), Bactéries anaérobies et Toxines, Institut Pasteur [Paris], supported by a doctoral grant from the Mission pour la Recherche et I’Innovation Scientifique – Délégation Générale à I’Armement (MRIS/DGA) for L.W., and by CNRS funding for B.P, Institut des Neurosciences Cellulaires et Intégratives (INCI), Université Louis Pasteur - Strasbourg I-Centre National de la Recherche Scientifique (CNRS), and Institut Pasteur [Paris] (IP)

Subjects
MESH : Clostridium perfringens, Cell type, MESH: Demyelinating Diseases, MESH: Rats, MESH : Cerebellum, Clostridium perfringens, Immunology, Purkinje cell, [ SDV.TOX ] Life Sciences [q-bio]/Toxicology, Bacterial Toxins, Glutamic Acid, Biology, medicine.disease_cause, Microbiology, MESH : Glutamic Acid, Virology, Cerebellum, MESH : Cells, Cultured, medicine, Animals, MESH: Animals, Receptor, MESH : Calcium, Cells, Cultured, MESH: Clostridium perfringens, MESH : Rats, MESH: Oligodendroglia, MESH: Glutamic Acid, MESH : Oligodendroglia, MESH: Cerebellum, Cell biology, Rats, MESH : Demyelinating Diseases, Oligodendroglia, medicine.anatomical_structure, MESH: Bacterial Toxins, Biochemistry, Metabotropic glutamate receptor, MESH: Calcium, [SDV.TOX]Life Sciences [q-bio]/Toxicology, MESH : Bacterial Toxins, NMDA receptor, Metabotropic glutamate receptor 1, Calcium, MESH : Animals, Intracellular, MESH: Cells, Cultured, Demyelinating Diseases
Abstract

International audience; Epsilon toxin (ET) is produced by Clostridium perfringens types B and D and causes severe neurological disorders in animals. ET has been observed binding to white matter, suggesting that it may target oligodendrocytes. In primary cultures containing oligodendrocytes and astrocytes, we found that ET (10(-9) M and 10(-7) M) binds to oligodendrocytes, but not to astrocytes. ET induces an increase in extracellular glutamate, and produces oscillations of intracellular Ca(2+) concentration in oligodendrocytes. These effects occurred without any change in the transmembrane resistance of oligodendrocytes, underlining that ET acts through a pore-independent mechanism. Pharmacological investigations revealed that the Ca(2+) oscillations are caused by the ET-induced rise in extracellular glutamate concentration. Indeed, the blockade of metabotropic glutamate receptors type 1 (mGluR1) prevented ET-induced Ca(2+) signals. Activation of the N-methyl-D-aspartate receptor (NMDA-R) is also involved, but to a lesser extent. Oligodendrocytes are responsible for myelinating neuronal axons. Using organotypic cultures of cerebellar slices, we found that ET induced the demyelination of Purkinje cell axons within 24 h. As this effect was suppressed by antagonizing mGluR1 and NMDA-R, demyelination is therefore caused by the initial ET-induced rise in extracellular glutamate concentration. This study reveals the novel possibility that ET can act on oligodendrocytes, thereby causing demyelination. Moreover, it suggests that for certain cell types such as oligodendrocytes, ET can act without forming pores, namely through the activation of an undefined receptor-mediated pathway.

Published
2014
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4. Dendrochemical patterns of calcium, zinc, and potassium related to internal factors detected by energy dispersive X-ray fluorescence (EDXRF)

Author

Håkan Grudd, Don A. Vroblesky, Michel Chalot, Jean Christophe Balouet, François Beaujard, Kevin T. Smith, Walter C. Shortle, Joel G. Burken, United States Department of Agriculture, International Environmental, Université de Franche-Comté (UFC), Université de Lorraine (UL), Laboratoire de Physique et Physiologie Intégratives de l'Arbre Fruitier et Forestier (PIAF), Institut National de la Recherche Agronomique (INRA)-Université Blaise Pascal - Clermont-Ferrand 2 (UBP), Bolin Centre for Climate Research, Stockholm University, United States Geological Survey [Reston] (USGS), Missouri University of Science and Technology (Missouri S&T), University of Missouri System, Laboratoire Chrono-environnement - UFC (UMR 6249) (LCE), Centre National de la Recherche Scientifique (CNRS)-Université de Franche-Comté (UFC)-Université Bourgogne Franche-Comté [COMUE] (UBFC), Laboratoire Chrono-environnement - CNRS - UBFC (UMR 6249) (LCE), Centre National de la Recherche Scientifique (CNRS)-Université de Franche-Comté (UFC), Université Bourgogne Franche-Comté [COMUE] (UBFC)-Université Bourgogne Franche-Comté [COMUE] (UBFC), Laboratoire Chrono-environnement ( LCE ), Université Bourgogne Franche-Comté ( UBFC ) -Université de Franche-Comté ( UFC ) -Centre National de la Recherche Scientifique ( CNRS ), Laboratoire de Physique et Physiologie Intégratives de l'Arbre Fruitier et Forestier ( PIAF ), Institut National de la Recherche Agronomique ( INRA ) -Université Blaise Pascal - Clermont-Ferrand 2 ( UBP ), Laboratoire Chrono-environnement (UMR 6249) (LCE), and Université Bourgogne Franche-Comté [COMUE] (UBFC)

Subjects
bois, Wood calcium, Health, Toxicology and Mutagenesis, Potassium, [SDV]Life Sciences [q-bio], Cation distribution, MESH: Zinc, Annual growth %, Trees, MESH : Trees, MESH : Environmental Monitoring, [SDV.EE]Life Sciences [q-bio]/Ecology, environment, MESH: Spectrometry, X-Ray Emission, General Medicine, Contamination, Pollution, Zinc, Metals, Environmental chemistry, MESH: Calcium, Dendrochemistry, Wood potassium, MESH: Environmental Monitoring, Environmental Monitoring, Environmental Engineering, chemistry.chemical_element, X-ray fluorescence, Calcium, dendrochimie, [ SDV.EE ] Life Sciences [q-bio]/Ecology, environment, MESH : Spectrometry, X-Ray Emission, Environmental Chemistry, MESH : Calcium, MESH: Metals, MESH : Metals, Public Health, Environmental and Occupational Health, Spectrometry, X-Ray Emission, General Chemistry, MESH : Zinc, Highly sensitive, MESH: Trees, MESH : Potassium, chemistry, 13. Climate action, MESH: Potassium, Biomineralization
Abstract

International audience; Energy dispersive X-ray fluorescence (EDXRF) provides highly sensitive and precise spatial resolution of cation content in individual annual growth rings in trees. The sensitivity and precision have prompted successful applications to forensic dendrochemistry and the timing of environmental releases of contaminants. These applications have highlighted the need to distinguish dendrochemical effects of internal processes from environmental contamination. Calcium, potassium, and zinc are three marker cations that illustrate the influence of these processes. We found changes in cation chemistry in tree rings potentially due to biomineralization, development of cracks or checks, heartwood/sapwood differentiation, intra-annual processes, and compartmentalization of infection. Distinguishing internal from external processes that affect dendrochemistry will enhance the value of EDXRF for both physiological and forensic investigations.

Published
2014
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5. Synthesis and in vitro characterisation of ifenprodil-based fluorescein conjugates as GluN1/GluN2B N-Methyl-D-aspartate receptor antagonists

Author

Louisa Barré, Olivier Nicole, Eric T. MacKenzie, Alain Buisson, Cécile Perrio, Javier Becerril-Ortega, Nathalie Colloc'h, Martine Dhilly, Centre-Imagerie, Neurosciences, et Application aux Pathologies (CI-NAPS - UMR 6232), Université de Caen Normandie (UNICAEN), Normandie Université (NU)-Normandie Université (NU)-Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Université Paris Descartes - Paris 5 (UPD5)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Centre National de la Recherche Scientifique (CNRS), NICOLE, Olivier, 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 ), Imagerie et Stratégies Thérapeutiques des pathologies Cérébrales et Tumorales (ISTCT), Normandie Université (NU)-Normandie Université (NU)-Centre National de la Recherche Scientifique (CNRS)-Commissariat à l'énergie atomique et aux énergies alternatives (CEA), Centre-Imagerie, Neurosciences, et Application aux Pathologies ( CI-NAPS - UMR 6232 ), and Normandie Université ( NU ) -Normandie Université ( NU ) -Commissariat à l'énergie atomique et aux énergies alternatives ( CEA ) -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, Models, Molecular, MESH : Fluorescein, MESH : Piperidines, [SDV]Life Sciences [q-bio], [SDV.NEU.NB]Life Sciences [q-bio]/Neurons and Cognition [q-bio.NC]/Neurobiology, MESH: Neurons, Biochemistry, chemistry.chemical_compound, Mice, Piperidines, MESH : Neuroprotective Agents, Fluorescein, MESH: Fluorescein, Receptor, Cells, Cultured, ComputingMilieux_MISCELLANEOUS, Neurons, Brain, MESH: Neuroprotective Agents, Neuroprotective Agents, MESH: Piperidines, [ SDV.NEU.NB ] Life Sciences [q-bio]/Neurons and Cognition [q-bio.NC]/Neurobiology, MESH: Calcium, MESH: HEK293 Cells, Molecular Medicine, NMDA receptor, [CHIM.RADIO]Chemical Sciences/Radiochemistry, [ CHIM.RADIO ] Chemical Sciences/Radiochemistry, Fluorophore, N-Methylaspartate, Stereochemistry, Neuroprotection, Receptors, N-Methyl-D-Aspartate, MESH : Neurons, Calcium imaging, Ifenprodil, Animals, Humans, Binding site, MESH : Calcium, Molecular Biology, MESH: Receptors, N-Methyl-D-Aspartate, Binding Sites, [ SDV ] Life Sciences [q-bio], Organic Chemistry, [SDV.NEU.NB] Life Sciences [q-bio]/Neurons and Cognition [q-bio.NC]/Neurobiology, MESH : HEK293 Cells, Radiography, HEK293 Cells, chemistry, MESH: Binding Sites, Calcium, MESH : Receptors, N-Methyl-D-Aspartate, MESH : Binding Sites
Abstract

International audience; GluN2B-containing NMDA receptors are involved in many important physiological functions and play a pivotal role in mediating pain as well as in several neurodegenerative disorders. We aimed to develop fluorescent probes to target the GluN2B subunit selectively in order to allow better understanding of the relationships between receptor localisation and physiological importance. Ifenprodil, known as the GluNR2B antagonist of reference, was chosen as the template for the elaboration of probes. We had previously reported a fluorescein conjugate that was shown (by confocal microscopy imaging of DS-red-labelled cortical neurons) to bind specifically to GluN2B. To elaborate this probe, we explored the influence of both the nature and the attachment point of the spacer between the fluorophore and the parent compound, ifenprodil. We performed chemical modifications of ifenprodil at the benzylic position and on the phenol ring by introducing secondary amine or amide functions and evaluated alkyl chains from two to 20 bonds either including or not including secondary amide functions as spacers. The previously developed probe was found to display the greatest activity in the inhibition of NMDA-induced Ca(2+) influx by calcium imaging experiments on HEK293 cells transfected with the cDNA encoding for GluN1-1A and GluN2B. Further investigations revealed that this probe had a neuroprotective effect equivalent to that of ifenprodil in a standard test for neurotoxicity. Despite effects of lesser amplitude with these probes relative to ifenprodil, we demonstrated that they displaced [(3) H]ifenprodil in mouse brain slices in a similar manner.

Published
2013
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6. NMR studies of the interaction of calmodulin with IQ motif peptides

Author

Walter J. Chazin, Benjamin Chagot, Steven M. Damo, Michael D. Feldkamp, Center for Structural Biology, Vanderbilt University [Nashville], Ingénierie Moléculaire et Physiopathologie Articulaire ( IMoPA ), Université de Lorraine ( UL ) -Centre National de la Recherche Scientifique ( CNRS ), Ingénierie Moléculaire et Physiopathologie Articulaire (IMoPA), Université de Lorraine (UL)-Centre National de la Recherche Scientifique (CNRS), and Centre National de la Recherche Scientifique (CNRS)-Université de Lorraine (UL)

Subjects
MESH: Signal Transduction, MESH : NAV1.5 Voltage-Gated Sodium Channel, Amino Acid Motifs, Intracellular Space, MESH : Intracellular Space, Peptide, Plasma protein binding, EF-hand, Na 15, 15 N-1 H HSQC, IQ motif, NAV1.5 Voltage-Gated Sodium Channel, MESH: Amino Acid Motifs, MESH: Apoproteins, Calcium-binding protein, MESH: Nuclear Magnetic Resonance, Biomolecular, MESH: Peptide Fragments, Calcium signaling, chemistry.chemical_classification, 0303 health sciences, [SDV.BBM.BS]Life Sciences [q-bio]/Biochemistry, Molecular Biology/Structural Biology [q-bio.BM], biology, EF hand, 030302 biochemistry & molecular biology, MESH : Apoproteins, MESH : Protein Binding, Nuclear magnetic resonance spectroscopy, MESH: Calmodulin, 3. Good health, [SDV.BBM.BP]Life Sciences [q-bio]/Biochemistry, Molecular Biology/Biophysics, Biochemistry, MESH: Calcium, MESH: Intracellular Space, Protein Binding, Signal Transduction, MESH : Calmodulin, [ SDV.BBM.BP ] Life Sciences [q-bio]/Biochemistry, Molecular Biology/Biophysics, MESH : Amino Acid Motifs, animal structures, Calmodulin, MESH : Nuclear Magnetic Resonance, Biomolecular, calcium signaling, Article, 03 medical and health sciences, NMR spectroscopy, MESH: Protein Binding, Humans, MESH : Calcium, Nuclear Magnetic Resonance, Biomolecular, Ion channel, 030304 developmental biology, MESH : Signal Transduction, MESH: Humans, protein complex, MESH : Peptide Fragments, MESH : Humans, MESH: NAV1.5 Voltage-Gated Sodium Channel, calcium binding protein, Peptide Fragments, co-expression, chemistry, biology.protein, Calcium, Apoproteins, [ SDV.BBM.BS ] Life Sciences [q-bio]/Biochemistry, Molecular Biology/Biomolecules [q-bio.BM]
Abstract

International audience; Calmodulin (CaM) is a ubiquitous EF-hand calcium sensor protein that transduces calcium signals in a wide range of signaling pathways. Structural analysis of complexes with peptides has provided valuable insights into the remarkable variety in the way in which CaM interacts with and activates its targets. Among these various targets, CaM has been shown to be an essential component of a calcium-sensing regulatory apparatus for a number of voltage-gated ion channels. NMR spectroscopy has proven to be a powerful tool for the structural characterization of CaM-peptide complexes, in particular for the study of IQ motifs, which bind CaM at the basal level of calcium in cells and thereby serve to localize CaM to its sites of action. We describe here methods for the robust expression and purification of CaM isotopically enriched for NMR analysis, as well as for the complex of CaM with a peptide derived from the IQ motif sequence of the human cardiac sodium channel Na(V)1.5. We also describe methods for NMR analysis of titrations of CaM with IQ motif peptides to determine the stoichiometry of the complex and to identify the residues at the binding interface.

Published
2013
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7. Glycogen synthase kinase 3-mediated voltage-dependent anion channel phosphorylation controls outer mitochondrial membrane permeability during lipid accumulation

Author

Guido Kroemer, Cécile Martel, Céline Henry, Catherine Brenner, Elena Fanelli, Maya Allouche, Joël Chopineau, Giuseppe Calamita, Davide Degli Esposti, Antoinette Lemoine, Céline Boursier, Université Joseph Fourier - Grenoble 1 (UJF), Institut Charles Gerhardt Montpellier - Institut de Chimie Moléculaire et des Matériaux de Montpellier (ICGM ICMMM), Université Montpellier 1 (UM1)-Université Montpellier 2 - Sciences et Techniques (UM2)-Ecole Nationale Supérieure de Chimie de Montpellier (ENSCM)-Institut de Chimie du CNRS (INC)-Centre National de la Recherche Scientifique (CNRS), Réponses cellulaires au microenvironnement et cancer, Institut National de la Santé et de la Recherche Médicale (INSERM), Università degli studi di Bari Aldo Moro = University of Bari Aldo Moro (UNIBA), Protéines et Nanotechnologies en Sciences Séparatives, Université Paris-Sud - Paris 11 (UP11), Centre Ressources Autisme de Bourgogne (CHU de Dijon) (CRAB), Centre Hospitalier Universitaire de Dijon - Hôpital François Mitterrand (CHU Dijon), Université de Nîmes (UNIMES), Centre de Recherche des Cordeliers (CRC (UMR_S 872)), Université Pierre et Marie Curie - Paris 6 (UPMC)-Université Paris Descartes - Paris 5 (UPD5)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Centre National de la Recherche Scientifique (CNRS), Apoptose, cancer et immunité (U848), Université Paris-Sud - Paris 11 (UP11)-Institut Gustave Roussy (IGR)-Institut National de la Santé et de la Recherche Médicale (INSERM), Plateforme de métabolomique, Direction de la recherche [Gustave Roussy], Institut Gustave Roussy (IGR)-Institut Gustave Roussy (IGR), Pôle de biologie, Assistance publique - Hôpitaux de Paris (AP-HP) (AP-HP)-Hôpital Européen Georges Pompidou [APHP] (HEGP), Assistance publique - Hôpitaux de Paris (AP-HP) (AP-HP)-Hôpitaux Universitaires Paris Ouest - Hôpitaux Universitaires Île de France Ouest (HUPO)-Hôpitaux Universitaires Paris Ouest - Hôpitaux Universitaires Île de France Ouest (HUPO), Université Paris Descartes - Faculté de Médecine (UPD5 Médecine), Université Paris Descartes - Paris 5 (UPD5), Microenvironnement et Physiopathologie de la Differenciation, Université Paris-Sud - Paris 11 (UP11)-Institut National de la Santé et de la Recherche Médicale (INSERM), Laboratoire d'Excellence en Recherche sur le Médicament et l'Innovation Thérapeutique [Châtenay-Malabry] (LabEx LERMIT), University of Paris Sud, Institut National pour le Cancer (INCa) [2008-1-PL BIO-04-CNRS ON1], LabEx LERMIT, PRES UniverSud Paris, Association pour la Recherche sur le Cancer, ANR [ANR-08PCVI-0008-01], MIUR [PRIN20089SRS2X_003], Fondazione Cassa di Risparmio di Puglia, Chopineau, Joel, Université Joseph Fourier - Grenoble 1 ( UJF ), Institut Charles Gerhardt Montpellier - Institut de Chimie Moléculaire et des Matériaux de Montpellier ( ICGM ICMMM ), Université Montpellier 1 ( UM1 ) -Université Montpellier 2 - Sciences et Techniques ( UM2 ) -Ecole Nationale Supérieure de Chimie de Montpellier ( ENSCM ) -Université de Montpellier ( UM ) -Centre National de la Recherche Scientifique ( CNRS ), Institut National de la Santé et de la Recherche Médicale ( INSERM ), University of Bari Aldo Moro ( UNIBA ), Université Paris-Sud - Paris 11 ( UP11 ), Centre Ressources Autisme de Bourgogne (CHU de Dijon) ( CRAB ), Centre Hospitalier Universitaire de Dijon - Hôpital François Mitterrand ( CHU Dijon ), Université de Nîmes ( UNIMES ), Centre de Recherche des Cordeliers ( CRC (UMR_S 872) ), Université Pierre et Marie Curie - Paris 6 ( UPMC ) -Université Paris Descartes - Paris 5 ( UPD5 ) -Institut National de la Santé et de la Recherche Médicale ( INSERM ) -Centre National de la Recherche Scientifique ( CNRS ), Apoptose, cancer et immunité ( U848 ), Université Paris-Sud - Paris 11 ( UP11 ) -Institut Gustave Roussy ( IGR ) -Institut National de la Santé et de la Recherche Médicale ( INSERM ), Institut Gustave Roussy ( IGR ) -Institut Gustave Roussy ( IGR ), Assistance publique - Hôpitaux de Paris (AP-HP)-Hôpital Européen Georges Pompidou [APHP] ( HEGP ), Université Paris Descartes - Faculté de Médecine ( UPD5 Médecine ), Université Paris Descartes - Paris 5 ( UPD5 ), Université Paris-Sud - Paris 11 ( UP11 ) -Institut National de la Santé et de la Recherche Médicale ( INSERM ), PRES UniverSud ( CNRS UMR 8159 ), Université de Versailles Saint-Quentin-en-Yvelines ( UVSQ ), Ecole Nationale Supérieure de Chimie de Montpellier (ENSCM)-Centre National de la Recherche Scientifique (CNRS)-Université de Montpellier (UM)-Université Montpellier 1 (UM1)-Université Montpellier 2 - Sciences et Techniques (UM2)-Institut de Chimie du CNRS (INC), University of Bari Aldo Moro (UNIBA), and Institut National de la Santé et de la Recherche Médicale (INSERM)-Institut Gustave Roussy (IGR)-Université Paris-Sud - Paris 11 (UP11)

Subjects
Male, MESH : Fatty Liver, MESH : Hepatocytes, Mitochondrion, ACTIVATION, MESH: Hepatocytes, Glycogen Synthase Kinase 3, Mice, MESH : Phosphorylation, 0302 clinical medicine, MESH: Mitochondrial Membranes, MESH : Lipid Metabolism, GSK-3, Voltage-Dependent Anion Channels, MESH : Female, MESH: Animals, Phosphorylation, MESH: Fatty Liver, Cells, Cultured, MESH: Glycogen Synthase Kinase 3, MESH: Lipid Metabolism, 0303 health sciences, INSULIN-RESISTANCE, biology, VDAC, MESH : Voltage-Dependent Anion Channels, NONALCOHOLIC STEATOHEPATITIS, Fatty liver, 3. Good health, Cell biology, APOPTOSIS, [SDV.BBM.BP]Life Sciences [q-bio]/Biochemistry, Molecular Biology/Biophysics, Lipotoxicity, Biochemistry, 030220 oncology & carcinogenesis, MESH: Calcium, Mitochondrial Membranes, Female, MESH: Cells, Cultured, [ SDV.BBM.BP ] Life Sciences [q-bio]/Biochemistry, Molecular Biology/Biophysics, Voltage-dependent anion channel, MESH : Glycogen Synthase Kinase 3, MESH : Male, bcl-X Protein, INHIBITION, [SDV.BBM.BP] Life Sciences [q-bio]/Biochemistry, Molecular Biology/Biophysics, MESH : Mice, Inbred C57BL, macromolecular substances, MESH: bcl-X Protein, 03 medical and health sciences, MESH : Mitochondrial Membranes, MESH: Mice, Inbred C57BL, LIVER STEATOSIS, MESH : Mice, MESH : Cells, Cultured, medicine, Animals, Humans, TRANSITION PORE, MODULATION, MESH : Calcium, MESH: Mice, 030304 developmental biology, MESH: Humans, Hepatology, MESH: Phosphorylation, MESH : bcl-X Protein, MESH : Humans, MESH: Voltage-Dependent Anion Channels, Lipid metabolism, Lipid Metabolism, medicine.disease, MESH: Male, Fatty Liver, Mice, Inbred C57BL, CELL-DEATH, Hepatocytes, biology.protein, Calcium, MESH : Animals, Steatosis, MESH: Female
Abstract

International audience; UNLABELLED: Nonalcoholic steatosis is a liver pathology characterized by fat accumulation and severe metabolic alterations involving early mitochondrial impairment and late hepatocyte cell death. However, mitochondrial dysfunction mechanisms remain elusive. Using four models of nonalcoholic steatosis, i.e., livers from patients with fatty liver disease, ob/ob mice, mice fed a high-fat diet, and in vitro models of lipotoxicity, we show that outer mitochondrial membrane permeability is altered and identified a posttranslational modification of voltage-dependent anion channel (VDAC), a membrane channel and NADH oxidase, as a cause of early mitochondrial dysfunction. Thus, in nonalcoholic steatosis VDAC exhibits reduced threonine phosphorylation, which increases the influx of water and calcium into mitochondria, sensitizes the organelle to matrix swelling, depolarization, and cytochrome c release without inducing cell death. This also amplifies VDAC enzymatic and channel activities regulation by calcium and modifies its interaction with proteic partners. Moreover, lipid accumulation triggers a rapid lack of VDAC phosphorylation by glycogen synthase kinase 3 (GSK3). Pharmacological and genetic manipulations proved GSK3 to be responsible for VDAC phosphorylation in normal cells. Notably, VDAC phosphorylation level correlated with steatosis severity in patients. CONCLUSION: VDAC acts as an early sensor of lipid toxicity and its GSK3-mediated phosphorylation status controls outer mitochondrial membrane permeabilization in hepatosteatosis.

Published
2013
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8. Glycosaminoglycan mimetics trigger IP3-dependent intracellular calcium release in myoblasts

Author

Isabelle Guillet-Deniau, Dulce Papy-Garcia, Isabelle Martelly, José Courty, Guy Raymond, Dominique Singabraya, Aurélie Vandebrouck, Christian Cognard, Bruno Constantin, Croissance cellulaire, réparation et régénération tissulaires (CRRET), Université Paris-Est Créteil Val-de-Marne - Paris 12 (UPEC UP12)-Centre National de la Recherche Scientifique (CNRS), Institut de physiologie et biologie cellulaires (IPBC), Université de Poitiers-Centre National de la Recherche Scientifique (CNRS), Institut Cochin (UMR_S567 / UMR 8104), 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 de Physiologie et Biologie Cellulaires (IPBC), Institut Cochin (IC UM3 (UMR 8104 / U1016)), Université Paris Descartes - Paris 5 (UPD5)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Centre National de la Recherche Scientifique (CNRS), Laboratoire CRRET, EAC/CNRS-7149, Université Paris-Est Créteil Val-de-Marne - Paris 12 (UPEC UP12), Laboratoire de recherche sur la croissance cellulaire, la réparation et la régénération tissulaires ( CRRET ), Université Paris-Est Créteil Val-de-Marne - Paris 12 ( UPEC UP12 ) -Centre National de la Recherche Scientifique ( CNRS ), Institut de physiologie et biologie cellulaires ( IPBC ), Université de Poitiers-Centre National de la Recherche Scientifique ( CNRS ), Institut Cochin ( UMR_S567 / UMR 8104 ), and 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
MESH : Calcium, Dietary, MESH : Cell Line, MESH: Signal Transduction, Cytoplasm, MESH: Heparin, [SDV]Life Sciences [q-bio], Cellular homeostasis, [SDV.BC.BC]Life Sciences [q-bio]/Cellular Biology/Subcellular Processes [q-bio.SC], Muscle Development, Calcium in biology, MESH: Ryanodine Receptor Calcium Release Channel, Myoblasts, Mice, 0302 clinical medicine, MESH : Receptor, Serotonin, 5-HT2A, [SDV.BC.IC]Life Sciences [q-bio]/Cellular Biology/Cell Behavior [q-bio.CB], Receptor, Serotonin, 5-HT2A, MESH: Animals, MESH : Ryanodine Receptor Calcium Release Channel, Glycosaminoglycans, 0303 health sciences, MESH : Rats, Ryanodine receptor, Myogenesis, MESH: Regeneration, MESH: Satellite Cells, Skeletal Muscle, Cell Differentiation, MESH: Glycosaminoglycans, Cell biology, [SDV.BBM.BC]Life Sciences [q-bio]/Biochemistry, Molecular Biology/Biomolecules [q-bio.BM], Biochemistry, MESH: Calcium, MESH: Muscle Development, MESH : Cell Differentiation, MESH: Calcium, Dietary, Intracellular, Signal Transduction, MESH: Cell Differentiation, MESH : Cytoplasm, Satellite Cells, Skeletal Muscle, MESH: Rats, Inositol Phosphates, MESH : Muscle Development, MESH : Inositol Phosphates, chemistry.chemical_element, MESH : Regeneration, [SDV.BC]Life Sciences [q-bio]/Cellular Biology, Biology, Calcium, MESH : Rats, Wistar, MESH : Myoblasts, Cell Line, 03 medical and health sciences, MESH : Satellite Cells, Skeletal Muscle, MESH : Mice, MESH : Glycosaminoglycans, [SDV.MHEP.PHY]Life Sciences [q-bio]/Human health and pathology/Tissues and Organs [q-bio.TO], Animals, Regeneration, [SDV.BBM]Life Sciences [q-bio]/Biochemistry, Molecular Biology, MESH: Myoblasts, Rats, Wistar, MESH : Calcium, Molecular Biology, MESH: Mice, 030304 developmental biology, MESH : Signal Transduction, Calcium metabolism, Heparin, MESH: Cytoplasm, MESH: Receptor, Serotonin, 5-HT2A, MESH: Clone Cells, [ SDV.BC.BC ] Life Sciences [q-bio]/Cellular Biology/Subcellular Processes [q-bio.SC], MESH : Clone Cells, Ryanodine Receptor Calcium Release Channel, MESH: Rats, Wistar, Inositol trisphosphate receptor, MESH: Inositol Phosphates, Clone Cells, Rats, MESH: Cell Line, Calcium, Dietary, MESH : Heparin, chemistry, MESH : Animals, 030217 neurology & neurosurgery
Abstract

International audience; Glycosaminoglycans (GAG) are sulfated polysaccharides that play an important role in regulating cell functions. GAG mimetics called RGTAs (for ReGeneraTing Agents) have been shown to stimulate tissue repair. In particular they accelerate myogenesis, in part via their heparin-mimetic property towards growth factors. RGTAs also increase activity of calcium-dependent intracellular protease suggesting an effect on calcium cellular homeostasis. This effect was presently investigated on myoblasts in vitro using one member of the RGTA family molecule named OTR4120. We have shown that OTR4120 or heparin induced transient increases of intracellular calcium concentration ([Ca(2+)]i) in pre-fusing myoblasts from both mouse SolD7 cell line and rat skeletal muscle satellite cells grown in primary culture by mobilising sarcoplasmic reticulum store. This [Ca(2+)]i was not mediated by ryanodine receptors but instead resulted from stimulation of the Inositol-3 phosphate-phospholipase C activation pathway. OTR4120-induced calcium transient was not mediated through an ATP, nor a tyrosine kinase, nor an acetylcholine receptor but principally through serotonin 5-HT2A receptor. This original finding shows that the GAG mimetic can induce calcium signal through serotonin receptors and the IP3 pathway may be relevant to its ability to favour myoblast differentiation. It supports a novel and unexpected function of GAGs in the regulation of calcium homeostasis.

Published
2010
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9. Bacterial Toxins and the Nervous System: Neurotoxins and Multipotential Toxins Interacting with Neuronal Cells

Author

Bernard Poulain, Michel R. Popoff, Bactéries anaérobies et Toxines, Institut Pasteur [Paris] (IP), Neurotransmission et sécrétion neuroendocrine (NSN), Centre National de la Recherche Scientifique (CNRS), Institut Pasteur [Paris], Neurotransmission et sécrétion neuroendocrine ( NSN ), and Centre National de la Recherche Scientifique ( CNRS )

Subjects
Nervous system, rho GTP-Binding Proteins, actin cytoskeleton, Health, Toxicology and Mutagenesis, [SDV]Life Sciences [q-bio], [ SDV.TOX ] Life Sciences [q-bio]/Toxicology, MESH: Neurons, MESH : Actins, Apoptosis, Review, Toxicology, Synaptic Transmission, 0302 clinical medicine, Intestinal mucosa, MESH : Glutamic Acid, Neurotoxin, MESH: Animals, Intestinal Mucosa, toxin, Neurons, 0303 health sciences, MESH : Synaptic Transmission, MESH: Exocytosis, nervous system, MESH: Glutamic Acid, 3. Good health, Cell biology, medicine.anatomical_structure, MESH: Calcium, [SDV.TOX]Life Sciences [q-bio]/Toxicology, MESH : Monomeric GTP-Binding Proteins, Enterochromaffin cell, MESH: Intestinal Mucosa, SNARE Proteins, neurotransmitter, MESH: SNARE Proteins, Cell type, Bacterial Toxins, Neurotoxins, Glutamic Acid, MESH: Monomeric GTP-Binding Proteins, Biology, Blood–brain barrier, MESH: Actins, MESH : Neurotoxins, MESH : Intestinal Mucosa, MESH : Neurons, MESH : Exocytosis, Exocytosis, small gtpases, 03 medical and health sciences, MESH : rho GTP-Binding Proteins, medicine, MESH: Synaptic Transmission, Animals, Humans, MESH : Calcium, 030304 developmental biology, Monomeric GTP-Binding Proteins, MESH: Neurotoxins, MESH: Humans, [ SDV ] Life Sciences [q-bio], MESH: Apoptosis, MESH : Humans, neurotoxin, Actin cytoskeleton, MESH : SNARE Proteins, MESH: rho GTP-Binding Proteins, Actins, MESH: Bacterial Toxins, MESH : Bacterial Toxins, Enteric nervous system, Calcium, MESH : Animals, enterotoxin, 030217 neurology & neurosurgery, MESH : Apoptosis
Abstract

International audience; Toxins are potent molecules used by various bacteria to interact with a host organism. Some of them specifically act on neuronal cells (clostridial neurotoxins) leading to characteristics neurological affections. But many other toxins are multifunctional and recognize a wider range of cell types including neuronal cells. Various enterotoxins interact with the enteric nervous system, for example by stimulating afferent neurons or inducing neurotransmitter release from enterochromaffin cells which result either in vomiting, in amplification of the diarrhea, or in intestinal inflammation process. Other toxins can pass the blood brain barrier and directly act on specific neurons.

Published
2010
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10. Differences of Ca2+ handling properties in identified central olfactory neurons of the antennal lobe

Author

Andreas Husch, Peter Kloppenburg, Christophe Pouzat, Andreas Pippow, Department of Mouse Genetics and Metabolism, Laboratoire de physiologie cérébrale (LPC - UMR 8118), Université Paris Diderot - Paris 7 (UPD7)-Université Paris Descartes - Paris 5 (UPD5)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Centre National de la Recherche Scientifique (CNRS), Centre National de la Recherche Scientifique (CNRS)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Université Paris Diderot - Paris 7 (UPD7)-Université Paris Descartes - Paris 5 (UPD5), and Université Paris Diderot - Paris 7 (UPD7) - 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
Olfactory system, Patch-Clamp Techniques, Physiology, Action Potentials, Sodium Channels, chemistry.chemical_compound, 0302 clinical medicine, MESH: Smell, Periplaneta, MESH: Animals, MESH : Patch-Clamp Techniques, MESH: Action Potentials, Cells, Cultured, [INFO.INFO-BI] Computer Science [cs]/Bioinformatics [q-bio.QM], 0303 health sciences, [SDV.BIBS] Life Sciences [q-bio]/Quantitative Methods [q-bio.QM], biology, Depolarization, Anatomy, MESH: Interneurons, [SDV.BIBS]Life Sciences [q-bio]/Quantitative Methods [q-bio.QM], Smell, medicine.anatomical_structure, MESH: Calcium, MESH: Cells, Cultured, Fura-2, Calcium buffering, Olfaction, MESH : Interneurons, MESH: Sodium Channels, 03 medical and health sciences, MESH : Action Potentials, Interneurons, MESH : Cells, Cultured, MESH : Sodium Channels, MESH: Patch-Clamp Techniques, medicine, Animals, MESH : Calcium, Molecular Biology, 030304 developmental biology, Ion Transport, Cell Biology, biology.organism_classification, MESH: Ion Transport, Electrophysiology, chemistry, MESH : Periplaneta, Biophysics, MESH: Periplaneta, MESH : Smell, Antennal lobe, Calcium, MESH : Animals, [INFO.INFO-BI]Computer Science [cs]/Bioinformatics [q-bio.QM], MESH : Ion Transport, 030217 neurology & neurosurgery
Abstract

International audience; Information processing in neurons depends on highly localized Ca2+ signals. The spatial and temporal dynamics of these signals are determined by a variety of cellular parameters including the calcium influx, calcium buffering and calcium extrusion. Our long-term goal is to better understand how intracellular Ca2+ dynamics are controlled and contribute to information processing in defined interneurons of the insect olfactory system. The latter has served as an excellent model to study general mechanisms of olfaction. Using patch-clamp recordings and fast optical imaging in combination with the 'added buffer approach', we analyzed the Ca2+ handling properties of different identified neuron types in Periplaneta americana's olfactory system. Our focus was on two types of local interneurons (LNs) with significant differences in intrinsic electrophysiological properties: (1) spiking LNs that generate 'normal' Na+ driven action potentials and (2) non-spiking LNs that do not express voltage-activated Na+ channels. We found that the distinct electrophysiological properties from different types of central olfactory interneurons are strongly correlated with their cell specific calcium handling properties: non-spiking LNs, in which Ca2+ is the only cation that enters the cell to contribute to membrane depolarization, had the highest endogenous Ca2+ binding ratio and Ca2+ extrusion rate.

Published
2009
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11. Isolation and culture of term human cytotrophoblast cells and in vitro methods for studying human cytotrophoblast cells' calcium uptake

Author

Le Bellego , Frédérique, Vaillancourt , Cathy, Lafond , Julie, Institut Armand Frappier (INRS-IAF), Réseau International des Instituts Pasteur (RIIP)-Institut National de la Recherche Scientifique [Québec] (INRS), BioMed Research Centre [Montréal], Institut National de la Recherche Scientifique [Québec] (INRS)-Université du Québec à Trois-Rivières (UQTR)-Université du Québec à Montréal = University of Québec in Montréal (UQAM), Institut Armand Frappier ( INRS-IAF ), Institut National de la Recherche Scientifique [Québec] ( INRS ) -Réseau International des Instituts Pasteur ( RIIP ) -Institut Armand Frappier, BioMed Research Center, Department of Biological Sciences, and Université du Québec à Montréal ( UQAM )

Subjects
MESH: Cell Differentiation, MESH: Trophoblasts, Placenta, [ SDV.TOX ] Life Sciences [q-bio]/Toxicology, Fluorescent Antibody Technique, Enzyme-Linked Immunosorbent Assay, In Vitro Techniques, MESH : Cells, Cultured, Humans, MESH : Female, MESH : Calcium, MESH: Fluorescent Antibody Technique, Cells, Cultured, reproductive and urinary physiology, MESH: Humans, MESH : Humans, MESH: Enzyme-Linked Immunosorbent Assay, Cell Differentiation, MESH: Placenta, MESH : Enzyme-Linked Immunosorbent Assay, Trophoblasts, MESH : Fluorescent Antibody Technique, MESH : Placenta, MESH: Calcium, [SDV.TOX]Life Sciences [q-bio]/Toxicology, embryonic structures, Calcium, Female, MESH : Trophoblasts, MESH : Cell Differentiation, MESH: Female, MESH: Cells, Cultured
Abstract

National audience; Human primary cytotrophoblast cell culture is a very useful model to study the endocrine and immunological functions of syncytiotrophoblasts, as well as the ion exchange between the mother and her fetus, like calcium. In this chapter, we expose the procedure to (1) isolate and purify the cytotrophoblast cells from human term placenta and (2) study syncytiotrophoblast calcium uptake. First, the methodology is based on the enzymatic dissociation of villous placental tissue, followed by Percoll gradient separation. Purity is assessed by flow cytometry using staining against cytokeratin-7, protein specific for trophoblast cells. Cell proliferation is evaluated by a Thiazolyl Blue Tetrazolium Bromide (MTT) assay, hormonal secretion is measured by enzyme-linked immunosorbent assay (ELISA), and fusion is estimated by immunofluorescence using staining against desmosomal proteins. Second, we describe the calcium uptake experiment using the cytotrophoblast cells in culture.

Published
2009
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12. Wide-field and two-photon imaging of brain activity with voltage- and calcium-sensitive dyes

Author

Lawrence B. Cohen, Arthur Konnerth, Olga Garaschuk, Bradley J. Baker, Marco Canepari, Ryota Homma, Dejan Zecevic, Maja Djurisic, Lei Jin, Chun X. Bleau, Department of Cellular and Molecular Physiology, Yale School of Medicine, Institute of Neuroscience, Technical University of Munich ( TUM ), RedShirtImaging, LLC, DC05259/DC/NIDCD NIH HHS/United States NS42739/NS/NINDS NIH HHS/United States, Fahmeed Hyder, Yale University School of Medicine, and Technische Universität Munchen - Université Technique de Munich [Munich, Allemagne] (TUM)

Subjects
MESH: Photons, Spike train, Population, Biology, Noise (electronics), MESH: Brain, 03 medical and health sciences, 0302 clinical medicine, MESH : Photons, Two-photon excitation microscopy, MESH : Coloring Agents, [SDV.MHEP.PHY]Life Sciences [q-bio]/Human health and pathology/Tissues and Organs [q-bio.TO], medicine, MESH: Animals, education, MESH : Calcium, 030304 developmental biology, 0303 health sciences, education.field_of_study, MESH: Humans, Olfactory receptor, MESH : Evoked Potentials, [ SDV.MHEP.PHY ] Life Sciences [q-bio]/Human health and pathology/Tissues and Organs [q-bio.TO], MESH : Humans, Shot noise, Animals Brain/*physiology Calcium/*chemistry Coloring Agents/*administration & dosage Evoked Potentials Humans *Photons, Olfactory bulb, MESH: Evoked Potentials, Light intensity, medicine.anatomical_structure, MESH : Brain, MESH: Calcium, Biophysics, MESH : Animals, MESH: Coloring Agents, 030217 neurology & neurosurgery
Abstract

International audience; This chapter presents three examples of imaging brain activity with voltage- or calcium-sensitive dyes. Because experimental measurements are limited by low sensitivity, the chapter then discusses the methodological aspects that are critical for optimal signal-to-noise ratio. Two of the examples use wide-field (1-photon) imaging and the third uses two-photon scanning microscopy. These methods have relatively high temporal resolution ranging from 10 to 10,000 Hz. The three examples are the following: (1) Internally injected voltage-sensitive dye can be used to monitor membrane potential in the dendrites of invertebrate and vertebrate neurons in in vitro preparations. These experiments are directed at understanding how individual neurons convert the complex input synaptic activity into the output spike train. (2) Recently developed methods for staining many individual cells in the mammalian brain with calcium-sensitive dyes together with two-photon microscopy made it possible to follow the spike activity of many neurons simultaneously while in vivo preparations are responding to stimulation. (3) Calcium-sensitive dyes that are internalized into olfactory receptor neurons in the nose will, after several days, be transported to the nerve terminals of these cells in the olfactory bulb glomeruli. There, the population signals can be used as a measure of the input from the nose to the bulb. Three kinds of noise in measuring light intensity are discussed: (1) Shot noise from the random emission of photons from the preparation. (2) Extraneous (technical) noise from external sources. (3) Noise that occurs in the absence of light, the dark noise. In addition, we briefly discuss the light sources, the optics, and the detectors and cameras. The commonly used organic voltage and ion sensitive dyes stain all of the cell types in the preparation indiscriminately. A major effort is underway to find methods for staining individual cell types in the brain selectively. Most of these efforts center around fluorescent protein activity sensors because transgenic methods can be used to express them in individual cell types.

Published
2009
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13. Wide-field and two-photon imaging of brain activity with voltage- and calcium-sensitive dyes. : Imaging brain activity

Author

Homma, Ryota, Baker, Bradley, Jin, Lei, Garaschuk, Olga, Konnerth, Arthur, Cohen, Lawrence, Bleau, Chun, Canepari, Marco, Djurisic, Maja, Zecevic, Dejan, Department of Cellular and Molecular Physiology, Yale School of Medicine, Institute of Neuroscience, Technical University of Munich ( TUM ), RedShirtImaging, LLC, DC05259/DC/NIDCD NIH HHS/United States NS42739/NS/NINDS NIH HHS/United States, Fahmeed Hyder, Yale University School of Medicine, and Technische Universität Munchen - Université Technique de Munich [Munich, Allemagne] (TUM)

Subjects
MESH: Photons, MESH: Humans, MESH : Evoked Potentials, [ SDV.MHEP.PHY ] Life Sciences [q-bio]/Human health and pathology/Tissues and Organs [q-bio.TO], MESH : Humans, Animals Brain/*physiology Calcium/*chemistry Coloring Agents/*administration & dosage Evoked Potentials Humans *Photons, MESH: Brain, MESH: Evoked Potentials, MESH : Photons, MESH : Brain, MESH : Coloring Agents, MESH: Calcium, [SDV.MHEP.PHY]Life Sciences [q-bio]/Human health and pathology/Tissues and Organs [q-bio.TO], MESH: Animals, MESH : Animals, MESH : Calcium, MESH: Coloring Agents
Abstract

International audience; This chapter presents three examples of imaging brain activity with voltage- or calcium-sensitive dyes. Because experimental measurements are limited by low sensitivity, the chapter then discusses the methodological aspects that are critical for optimal signal-to-noise ratio. Two of the examples use wide-field (1-photon) imaging and the third uses two-photon scanning microscopy. These methods have relatively high temporal resolution ranging from 10 to 10,000 Hz. The three examples are the following: (1) Internally injected voltage-sensitive dye can be used to monitor membrane potential in the dendrites of invertebrate and vertebrate neurons in in vitro preparations. These experiments are directed at understanding how individual neurons convert the complex input synaptic activity into the output spike train. (2) Recently developed methods for staining many individual cells in the mammalian brain with calcium-sensitive dyes together with two-photon microscopy made it possible to follow the spike activity of many neurons simultaneously while in vivo preparations are responding to stimulation. (3) Calcium-sensitive dyes that are internalized into olfactory receptor neurons in the nose will, after several days, be transported to the nerve terminals of these cells in the olfactory bulb glomeruli. There, the population signals can be used as a measure of the input from the nose to the bulb. Three kinds of noise in measuring light intensity are discussed: (1) Shot noise from the random emission of photons from the preparation. (2) Extraneous (technical) noise from external sources. (3) Noise that occurs in the absence of light, the dark noise. In addition, we briefly discuss the light sources, the optics, and the detectors and cameras. The commonly used organic voltage and ion sensitive dyes stain all of the cell types in the preparation indiscriminately. A major effort is underway to find methods for staining individual cell types in the brain selectively. Most of these efforts center around fluorescent protein activity sensors because transgenic methods can be used to express them in individual cell types.

Published
2009
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14. Asp218 participates with Asp213 to bind a Ca2+ atom into the S1 subsite of aminopeptidase A: a key element for substrate specificity

Author

Bernard Maigret, Xavier Iturrioz, Mayumi Okada, Nicolas Inguimbert, Catherine Llorens-Cortes, Bernard P. Roques, Cédric Claperon, Raphael Rozenfeld, Neuropeptides centraux et régulations hydrique et cardiovasculaire, Université Pierre et Marie Curie - Paris 6 (UPMC)-Centre interdisciplinaire de recherche en biologie (CIRB), Labex MemoLife, École normale supérieure - Paris (ENS-PSL), Université Paris sciences et lettres (PSL)-Université Paris sciences et lettres (PSL)-Collège de France (CdF (institution))-Ecole Superieure de Physique et de Chimie Industrielles de la Ville de Paris (ESPCI Paris), Université Paris sciences et lettres (PSL)-École normale supérieure - Paris (ENS-PSL), Université Paris sciences et lettres (PSL)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Centre National de la Recherche Scientifique (CNRS)-Labex MemoLife, Université Paris sciences et lettres (PSL)-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), Unité de Pharmacologie Chimique et Génétique (UPCG - UMR_S 640/UMR 8151), Ecole Nationale Supérieure de Chimie de Paris - Chimie ParisTech-PSL (ENSCP), Université Paris sciences et lettres (PSL)-Université Paris sciences et lettres (PSL)-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)-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)-Institut de Chimie du CNRS (INC)-Centre National de la Recherche Scientifique (CNRS), Knowledge representation, reasonning (ORPAILLEUR), INRIA Lorraine, Institut National de Recherche en Informatique et en Automatique (Inria)-Institut National de Recherche en Informatique et en Automatique (Inria)-Laboratoire Lorrain de Recherche en Informatique et ses Applications (LORIA), Institut National de Recherche en Informatique et en Automatique (Inria)-Université Henri Poincaré - Nancy 1 (UHP)-Université Nancy 2-Institut National Polytechnique de Lorraine (INPL)-Centre National de la Recherche Scientifique (CNRS)-Université Henri Poincaré - Nancy 1 (UHP)-Université Nancy 2-Institut National Polytechnique de Lorraine (INPL)-Centre National de la Recherche Scientifique (CNRS), Université Pierre et Marie Curie - Paris 6 (UPMC)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Centre interdisciplinaire de recherche en biologie (CIRB), École normale supérieure - Paris (ENS Paris), Université Paris sciences et lettres (PSL)-École normale supérieure - Paris (ENS Paris), Université Paris sciences et lettres (PSL)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Centre National de la Recherche Scientifique (CNRS), Université Paris sciences et lettres (PSL)-Centre National de la Recherche Scientifique (CNRS)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Labex MemoLife, Université Paris sciences et lettres (PSL)-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)-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)-Institut de Chimie du CNRS (INC), Centre National de la Recherche Scientifique (CNRS)-Institut National Polytechnique de Lorraine (INPL)-Université Nancy 2-Université Henri Poincaré - Nancy 1 (UHP)-Institut National de Recherche en Informatique et en Automatique (Inria)-Centre National de la Recherche Scientifique (CNRS)-Institut National Polytechnique de Lorraine (INPL)-Université Nancy 2-Université Henri Poincaré - Nancy 1 (UHP), Université Pierre et Marie Curie - Paris 6 ( UPMC ) -Collège de France ( CdF ) -Institut National de la Santé et de la Recherche Médicale ( INSERM ), Unité de Pharmacologie Chimique et Génétique ( UPCG - UMR_S 640/UMR 8151 ), 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 ) -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 ), Knowledge representation, reasonning ( ORPAILLEUR ), Institut National de Recherche en Informatique et en Automatique ( Inria ) -Institut National de Recherche en Informatique et en Automatique ( Inria ) -Laboratoire Lorrain de Recherche en Informatique et ses Applications ( LORIA ), Institut National de Recherche en Informatique et en Automatique ( Inria ) -Université Henri Poincaré - Nancy 1 ( UHP ) -Université Nancy 2-Institut National Polytechnique de Lorraine ( INPL ) -Centre National de la Recherche Scientifique ( CNRS ) -Université Henri Poincaré - Nancy 1 ( UHP ) -Université Nancy 2-Institut National Polytechnique de Lorraine ( INPL ) -Centre National de la Recherche Scientifique ( CNRS ), Laboratoire Lorrain de Recherche en Informatique et ses Applications (LORIA), Institut National de Recherche en Informatique et en Automatique (Inria)-Université de Lorraine (UL)-Centre National de la Recherche Scientifique (CNRS), Iturrioz, Xavier, and Llorens Cortes, Catherine

Subjects
Models, Molecular, MESH : Glutamyl Aminopeptidase, MESH : Glutamates, MESH: Amino Acids, [SDV]Life Sciences [q-bio], Angiotensin III, monozinc aminopeptidase, MESH : Aspartic Acid, MESH: Cricetinae, MESH: Amino Acid Sequence, Biochemistry, MESH: Aspartic Acid, Substrate Specificity, MESH : Sulfhydryl Compounds, Mice, chemistry.chemical_compound, Glutamates, MESH: Cricetulus, Cricetinae, [CHIM] Chemical Sciences, MESH : CHO Cells, MESH: Animals, Amino Acids, Enzyme Inhibitors, Site-directed mutagenesis, inhibitor design, [SDV.MHEP.EM] Life Sciences [q-bio]/Human health and pathology/Endocrinology and metabolism, [PHYS]Physics [physics], MESH: Glutamyl Aminopeptidase, 0303 health sciences, MESH : Substrate Specificity, MESH: Kinetics, Hydrogen bond, MESH : Amino Acid Sequence, 030302 biochemistry & molecular biology, MESH : Cricetinae, Life Sciences, [ SDV.MHEP.EM ] Life Sciences [q-bio]/Human health and pathology/Endocrinology and metabolism, [SDV.MHEP.EM]Life Sciences [q-bio]/Human health and pathology/Endocrinology and metabolism, Phosphonate, MESH : Mutagenesis, Site-Directed, molecular modelling, [SDV] Life Sciences [q-bio], MESH: Mutagenesis, Site-Directed, Ectodomain, MESH: Enzyme Inhibitors, MESH: Calcium, Pharmacophore, MESH : Kinetics, site-directed mutagenesis, MESH : Phosphonic Acids, MESH: Models, Molecular, calcium-binding site, hypertension, Stereochemistry, MESH : Models, Molecular, education, MESH : Cricetulus, Organophosphonates, MESH: Phosphonic Acids, CHO Cells, Glutamyl Aminopeptidase, 03 medical and health sciences, Cricetulus, MESH: Glutamates, site directed mutagenesis, MESH: CHO Cells, MESH : Mice, Animals, [CHIM]Chemical Sciences, Amino Acid Sequence, Sulfhydryl Compounds, Carboxylate, calcium binding site, MESH : Calcium, MESH: Sulfhydryl Compounds, Molecular Biology, MESH: Mice, 030304 developmental biology, Aspartic Acid, MESH : Enzyme Inhibitors, Binding Sites, molecular modeling, Rational design, Cell Biology, Kinetics, chemistry, MESH: Binding Sites, Mutagenesis, Site-Directed, Calcium, MESH : Amino Acids, MESH: Substrate Specificity, MESH : Animals, MESH : Binding Sites
Abstract

International audience; APA (aminopeptidase A; EC 3.4.11.7) is a membrane-bound zinc metallopeptidase, also activated by Ca(2+), involved in the formation of brain angiotensin III, which exerts a tonic stimulatory action on the central control of blood pressure in hypertensive animals. In the present study, in the three-dimensional model of the ectodomain of mouse APA, we docked the specific APA inhibitor glutamate phosphonate, in the presence of Ca(2+). The model showed the presence of one Ca(2+) atom in an hydrophilic pocket corresponding to the S1 subsite in which the lateral chain of the inhibitor is pointing. In this pocket, the Ca(2+) atom was hexaco-ordinated with the acidic side chains of Asp(213) and Asp(218), the carbonyl group of Glu(215) and three water molecules, one of them being engaged in a hydrogen bond with the negatively charged carboxylate side chain of the inhibitor. Mutagenic replacement of Asp(213) and Asp(218) with a conservative residue maintained the ability of mutated APAs to be activated by Ca(2+). However, the replacement by a non-conservative residue abolished this property, demonstrating the crucial role of these residues in Ca(2+) binding. We also showed the involvement of these residues in the strict specificity of APA in the presence of Ca(2+) for N-terminal acidic residues from substrates or inhibitors, since mutagenic replacement of Asp(213) and Asp(218) induced a decrease of the inhibitory potencies of inhibitors homologous with acidic residues. Finally, this led to the rational design of a new potent APA inhibitor, NI926 (K(i)=70 nM), which allowed us to precisely localize Asp(213) at the entrance and Asp(218) at the bottom of the S1 subsite. Taken together, these data provide new insight into the organization and functional role of the APA S1 subsite and will allow the design of pharmacophore of the inhibitor, helpful for the development of a new generation of APA inhibitors as central-acting antihypertensive agents.

Published
2008
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15. Differential expression of parathyroid hormone-related protein in adrenocortical tumors: autocrine/paracrine effects on the growth and signaling pathways in H295R cells

Author

Fernande René-Corail, Karine Perlemoine, Marthe Rizk-Rabin, Xavier Bertagna, Frédérique Tissier, Jérôme Bertherat, Michèle Lieberherr, Guillaume Assié, Z. Bouizar, Hinda Hamzaoui, Institut Cochin (UMR_S567 / UMR 8104), Institut National de la Santé et de la Recherche Médicale (INSERM)-Centre National de la Recherche Scientifique (CNRS)-Université Paris Descartes - Paris 5 (UPD5), Pôle Endocrinologie-Diabétologie Adultes-Enfants, Assistance publique - Hôpitaux de Paris (AP-HP) (APHP)-CHU Cochin [AP-HP]-Hôpital Saint-Vincent de Paul, Nutrition et Régulation Lipidique des Fonctions Cérébrales, Institut National de la Recherche Agronomique (INRA), Service de biochimie-hormonologie [Béclère], Université Paris-Sud - Paris 11 (UP11)-Assistance publique - Hôpitaux de Paris (AP-HP) (APHP)-Hôpital Antoine Béclère, This study was supported in part by Plan Hospitalier de Recherche Clinique (PHRC) grant (AOM 06 179 to the Comete Network coordinated by Prof. P.F. Plouin) and by grants from INSERM and the Ministère Délégué à la Recherche et des Nouvelles Technologies. H. Hamzaoui was the recipient of fellowships from the Cancer & Solidarity Foundation, the Association pour la Recherche contre le Cancer (ARC) and the Fondation pour la Recherche Médicale (FRM)., Université Paris Descartes - Paris 5 (UPD5) - Institut National de la Santé et de la Recherche Médicale (INSERM) - Centre National de la Recherche Scientifique (CNRS), Assistance publique - Hôpitaux de Paris (AP-HP) - CHU Cochin [AP-HP] - Hôpital Saint-Vincent de Paul, Université Paris-Sud - Paris 11 (UP11) - Assistance publique - Hôpitaux de Paris (AP-HP) - Hôpital Antoine Béclère, Université Paris Descartes - Paris 5 (UPD5)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Centre National de la Recherche Scientifique (CNRS), Assistance publique - Hôpitaux de Paris (AP-HP) (AP-HP)-Hôpital Cochin [AP-HP], Assistance publique - Hôpitaux de Paris (AP-HP) (AP-HP)-Hôpital Saint-Vincent de Paul, AP-HP - Hôpital Antoine Béclère [Clamart], Assistance publique - Hôpitaux de Paris (AP-HP) (AP-HP)-Assistance publique - Hôpitaux de Paris (AP-HP) (AP-HP)-Université Paris-Sud - Paris 11 (UP11), Bouizar, Zhor, Université Paris-Sud - Paris 11 (UP11)-Assistance publique - Hôpitaux de Paris (AP-HP) (AP-HP)-AP-HP - Hôpital Antoine Béclère [Clamart], and Assistance publique - Hôpitaux de Paris (AP-HP) (AP-HP)

Subjects
Male, MESH: Signal Transduction, MESH : RNA, Messenger, Epidemiology, MESH : Aged, MESH : Adrenal Cortex Neoplasms, Apoptosis, MESH: Cell Cycle, MESH : Blotting, Western, MESH: Adrenal Cortex Neoplasms, Metastasis, 0302 clinical medicine, MESH: Reverse Transcriptase Polymerase Chain Reaction, MESH : Female, Receptor, MESH: Aged, 0303 health sciences, MESH : Prognosis, MESH: Middle Aged, Reverse Transcriptase Polymerase Chain Reaction, Paracrine cAMP, Cell Cycle, MESH : Reverse Transcriptase Polymerase Chain Reaction, Middle Aged, MESH : Adult, Prognosis, musculoskeletal system, 3. Good health, Blot, MESH : Cyclic AMP-Dependent Protein Kinases, qPCR, Oncology, 030220 oncology & carcinogenesis, MESH: Calcium, Female, Signal transduction, PTHrP Receptor, MESH : Parathyroid Hormone-Related Protein, hormones, hormone substitutes, and hormone antagonists, Signal Transduction, Adult, musculoskeletal diseases, medicine.medical_specialty, MESH: Cell Line, Tumor, Adolescent, MESH : Male, PTHrP, Blotting, Western, Ca++ signaling Pathways, [SDV.CAN]Life Sciences [q-bio]/Cancer, MESH: Cyclic AMP-Dependent Protein Kinases, Biology, MESH: Parathyroid Hormone-Related Protein, MESH: Prognosis, Adrenocortical Tumors, 03 medical and health sciences, Paracrine signalling, MESH : Receptor, Parathyroid Hormone, Type 1, [SDV.CAN] Life Sciences [q-bio]/Cancer, Internal medicine, Cell Line, Tumor, MESH : Adolescent, MESH : Cell Cycle, medicine, Didderential Expression, Humans, MESH: Blotting, Western, MESH : Middle Aged, RNA, Messenger, Autocrine, Autocrine signalling, Protein kinase A, MESH : Calcium, Protein kinase C, 030304 developmental biology, Aged, Receptor, Parathyroid Hormone, Type 1, MESH: RNA, Messenger, MESH : Signal Transduction, MESH: Adolescent, MESH: Humans, Parathyroid hormone-related protein, MESH : Cell Line, Tumor, MESH: Apoptosis, MESH : Humans, Parathyroid Hormone-Related Protein, Antagonist, MESH: Adult, Cyclic AMP-Dependent Protein Kinases, Adrenal Cortex Neoplasms, MESH: Male, Endocrinology, MESH: Receptor, Parathyroid Hormone, Type 1, Cancer research, Calcium, MESH: Female, MESH : Apoptosis
Abstract

Adrenocortical tumors (ACT) are rare and heterogeneous, but their pathogenesis is unclear. The oncoprotein parathyroid hormone–related protein (PTHrP), found in many common tumors, can regulate their growth in an autocrine/paracrine fashion through the PTH-R1 receptor. Little is known about the role of PTHrP in ACT. We monitored the synthesis of PTHrP and PTH-R1 in a series of 25 ACT: 12 adrenocortical carcinomas (ACC) and 13 adrenocortical adenomas (ACA), and investigated the effects of PTHrP(1-34) on H295R cells derived from an ACC. PTH-R1 mRNA and proteins were detected by real-time PCR and Western blotting in all the ACT samples and in H295R cells. Their concentrations did not differ significantly from one ACT to another. PTHrP mRNA was assayed by quantitative real-time PCR. It was detected in 90% of ACC, and in 10% of ACA. There was a positive correlation with the prognostic factors, McFarlane stage and Weiss score. Tissue-specific PTHrP protein processing was shown by Western blotting. Immunohistochemical staining revealed numerous, dense foci of PTHrP-containing cells in ACC, but few positive cells in ACA or normal tissue. PTHrP stimulated the growth of H295R cells, whereas a specific anti-PTHrP antibody and a PTHrP-R1 antagonist both enhanced their apoptosis. PTHrP activated both adenylate cyclase/protein kinase A and the intracellular calcium/protein kinase C pathways via PTHrP-R1. The active synthesis of PTHrP is linked to poor prognosis in ACC, in which it may act as an autocrine/paracrine factor in tumor growth and malignancy. (Cancer Epidemiol Biomarkers Prev 2008;17(9):2275–85)

Published
2008
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16. A novel druglike spleen tyrosine kinase binder prevents anaphylactic shock when administered orally

Author

Bruno O. Villoutreix, Thomas Roumier, Elsa Mazuc, Jean-Paul Leonetti, Odile Malbec, David Dombrowicz, Sébastien Fleury, Marc Daëron, Pierre Martineau, Piona Dariavach, Institut de recherche en cancérologie de Montpellier ( IRCM - U896 Inserm - UM1 ), Université Montpellier 1 ( UM1 ) -CRLCC Val d'Aurelle - Paul Lamarque-Institut National de la Santé et de la Recherche Médicale ( INSERM ) -Université de Montpellier ( UM ), Pharmacochimie Moléculaire et Cellulaire ( PMC - UMR_S 648 ), Centre National de la Recherche Scientifique ( CNRS ) -Institut National de la Santé et de la Recherche Médicale ( INSERM ) -Université Paris Descartes - Paris 5 ( UPD5 ), Allergologie Moléculaire et Cellulaire, Institut Pasteur [Paris]-Institut National de la Santé et de la Recherche Médicale ( INSERM ), Schistosomiase, paludisme et inflammation, Institut Pasteur de Lille, Réseau International des Instituts Pasteur ( RIIP ) -Réseau International des Instituts Pasteur ( RIIP ) -Institut National de la Santé et de la Recherche Médicale ( INSERM ) -Université de Lille, Droit et Santé, Institut de Recherche en Infectiologie de Montpellier ( IRIM ), Université de Montpellier ( UM ) -Centre National de la Recherche Scientifique ( CNRS ), This work was supported by Languedoc Roussillon Incubation, the INSERM Avenir award, CNRS and the University of Montpellier., Institut de recherche en cancérologie de Montpellier (IRCM - U896 Inserm - UM1), Université Montpellier 1 (UM1)-CRLCC Val d'Aurelle - Paul Lamarque-Institut National de la Santé et de la Recherche Médicale (INSERM)-Université de Montpellier (UM), Pharmacochimie Moléculaire et Cellulaire (PMC - UMR_S 648), 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 Pasteur [Paris] (IP)-Institut National de la Santé et de la Recherche Médicale (INSERM), Réseau International des Instituts Pasteur (RIIP)-Réseau International des Instituts Pasteur (RIIP)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Université de Lille, Droit et Santé, Institut de Recherche en Infectiologie de Montpellier (IRIM), Université de Montpellier (UM)-Centre National de la Recherche Scientifique (CNRS), 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 Pasteur [Paris]-Institut National de la Santé et de la Recherche Médicale (INSERM), Le Ster, Yves, CRLCC Val d'Aurelle - Paul Lamarque-Université de Montpellier (UM)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Université Montpellier 1 (UM1), and Centre National de la Recherche Scientifique (CNRS)-Université de Montpellier (UM)

Subjects
Pharmacology, Anaphylactic shock, Cell Degranulation, MESH: Antibodies, Monoclonal, Mice, 0302 clinical medicine, Immunoreceptor tyrosine-based activation motif, Medicine, [ SDV.IMM ] Life Sciences [q-bio]/Immunology, MESH: Animals, MESH : Receptors, IgE, 0303 health sciences, Intracellular Signaling Peptides and Proteins, Antibodies, Monoclonal, 3. Good health, MESH: Administration, Oral, MESH : Mast Cells, MESH: Passive Cutaneous Anaphylaxis, Tyrosine kinase, MESH: Enzyme Activation, [SDV.IMM] Life Sciences [q-bio]/Immunology, Immunology, Drug-like compound, MESH : Cell Degranulation, MESH: Receptors, IgE, MESH: Protein-Tyrosine Kinases, 03 medical and health sciences, MESH : Protein-Tyrosine Kinases, Anaphylaxis, MESH : Anaphylaxis, Degranulation, MESH : Mitogen-Activated Protein Kinases, Tyrosine kinase Syk, medicine.disease, Enzyme Activation, Thiazoles, [ SDV.SP ] Life Sciences [q-bio]/Pharmaceutical sciences, Allergy and Inflammation, MESH: Female, Type I hypersensitivity, MESH: Signal Transduction, Administration, Oral, Syk, Proto-Oncogene Proteins c-fyn, medicine.disease_cause, MESH : Thiazoles, Immunology and Allergy, MESH : Female, Mast Cells, Mice, Inbred BALB C, MESH : Proto-Oncogene Proteins c-fyn, Passive Cutaneous Anaphylaxis, MESH: Mast Cells, Protein-Tyrosine Kinases, [SDV.SP]Life Sciences [q-bio]/Pharmaceutical sciences, Mast cells/basophils, Mast cell, [SDV.SP] Life Sciences [q-bio]/Pharmaceutical sciences, src-Family Kinases, medicine.anatomical_structure, Biochemistry, MESH: Calcium, MESH : Antibodies, Monoclonal, MESH: Cell Degranulation, Allergic response, [SDV.IMM]Life Sciences [q-bio]/Immunology, Female, Mitogen-Activated Protein Kinases, Signal Transduction, MESH: Mice, Inbred BALB C, MESH: Thiazoles, Linker for Activation of T cells, MESH : Intracellular Signaling Peptides and Proteins, MESH: Intracellular Signaling Peptides and Proteins, MESH : Mice, Animals, Syk Kinase, MESH : Calcium, MESH: Mice, MESH : Mice, Inbred BALB C, 030304 developmental biology, MESH : Signal Transduction, MESH : Administration, Oral, Receptors, IgE, business.industry, MESH: Proto-Oncogene Proteins c-fyn, MESH: Mitogen-Activated Protein Kinases, MESH: Anaphylaxis, MESH: src-Family Kinases, MESH : Passive Cutaneous Anaphylaxis, Calcium, MESH : src-Family Kinases, MESH : Animals, business, MESH : Enzyme Activation, 030215 immunology
Abstract

International audience; BACKGROUND: The spleen tyrosine kinase (Syk) is recognized as a potential pharmaceutical target for the treatment of type I hypersensitivity reactions including allergic rhinitis, urticaria, asthma, and anaphylaxis because of its critical position upstream of immunoreceptor signaling complexes that regulate inflammatory responses in leukocytes. OBJECTIVE: Our aim was to improve the selectivity of anti-Syk therapies by impeding the interaction of Syk with its cellular partners, instead of targeting its catalytic site. METHODS: We have previously studied the inhibitory effects of the anti-Syk intracellular antibody G4G11 on Fc epsilonRI-induced release of allergic mediators. A compound collection was screened by using an antibody displacement assay to identify functional mimics of G4G11 that act as potential inhibitors of the allergic response. The effects of the selected druglike compounds on mast cell activation were evaluated in vitro and in vivo. RESULTS: We discovered compound 13, a small molecule that inhibits Fc epsilonRI-induced mast cell degranulation in vitro and anaphylactic shock in vivo. Importantly, compound 13 was efficient when administered orally to mice. Structural analysis, docking, and site-directed mutagenesis allowed us to identify the binding cavity of this compound, located at the interface between the 2 Src homology 2 domains and the interdomain A of Syk. CONCLUSION: We have isolated a new class of druglike compounds that modulate the interaction of Syk with some of its macromolecular substrates implicated in the degranulation pathway in mast cells.

Published
2008
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17. Cardiomyocyte overexpression of neuronal nitric oxide synthase delays transition toward heart failure in response to pressure overload by preserving calcium cycling

Author

Estelle Robidel, Bernd Mayer, Christophe Heymes, Laurent Vinet, Emilie Vaudin, Ana María Gómez, María Fernández-Velasco, Xavier Loyer, Paul Milliez, Dominique Charue, Jean-Jacques Mercadier, Ajay M. Shah, Frederic Jaisser, Sylvain Richard, Isabelle Marty, Yannis Sainte-Marie, Jane-Lise Samuel, Jean-Pierre Benitah, Peter Vangheluwe, Wei Zhang, Centre de Recherche Cardiovasculaire de Lariboisiere, Université Paris Diderot - Paris 7 (UPD7)-Institut National de la Santé et de la Recherche Médicale (INSERM), Physiopathologie cardiovasculaire, Université Montpellier 1 (UM1)-IFR3, Université Montpellier 1 (UM1)-Institut National de la Santé et de la Recherche Médicale (INSERM), Laboratory of Ca-transport ATPases, The James Black Centre, Hémostase, bio-ingénierie et remodelage cardiovasculaires (LBPC), Université Paris 13 (UP13)-Université Paris Diderot - Paris 7 (UPD7)-Institut Galilée-Université Sorbonne Paris Cité (USPC)-Institut National de la Santé et de la Recherche Médicale (INSERM), Grenoble Institut des Neurosciences (GIN), Université Joseph Fourier - Grenoble 1 (UJF)-Institut National de la Santé et de la Recherche Médicale (INSERM), Department of Pharmacology and Toxicology, Karl-Franzens-Universität Graz, Gènes et pression artérielle (INSERM U772), Collège de France (CdF (institution))-Université Paris Descartes - Paris 5 (UPD5)-Institut National de la Santé et de la Recherche Médicale (INSERM), Cardiovascular Division, King‘s College London, Roux-Buisson, Nathalie, Université Paris Diderot - Paris 7 (UPD7)-Université Paris 13 (UP13)-Université Sorbonne Paris Cité (USPC)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Institut Galilée, Université Paris Diderot - Paris 7 ( UPD7 ) -Institut National de la Santé et de la Recherche Médicale ( INSERM ), Université Montpellier 1 ( UM1 ) -IFR3-Institut National de la Santé et de la Recherche Médicale ( INSERM ), Hémostase, bio-ingénierie et remodelage cardiovasculaires ( LBPC ), Université Paris Diderot - Paris 7 ( UPD7 ) -Université Paris 13 ( UP13 ) -Université Sorbonne Paris Cité ( USPC ) -Institut National de la Santé et de la Recherche Médicale ( INSERM ) -Institut Galilée, Grenoble Institut des Neurosciences ( GIN ), Université Joseph Fourier - Grenoble 1 ( UJF ) -CHU Grenoble-Institut National de la Santé et de la Recherche Médicale ( INSERM ), Gènes et pression artérielle ( Inserm U772 ), and Collège de France ( CdF ) -Université Paris Descartes - Paris 5 ( UPD5 ) -Institut National de la Santé et de la Recherche Médicale ( INSERM )

Subjects
MESH: Signal Transduction, MESH: Myocardial Contraction, heart failure, MESH: Myocytes, Cardiac, Blood Pressure, MESH: Nitric Oxide Synthase Type I, Cell Separation, Nitric Oxide Synthase Type I, 030204 cardiovascular system & hematology, Ventricular Function, Left, MESH: Ventricular Function, Left, Muscle hypertrophy, Mice, 0302 clinical medicine, MESH: Reverse Transcriptase Polymerase Chain Reaction, MESH : Nitric Oxide Synthase Type I, Myocyte, Myocytes, Cardiac, MESH: Animals, MESH : Myocardial Contraction, Aorta, remodeling, 0303 health sciences, biology, Reverse Transcriptase Polymerase Chain Reaction, nitric oxide synthase, MESH : Reverse Transcriptase Polymerase Chain Reaction, MESH: Aorta, MESH: Blood Pressure, MESH : Mice, Transgenic, Phospholamban, Nitric oxide synthase, MESH: Calcium, Disease Progression, MESH: Disease Progression, Cardiology and Cardiovascular Medicine, hypertrophy, Signal Transduction, MESH : Ventricular Function, Left, medicine.medical_specialty, MESH: Enzyme Activation, MESH: Mice, Transgenic, MESH : Myocytes, Cardiac, Concentric hypertrophy, Mice, Transgenic, MESH: Cell Separation, Contractility, 03 medical and health sciences, Physiology (medical), Internal medicine, MESH : Mice, medicine, MESH : Blood Pressure, [SDV.BBM] Life Sciences [q-bio]/Biochemistry, Molecular Biology, Animals, Humans, [SDV.BBM]Life Sciences [q-bio]/Biochemistry, Molecular Biology, MESH : Calcium, [ SDV.BBM ] Life Sciences [q-bio]/Biochemistry, Molecular Biology, MESH: Mice, 030304 developmental biology, Pressure overload, MESH : Signal Transduction, calcium, MESH: Humans, business.industry, MESH : Humans, MESH : Aorta, MESH : Disease Progression, medicine.disease, MESH : Disease Models, Animal, Myocardial Contraction, Enzyme Activation, Disease Models, Animal, Endocrinology, Heart failure, MESH : Cell Separation, MESH: Heart Failure, biology.protein, MESH : Animals, MESH: Disease Models, Animal, business, MESH : Enzyme Activation, MESH : Heart Failure
Abstract

Background— Defects in cardiomyocyte Ca 2+ cycling are a signature feature of heart failure (HF) that occurs in response to sustained hemodynamic overload, and they largely account for contractile dysfunction. Neuronal nitric oxide synthase (NOS1) influences myocyte excitation-contraction coupling through modulation of Ca 2+ cycling, but the potential relevance of this in HF is unknown. Methods and Results— We generated a transgenic mouse with conditional, cardiomyocyte-specific NOS1 overexpression (double-transgenic [DT]) and studied cardiac remodeling, myocardial Ca 2+ handling, and contractility in DT and control mice subjected to transverse aortic constriction (TAC). After TAC, control mice developed eccentric hypertrophy with evolution toward HF as revealed by a significantly reduced fractional shortening. In contrast, DT mice developed a greater increase in wall thickness ( P P P 2+ transients, and sarcoplasmic reticulum Ca 2+ load ( P Conclusions— Cardiomyocyte NOS1 may be a useful target against cardiac deterioration during chronic pressure-overload–induced HF through modulation of calcium cycling.

Published
2008
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18. Abnormalities of mitochondrial functioning can partly explain the metabolic disorders encountered in sarcopenic gastrocnemius

Author

Catherine Vergely, Laurent Mosoni, Hervé Dubouchaud, Luc Demaison, Eric Fontaine, Jean-Michel Chardigny, A. Oudot, Caroline Martin, Xavier Leverve, C. Keriel, Luc Rochette, Unité de nutrition lipidique, Institut National de la Recherche Agronomique (INRA), Bioénergétique fondamentale et appliquée, Université Joseph Fourier - Grenoble 1 (UJF)-Institut National de la Santé et de la Recherche Médicale (INSERM), Métabolisme protéino-énergétique (UMPE), Institut National de la Recherche Agronomique (INRA)-Université d'Auvergne - Clermont-Ferrand I (UdA), Laboratoire de Physiopathologie et de Pharmacologie Cardiovasculaire Expérimentale (LPPCE), Université de Bourgogne (UB), Hamant, Sarah, Institut National de la Recherche Agronomique ( INRA ), Université Joseph Fourier - Grenoble 1 ( UJF ) -Institut National de la Santé et de la Recherche Médicale ( INSERM ), Métabolisme protéino-énergétique ( UMPE ), Institut National de la Recherche Agronomique ( INRA ) -Université d'Auvergne - Clermont-Ferrand I ( UdA ), Laboratoire de Physiopathologie et de Pharmacologie Cardiovasculaire Expérimentale ( LPPCE ), Université de Bourgogne ( UB ), and Université d'Auvergne - Clermont-Ferrand I (UdA)-Institut National de la Recherche Agronomique (INRA)

Subjects
Male, muscle atrophy, MESH : Cell Aging, [SDV]Life Sciences [q-bio], MESH : Reactive Oxygen Species, Mitochondrion, 0302 clinical medicine, Glycolysis, MESH: Animals, MESH : Muscle, Skeletal, MESH : Fatty Acids, Cellular Senescence, Phospholipids, MESH: Superoxide Dismutase, reactive oxygen species, MESH : Free Radicals, 0303 health sciences, MESH: Muscle, Skeletal, MESH : Rats, Fatty Acids, fatty acid profile of mitochondrial lipids, MESH: Reactive Oxygen Species, Pyruvate dehydrogenase complex, MESH: Fatty Acids, mitochondria, [SDV] Life Sciences [q-bio], Biochemistry, MESH: Cell Aging, MESH: Calcium, MESH : Mitochondria, Cell aging, Pyruvate decarboxylation, medicine.medical_specialty, Free Radicals, MESH: Rats, Cellular respiration, MESH: Mitochondria, MESH : Male, Cell Respiration, chemistry.chemical_element, Oxidative phosphorylation, Biology, Calcium, MESH : Rats, Wistar, MESH : Phospholipids, 03 medical and health sciences, MESH: Free Radicals, Internal medicine, medicine, Animals, MESH : Superoxide Dismutase, Rats, Wistar, Muscle, Skeletal, MESH : Calcium, 030304 developmental biology, MESH: Phospholipids, calcium, permeability transition pore, Superoxide Dismutase, aging, Cell Aging, Mitochondria, Muscle, Skeletal, Rats, Wistar, Reactive Oxygen Species, Cell Biology, MESH: Rats, Wistar, MESH: Male, Endocrinology, chemistry, MESH : Cell Respiration, MESH : Animals, MESH: Cell Respiration, 030217 neurology & neurosurgery
Abstract

International audience; Aging triggers several abnormalities in muscle glycolytic fibers including increased proteolysis, reactive oxygen species (ROS) production and apoptosis. Since the mitochondria are the main site of substrate oxidation, ROS production and programmed cell death, we tried to know whether the cellular disorders encountered in sarcopenia are due to abnormal mitochondrial functioning. Gastrocnemius mitochondria were extracted from adult (6 months) and aged (21 months) male Wistar rats. Respiration parameters, opening of the permeability transition pore and ROS production, with either glutamate (amino acid metabolism) or pyruvate (glucose metabolism) as a respiration substrate, were evaluated at different matrix calcium concentrations. Pyruvate dehydrogenase and respiratory complex activities as well as their contents measured by Western blotting analysis were determined. Furthermore, the fatty acid profile of mitochondrial phospholipids was also measured. At physiological calcium concentration, state III respiration rate was lowered by aging in pyruvate conditions (-22%), but not with glutamate. The reduction of pyruvate oxidation resulted from a calcium-dependent inactivation of the pyruvate dehydrogenase system and could provide for the well-known proteolysis encountered during sarcopenia. Matrix calcium loading and aging increased ROS production. They also reduced the oxidative phosphorylation. This was associated with lower calcium retention capacities, suggesting that sarcopenic fibers are more prone to programmed cell death. Aging was also associated with a reduced mitochondrial superoxide dismutase activity, which does not intervene in toxic ROS overproduction but could explain the lower calcium retention capacities. Despite a lower content, cytochrome c oxidase displayed an increased activity associated with an increased n-6/n-3 polyunsaturated fatty acid ratio of mitochondrial phospholipids. In conclusion, we propose that mitochondria obtained from aged muscle fibers display several functional abnormalities explaining the increased proteolysis, ROS overproduction and vulnerability to apoptosis exhibited by sarcopenic muscle. These changes appear to be related to modifications of the fatty acid profile of mitochondrial lipids.

Published
2007
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19. Cellular localization of apelin and its receptor in the anterior pituitary: evidence for a direct stimulatory action of apelin on ACTH release

Author

Marie-Thérèse Bluet-Pajot, Rodrigo Alvear-Perez, Philippe Zizzari, Catherine Llorens-Cortes, Annabelle Réaux-Le Goazigo, Jacques Epelbaum, Neuropeptides centraux et régulations hydrique et cardiovasculaire, Université Pierre et Marie Curie - Paris 6 (UPMC)-Centre interdisciplinaire de recherche en biologie (CIRB), Labex MemoLife, École normale supérieure - Paris (ENS-PSL), Université Paris sciences et lettres (PSL)-Université Paris sciences et lettres (PSL)-Collège de France (CdF (institution))-Ecole Superieure de Physique et de Chimie Industrielles de la Ville de Paris (ESPCI Paris), Université Paris sciences et lettres (PSL)-École normale supérieure - Paris (ENS-PSL), Université Paris sciences et lettres (PSL)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Centre National de la Recherche Scientifique (CNRS)-Labex MemoLife, Université Paris sciences et lettres (PSL)-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), Neurobiologie de la Croissance et de la Senescence, 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 Pierre et Marie Curie - Paris 6 (UPMC)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Centre interdisciplinaire de recherche en biologie (CIRB), École normale supérieure - Paris (ENS Paris), Université Paris sciences et lettres (PSL)-École normale supérieure - Paris (ENS Paris), Université Paris sciences et lettres (PSL)-Centre National de la Recherche Scientifique (CNRS)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Labex MemoLife, Université Paris sciences et lettres (PSL)-Centre National de la Recherche Scientifique (CNRS)-Institut National de la Santé et de la Recherche Médicale (INSERM), Université Pierre et Marie Curie - Paris 6 ( UPMC ) -Collège de France ( CdF ) -Institut National de la Santé et de la Recherche Médicale ( INSERM ), Université Paris Descartes - Paris 5 ( UPD5 ) -Institut National de la Santé et de la Recherche Médicale ( INSERM ) -Centre National de la Recherche Scientifique ( CNRS ), and Llorens Cortes, Catherine

Subjects
Male, Pituitary gland, Endocrinology, Diabetes and Metabolism, MESH: Amino Acid Sequence, MESH: Receptors, G-Protein-Coupled, MESH: Base Sequence, MESH : Tissue Distribution, Receptors, G-Protein-Coupled, Rats, Sprague-Dawley, 0302 clinical medicine, MESH: Cricetulus, MESH: Pituitary Gland, MESH: Ligands, MESH : CHO Cells, Tissue Distribution, MESH: Animals, MESH: Peptide Fragments, MESH : Ligands, 0303 health sciences, MESH : Amino Acid Sequence, MESH : Cyclic AMP, MESH: Pituitary Gland, Anterior, MESH : Cricetinae, MESH : Protein Binding, [ SDV.MHEP.EM ] Life Sciences [q-bio]/Human health and pathology/Endocrinology and metabolism, [SDV.MHEP.EM]Life Sciences [q-bio]/Human health and pathology/Endocrinology and metabolism, Apelin, MESH : Adrenocorticotropic Hormone, Intercellular Signaling Peptides and Proteins, MESH : Carrier Proteins, medicine.medical_specialty, endocrine system, MESH : Cell Membrane, MESH : Cricetulus, MESH: Carrier Proteins, Adrenocorticotropic hormone, 03 medical and health sciences, Adrenocorticotropic Hormone, Anterior pituitary, Pituitary Gland, Anterior, MESH: CHO Cells, Physiology (medical), MESH: Protein Binding, MESH: Tissue Distribution, MESH: Adrenocorticotropic Hormone, MESH: Humans, MESH: Molecular Sequence Data, MESH : Humans, Peptide Fragments, Endocrinology, Carrier Proteins, 030217 neurology & neurosurgery, MESH : Molecular Sequence Data, Physiology, MESH: Corticotrophs, MESH: Cricetinae, MESH : Models, Biological, MESH: Rats, Sprague-Dawley, Rats, Inbred WKY, MESH : Pituitary Gland, Receptor, Corticotrophs, Cellular localization, MESH: Cyclic AMP, [SDV.MHEP.EM] Life Sciences [q-bio]/Human health and pathology/Endocrinology and metabolism, Apelin Receptors, MESH : Rats, MESH : Forskolin, medicine.anatomical_structure, MESH: Pituitary Hormones, Anterior, MESH: Calcium, MESH : Receptors, G-Protein-Coupled, MESH : Transfection, MESH: Forskolin, hormones, hormone substitutes, and hormone antagonists, MESH: Rats, Inbred WKY, Endocrine gland, MESH: Rats, MESH : Male, MESH : Rats, Inbred WKY, Biology, Models, Biological, MESH : Pituitary Hormones, Anterior, MESH : Pituitary Gland, Anterior, Pituitary Hormones, Anterior, Internal medicine, medicine, Animals, MESH : Calcium, 030304 developmental biology, Apelin receptor, MESH: Transfection, MESH : Peptide Fragments, MESH: Models, Biological, MESH : Corticotrophs, MESH : Rats, Sprague-Dawley, MESH: Male, Rats, MESH : Peptide Hormones, MESH: Peptide Hormones, MESH : Base Sequence, MESH : Animals, MESH: Cell Membrane
Abstract

Apelin is a bioactive peptide recently identified as the endogenous ligand of the human orphan G protein-coupled receptor APJ. The presence of apelin-immunoreactive nerve fibers, together with the detection of apelin receptor mRNA in the parvocellular part of the paraventricular nucleus and the stimulatory action of apelin on corticotropin-releasing hormone release, indicate that apelin modulates adrenocorticotropin (ACTH) release via an indirect action on the hypothalamus. However, a direct action of apelin in the anterior pituitary cannot be excluded. Here, we provided evidence for the existence of an apelinergic system within the adult male rat pituitary gland. Double immunofluorescence staining indicated that apelin is highly coexpressed in the anterior pituitary, mainly in corticotrophs (96.5 ± 0.3%) and to a much lower extent in somatotropes (3.2 ± 0.2%). Using in situ hybridization combined with immunohistochemistry, a high expression of apelin receptor mRNA was also found in corticotrophs, suggesting a local interaction between apelin and ACTH. In an ex vivo perifusion system of anterior pituitaries, apelin 17 (K17F, 10−6M) significantly increased basal ACTH release by 41%, whereas apelin 10 (R10F, 10−6M), an inactive apelin fragment, was ineffective. In addition, K17F but not R10F induced a dose-dependent increase in K+-evoked ACTH release, with maximal increase being observed for a 10−6M concentration. Taken together, these data outline the potential role of apelin as an autocrine/paracrine-acting peptide on ACTH release and provide morphological and neuroendocrine basis for further studies that explore the physiological role of apelin in the regulation of anterior pituitary functions.

Published
2007
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20. [The calcium, an object of study, astonishing, no?]

Author

Moreau, Marc, Ranjeva, Raoul, Laboratoire de Physique Théorique des Liquides (LPTL), Université Pierre et Marie Curie - Paris 6 (UPMC)-Centre National de la Recherche Scientifique (CNRS), Laboratoire de Géographie Physique et Environnementale (GEOLAB), Université Blaise Pascal - Clermont-Ferrand 2 (UBP)-Université Clermont Auvergne [2017-2020] (UCA [2017-2020])-Université Clermont Auvergne (UCA)-Institut Sciences de l'Homme et de la Société (IR SHS UNILIM), Université de Limoges (UNILIM)-Université de Limoges (UNILIM)-Centre National de la Recherche Scientifique (CNRS), Laboratoire de Physique Théorique de la Matière Condensée (LPTMC), Centre National de la Recherche Scientifique (CNRS)-Université Pierre et Marie Curie - Paris 6 (UPMC), Laboratoire de Chimie et de Biochimie Pharmacologiques et Toxicologiques (LCBPT - UMR 8601), Université Paris Descartes - Paris 5 (UPD5)-Institut de Chimie du CNRS (INC)-Centre National de la Recherche Scientifique (CNRS), Physiologie intégrative, cellulaire et moléculaire (PICM), Centre National de la Recherche Scientifique (CNRS)-Université Claude Bernard Lyon 1 (UCBL), Université de Lyon-Université de Lyon, HORIBA France SAS [Villeneuve d'Ascq], HORIBA Scientific [France], Centre de biologie du développement (CBD), Centre National de la Recherche Scientifique (CNRS)-Université Toulouse III - Paul Sabatier (UT3), Université Fédérale Toulouse Midi-Pyrénées-Université Fédérale Toulouse Midi-Pyrénées-Centre de Biologie Intégrative (CBI), Université Toulouse III - Paul Sabatier (UT3), Université Fédérale Toulouse Midi-Pyrénées-Université Fédérale Toulouse Midi-Pyrénées-Centre National de la Recherche Scientifique (CNRS)-Centre National de la Recherche Scientifique (CNRS), Surfaces Cellulaires et Signalisation chez les Végétaux (SCSV), Université Fédérale Toulouse Midi-Pyrénées-Université Fédérale Toulouse Midi-Pyrénées, Université Blaise Pascal - Clermont-Ferrand 2 (UBP)-Institut Sciences de l'Homme et de la Société (IR SHS UNILIM), Université de Limoges (UNILIM)-Université de Limoges (UNILIM)-Université Clermont Auvergne [2017-2020] (UCA [2017-2020])-Centre National de la Recherche Scientifique (CNRS)-Université Clermont Auvergne (UCA), Université Claude Bernard Lyon 1 (UCBL), Université de Lyon-Université de Lyon-Centre National de la Recherche Scientifique (CNRS), Université de Toulouse (UT)-Université de Toulouse (UT)-Centre National de la Recherche Scientifique (CNRS)-Centre de Biologie Intégrative (CBI), Université de Toulouse (UT)-Université de Toulouse (UT)-Centre National de la Recherche Scientifique (CNRS)-Centre National de la Recherche Scientifique (CNRS), Université de Toulouse (UT)-Université de Toulouse (UT)-Centre National de la Recherche Scientifique (CNRS), Laboratoire de Physique Théorique des Liquides ( LPTL ), Université Pierre et Marie Curie - Paris 6 ( UPMC ) -Centre National de la Recherche Scientifique ( CNRS ), Laboratoire de Géographie Physique et Environnementale ( GEOLAB ), Centre National de la Recherche Scientifique ( CNRS ) -Institut Sciences de l'Homme et de la Société ( IR SHS UNILIM ), Université de Limoges ( UNILIM ) -Université de Limoges ( UNILIM ) -Institut national de recherches archéologiques préventives ( Inrap ) -Université Blaise Pascal - Clermont-Ferrand 2 ( UBP ) -Université Clermont Auvergne ( UCA ), Laboratoire de Physique Théorique de la Matière Condensée ( LPTMC ), Centre National de la Recherche Scientifique ( CNRS ) -Université Pierre et Marie Curie - Paris 6 ( UPMC ), Laboratoire de Chimie et de Biochimie Pharmacologiques et Toxicologiques ( LCBPT - UMR 8601 ), Université Paris Descartes - Paris 5 ( UPD5 ) -Centre National de la Recherche Scientifique ( CNRS ), Physiologie intégrative, cellulaire et moléculaire ( PICM ), Université Claude Bernard Lyon 1 ( UCBL ), Université de Lyon-Université de Lyon-Centre National de la Recherche Scientifique ( CNRS ), Centre de biologie du développement ( CBD ), Université Toulouse III - Paul Sabatier ( UPS ), Université Fédérale Toulouse Midi-Pyrénées-Université Fédérale Toulouse Midi-Pyrénées-Centre National de la Recherche Scientifique ( CNRS ), Surfaces Cellulaires et Signalisation chez les Végétaux ( SCSV ), Centre National de la Recherche Scientifique (CNRS)-Institut Sciences de l'Homme et de la Société (IR SHS UNILIM), Université de Limoges (UNILIM)-Université de Limoges (UNILIM)-Université Blaise Pascal - Clermont-Ferrand 2 (UBP)-Université Clermont Auvergne (UCA), Université Paris Descartes - Paris 5 (UPD5)-Centre National de la Recherche Scientifique (CNRS), and Université Fédérale Toulouse Midi-Pyrénées-Université Fédérale Toulouse Midi-Pyrénées-Centre National de la Recherche Scientifique (CNRS)

Subjects
MESH : Calcium Signaling, MESH : Gene Expression Regulation, MESH: Calcium, MESH: Animals, [SDV.BBM]Life Sciences [q-bio]/Biochemistry, Molecular Biology, MESH : Animals, MESH : Calcium, MESH: Calcium Signaling, [ SDV.BBM ] Life Sciences [q-bio]/Biochemistry, Molecular Biology, MESH: Gene Expression Regulation
Published
2006

21. Otoferlin, defective in a human deafness form, is essential for exocytosis at the auditory ribbon synapse

Author

Philippe Rostaing, Christine Petit, M'hamed Grati, Ghislaine Hamard, Régis Nouvian, Saaid Safieddine, Marie-Christine Simmler, Amel Bahloul, Antoine Triller, Isabelle Roux, Isabelle Perfettini, Paul Avan, Morgane Le Gall, Tobias Moser, Génétique des Déficits Sensoriels, Institut Pasteur [Paris] (IP)-Institut National de la Santé et de la Recherche Médicale (INSERM), Department of Otolaryngology and Center for Molecular Physiology of the Brain, Georg-August-University = Georg-August-Universität Göttingen, The Wellcome Trust Sanger Institute [Cambridge], Institut Jacques Monod (IJM (UMR_7592)), Université Paris Diderot - Paris 7 (UPD7)-Centre National de la Recherche Scientifique (CNRS), Plateforme Recombinaison Homologue, Transfert d'Embryons et Cryoconservation [Institut Cochin] (PRHTEC), Institut Cochin (IC UM3 (UMR 8104 / U1016)), 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), Biologie cellulaire de la synapse normale et pathologique, Département de Biologie - ENS Paris, École normale supérieure - Paris (ENS-PSL), Université Paris sciences et lettres (PSL)-Université Paris sciences et lettres (PSL)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Centre National de la Recherche Scientifique (CNRS)-École normale supérieure - Paris (ENS-PSL), Université Paris sciences et lettres (PSL)-Université Paris sciences et lettres (PSL)-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), Equipe Biophysique Neurosensorielle [Neuro-Dol], Neuro-Dol (Neuro-Dol), Université d'Auvergne - Clermont-Ferrand I (UdA)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Université d'Auvergne - Clermont-Ferrand I (UdA)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Neuro-Dol (Neuro-Dol), Institut National de la Santé et de la Recherche Médicale (INSERM)-Université Clermont Auvergne [2017-2020] (UCA [2017-2020])-Université Clermont Auvergne [2017-2020] (UCA [2017-2020]), Collège de France - Chaire Génétique et physiologie cellulaire, Collège de France (CdF (institution)), Foundation Louis-Jeantet and the French Ministry of Research (ACI 'Biologie du Développement et Physiologie intégrative') , the European Commission FP6 Integrated Project EuroHear LSHG-CT-2004-512063, Human Frontiers Science Program, the Deutsche Forschungsgemeinschaft, (Center for Molecular Physiology of the Brain), MENRT, Institut Pasteur [Paris]-Institut National de la Santé et de la Recherche Médicale ( INSERM ), Georg-August-Universität Göttingen, Institut Jacques Monod ( IJM ), Université Paris Diderot - Paris 7 ( UPD7 ) -Centre National de la Recherche Scientifique ( CNRS ), Plateforme Recombinaison Homologue, Transfert d'Embryons et Cryoconservation [Institut Cochin] ( PRHTEC ), Institut Cochin ( UM3 (UMR 8104 / U1016) ), 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 ), École normale supérieure - Paris ( ENS Paris ) -Institut National de la Santé et de la Recherche Médicale ( INSERM ), Neuro-Dol ( Neuro-Dol ), Institut National de la Santé et de la Recherche Médicale ( INSERM ) -Université d'Auvergne - Clermont-Ferrand I ( UdA ) -Institut National de la Santé et de la Recherche Médicale ( INSERM ) -Université d'Auvergne - Clermont-Ferrand I ( UdA ) -Neuro-Dol - Clermont Auvergne ( Neuro-Dol ), Institut National de la Santé et de la Recherche Médicale ( INSERM ) -Université Clermont Auvergne ( UCA ) -Université Clermont Auvergne ( UCA ), Institut Pasteur [Paris]-Institut National de la Santé et de la Recherche Médicale (INSERM), Georg-August-University [Göttingen], École normale supérieure - Paris (ENS Paris), Université Paris sciences et lettres (PSL)-Université Paris sciences et lettres (PSL)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Centre National de la Recherche Scientifique (CNRS)-École normale supérieure - Paris (ENS Paris), Chaire Génétique et physiologie cellulaire, Laboratoire d'Electronique et Systèmes de Télécommunications (LEST), Université de Brest (UBO)-Ecole Nationale Supérieure des Télécommunications de Bretagne-Centre National de la Recherche Scientifique (CNRS), Université de Brest (UBO), This work was supported by grants from the FoundationLouis-Jeantet and the French Ministry of Research (ACI ‘‘Biologie duDeveloppement et Physiologie integrative’’) to C.P., Human Frontiers Science Program to T.M. and S.S., and the Deutsche Forschungsgemeinschaft to T.M. (Centerfor Molecular Physiology of the Brain)., École normale supérieure - Paris (ENS Paris)-École normale supérieure - Paris (ENS Paris)-Institut National de la Santé et de la Recherche Médicale (INSERM), Institut National de la Santé et de la Recherche Médicale (INSERM)-Université d'Auvergne - Clermont-Ferrand I (UdA)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Université d'Auvergne - Clermont-Ferrand I (UdA)-Neuro-Dol - Clermont Auvergne (Neuro-Dol), and Institut National de la Santé et de la Recherche Médicale (INSERM)-Université Clermont Auvergne (UCA)-Université Clermont Auvergne (UCA)

Subjects
Auditory Pathways, Time Factors, [SDV]Life Sciences [q-bio], MESH: Synaptosomal-Associated Protein 25, Synaptogenesis, Syntaxin 1, [SDV.BC.BC]Life Sciences [q-bio]/Cellular Biology/Subcellular Processes [q-bio.SC], Ribbon synapse, Deafness, MESH : Synaptosomal-Associated Protein 25, Membrane Fusion, Synaptic Transmission, MESH: Mice, Knockout, MESH: Synapses, Mice, 0302 clinical medicine, MESH: Syntaxin 1, MESH: Cochlea, OTOF, MESH: Animals, Mice, Knockout, 0303 health sciences, MESH : Synaptic Transmission, MESH: Exocytosis, MESH : Hair Cells, Inner, MESH : Evoked Potentials, Auditory, Brain Stem, SNAP25, Anatomy, Cell biology, Cochlea, Synaptic vesicle exocytosis, medicine.anatomical_structure, MESH: Auditory Pathways, MESH: Calcium, MESH : Synaptic Vesicles, Hair cell, Synaptic Vesicles, MESH: Membrane Proteins, MESH : Synapses, MESH : Time Factors, MESH : Cochlea, MESH : Deafness, Synaptosomal-Associated Protein 25, MESH: Deafness, MESH : Mice, Inbred C57BL, Biology, MESH: Membrane Fusion, Synaptic vesicle, General Biochemistry, Genetics and Molecular Biology, Exocytosis, MESH : Exocytosis, 03 medical and health sciences, MESH: Mice, Inbred C57BL, MESH : Membrane Fusion, MESH : Mice, medicine, Evoked Potentials, Auditory, Brain Stem, MESH: Synaptic Transmission, Animals, Humans, MESH : Calcium, MESH: Mice, 030304 developmental biology, Hair Cells, Auditory, Inner, MESH: Humans, Biochemistry, Genetics and Molecular Biology(all), MESH : Humans, MESH: Time Factors, [ SDV.BC.BC ] Life Sciences [q-bio]/Cellular Biology/Subcellular Processes [q-bio.SC], Membrane Proteins, MESH : Syntaxin 1, MESH: Synaptic Vesicles, MESH: Hair Cells, Inner, Mice, Inbred C57BL, MESH : Membrane Proteins, Synapses, MESH : Mice, Knockout, MESH: Evoked Potentials, Auditory, Brain Stem, Calcium, MESH : Animals, MESH : Auditory Pathways, sense organs, 030217 neurology & neurosurgery
Abstract

International audience; The auditory inner hair cell (IHC) ribbon synapse operates with an exceptional temporal precision and maintains a high level of neurotransmitter release. However, the molecular mechanisms underlying IHC synaptic exocytosis are largely unknown. We studied otoferlin, a predicted C2-domain transmembrane protein, which is defective in a recessive form of human deafness. We show that otoferlin expression in the hair cells correlates with afferent synaptogenesis and find that otoferlin localizes to ribbon-associated synaptic vesicles. Otoferlin binds Ca(2+) and displays Ca(2+)-dependent interactions with the SNARE proteins syntaxin1 and SNAP25. Otoferlin deficient mice (Otof(-/-)) are profoundly deaf. Exocytosis in Otof(-/-) IHCs is almost completely abolished, despite normal ribbon synapse morphogenesis and Ca(2+) current. Thus, otoferlin is essential for a late step of synaptic vesicle exocytosis and may act as the major Ca(2+) sensor triggering membrane fusion at the IHC ribbon synapse.

Published
2006
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22. [An increase in intracellular free calcium controls the expression of an arginine N-methyl-transferase involved in neural determination in amphibian embryo]

Author

Néant, Isabelle, Leclerc, Catherine, Batut, Julie, Vandel, Laurence, Moreau, Marc, Centre de biologie du développement (CBD), Centre National de la Recherche Scientifique (CNRS)-Université Toulouse III - Paul Sabatier (UT3), Université Fédérale Toulouse Midi-Pyrénées-Université Fédérale Toulouse Midi-Pyrénées, Laboratoire de Chimie et de Biochimie Pharmacologiques et Toxicologiques (LCBPT - UMR 8601), Université Paris Descartes - Paris 5 (UPD5)-Centre National de la Recherche Scientifique (CNRS), Université Fédérale Toulouse Midi-Pyrénées-Université Fédérale Toulouse Midi-Pyrénées-Centre de Biologie Intégrative (CBI), Université Toulouse III - Paul Sabatier (UT3), Université Fédérale Toulouse Midi-Pyrénées-Université Fédérale Toulouse Midi-Pyrénées-Centre National de la Recherche Scientifique (CNRS)-Centre National de la Recherche Scientifique (CNRS), Université Paris Descartes - Paris 5 (UPD5)-Institut de Chimie du CNRS (INC)-Centre National de la Recherche Scientifique (CNRS), Centre de biologie du développement ( CBD ), Université Toulouse III - Paul Sabatier ( UPS ), Université Fédérale Toulouse Midi-Pyrénées-Université Fédérale Toulouse Midi-Pyrénées-Centre National de la Recherche Scientifique ( CNRS ), Laboratoire de Chimie et de Biochimie Pharmacologiques et Toxicologiques ( LCBPT - UMR 8601 ), and Université Paris Descartes - Paris 5 ( UPD5 ) -Centre National de la Recherche Scientifique ( CNRS )

Subjects
MESH: Cell Differentiation, MESH: Enzyme Induction, MESH : Transcription Factors, MESH: Egtazic Acid, MESH: Bone Morphogenetic Proteins, MESH: Carrier Proteins, MESH: Bone Morphogenetic Protein Receptors, MESH: Calcium Signaling, MESH: Chelating Agents, MESH : Blastula, MESH: Caffeine, MESH : Bone Morphogenetic Protein Receptors, MESH : Bone Morphogenetic Proteins, MESH : Calcium Signaling, MESH: Xenopus laevis, MESH: Gastrula, MESH : Gastrula, MESH: Homeodomain Proteins, MESH: Methyltransferases, MESH : Xenopus laevis, MESH : Chelating Agents, MESH: Animals, [SDV.BBM]Life Sciences [q-bio]/Biochemistry, Molecular Biology, MESH : Homeodomain Proteins, MESH: Blastula, MESH : Calcium, [ SDV.BBM ] Life Sciences [q-bio]/Biochemistry, Molecular Biology, MESH: Xenopus Proteins, MESH : Caffeine, MESH : Methyltransferases, MESH : Organ Culture Techniques, MESH : Ectoderm, MESH: Nervous System, MESH : Enzyme Induction, MESH : Xenopus Proteins, MESH : Egtazic Acid, MESH: Transcription Factors, MESH: Organ Culture Techniques, MESH: Calcium, MESH : Nervous System, MESH : Animals, MESH : Carrier Proteins, MESH : Cell Differentiation, MESH: Ectoderm
Published
2006

23. Nitrate efflux is an essential component of the cryptogein signaling pathway leading to defense responses and hypersensitive cell death in tobacco

Author

David Wendehenne, Alain Pugin, Olivier Lamotte, Jean-Marie Frachisse, Hélène Barbier-Brygoo, Plante - microbe - environnement : biochimie, biologie cellulaire et écologie ( PMEBBCE ), Etablissement National d'Enseignement Supérieur Agronomique de Dijon ( ENESAD ) -Institut National de la Recherche Agronomique ( INRA ) -Université de Bourgogne ( UB ) -Centre National de la Recherche Scientifique ( CNRS ), Institut des sciences du végétal ( ISV ), Centre National de la Recherche Scientifique ( CNRS ), Plante - microbe - environnement : biochimie, biologie cellulaire et écologie (PMEBBCE), Centre National de la Recherche Scientifique (CNRS)-Université de Bourgogne (UB)-Institut National de la Recherche Agronomique (INRA)-Etablissement National d'Enseignement Supérieur Agronomique de Dijon (ENESAD), Institut des sciences du végétal (ISV), Centre National de la Recherche Scientifique (CNRS), and Etablissement National d'Enseignement Supérieur Agronomique de Dijon (ENESAD)-Institut National de la Recherche Agronomique (INRA)-Université de Bourgogne (UB)-Centre National de la Recherche Scientifique (CNRS)

Subjects
0106 biological sciences, MESH: Signal Transduction, MESH: Algal Proteins, MESH: Hydrogen-Ion Concentration, [ SDV.BV ] Life Sciences [q-bio]/Vegetal Biology, MESH : Reactive Oxygen Species, Apoptosis, MESH : Immunity, Natural, Plant Science, 01 natural sciences, Ion Channels, MESH : Phosphorylation, MESH: Plant Diseases, MESH : Anions, Plant defense against herbivory, MESH: Protein Kinase Inhibitors, Phosphorylation, MESH: Tobacco, 0303 health sciences, Fungal protein, MESH : Plant Diseases, food and beverages, MESH: Reactive Oxygen Species, MESH : Nitrates, Hydrogen-Ion Concentration, Cell biology, Elicitor, Respiratory burst, MESH: Nitrates, MESH: Calcium, Natural, Efflux, MESH : Algal Proteins, Signal transduction, MESH : Ion Channels, MESH: Anions, Research Article, Signal Transduction, Anions, Programmed cell death, MESH: Enzyme Activation, MESH : Tobacco, Biology, MESH : Protein Kinases, Fungal Proteins, 03 medical and health sciences, MESH : Hydrogen-Ion Concentration, Tobacco, [SDV.BV]Life Sciences [q-bio]/Vegetal Biology, Protein kinase A, MESH : Calcium, Protein Kinase Inhibitors, MESH: Protein Kinases, 030304 developmental biology, Plant Diseases, MESH: Immunity, Natural, MESH : Signal Transduction, Nitrates, MESH: Phosphorylation, MESH: Apoptosis, Algal Proteins, Immunity, Cell Biology, MESH : Protein Kinase Inhibitors, Immunity, Innate, Enzyme Activation, MESH: Ion Channels, Calcium, Reactive Oxygen Species, MESH : Enzyme Activation, Protein Kinases, MESH : Apoptosis, 010606 plant biology & botany
Abstract

There is much interest in the transduction pathways by which avirulent pathogens or derived elicitors activate plant defense responses. However, little is known about anion channel functions in this process. The aim of this study was to reveal the contribution of anion channels in the defense response triggered in tobacco by the elicitor cryptogein. Cryptogein induced a fast nitrate (NO(3)(-)) efflux that was sensitive to anion channel blockers and regulated by phosphorylation events and Ca(2+) influx. Using a pharmacological approach, we provide evidence that NO(3)(-) efflux acts upstream of the cryptogein-induced oxidative burst and a 40-kD protein kinase whose activation seems to be controlled by the duration and intensity of anion efflux. Moreover, NO(3)(-) efflux inhibitors reduced and delayed the hypersensitive cell death triggered by cryptogein in tobacco plants. This was accompanied by a delay or a complete suppression of the induction of several defense-related genes, including hsr203J, a gene whose expression is correlated strongly with programmed cell death in plants. Our results indicate that anion channels are involved intimately in mediating defense responses and hypersensitive cell death.

Published
2002
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24. Functional antigen-independent synapses formed between T cells and dendritic cells

Author

Mireia Sospedra, Boris Barbour, Alain Trautmann, Patrick Revy, Developpement Normal et Pathologique du Système Immunitaire, Université Paris Descartes - Paris 5 (UPD5)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Centre National de la Recherche Scientifique (CNRS), Neurobiologie cellulaire et moléculaire (NCM), École normale supérieure - Paris (ENS Paris), Université Paris sciences et lettres (PSL)-Université Paris sciences et lettres (PSL)-Centre National de la Recherche Scientifique (CNRS), École normale supérieure - Paris (ENS Paris)-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 ), Neurobiologie cellulaire et moléculaire ( NCM ), and École normale supérieure - Paris ( ENS Paris ) -Centre National de la Recherche Scientifique ( CNRS )

Subjects
T-Lymphocytes, [SDV.NEU.NB]Life Sciences [q-bio]/Neurons and Cognition [q-bio.NC]/Neurobiology, MESH : Antigens, MESH : beta 2-Microglobulin, MESH : Gene Deletion, Lymphocyte Activation, Immunological synapse, MESH : Phosphorylation, Mice, 0302 clinical medicine, Immunology and Allergy, Cytotoxic T cell, MESH: Animals, Phosphorylation, Cells, Cultured, 0303 health sciences, MESH : Cell Survival, MESH: Dendritic Cells, CD28, Natural killer T cell, Cell biology, medicine.anatomical_structure, MESH : Histocompatibility Antigens Class II, MESH: Cell Survival, [ SDV.NEU.NB ] Life Sciences [q-bio]/Neurons and Cognition [q-bio.NC]/Neurobiology, MESH: Calcium, MESH: Antigens, MESH: beta 2-Microglobulin, MESH: Cells, Cultured, Naive T cell, Cell Survival, T cell, Immunology, Antigen presentation, MESH : Mice, Inbred C57BL, Biology, 03 medical and health sciences, MESH: Mice, Inbred C57BL, MESH : Cells, Cultured, MESH : Mice, medicine, Animals, Antigens, Antigen-presenting cell, MESH : Calcium, MESH: Lymphocyte Activation, MESH: Mice, MESH : Lymphocyte Activation, 030304 developmental biology, MESH : T-Lymphocytes, MESH: Phosphorylation, Histocompatibility Antigens Class II, Dendritic Cells, Mice, Inbred C57BL, MESH: T-Lymphocytes, MESH: Gene Deletion, MESH : Dendritic Cells, MESH: Histocompatibility Antigens Class II, Calcium, MESH : Animals, beta 2-Microglobulin, Gene Deletion, 030215 immunology
Abstract

Immunological synapse formation is usually assumed to require antigen recognition by T cell receptors. However, the immunological synapse formed at the interface between naive T cells and dendritic cells (DCs) has never been described. We show here that in the absence of antigen, and even of major histocompatibility complex molecules, T cell-DC synapses are formed and lead to several T cell responses: a local increase in tyrosine phosphorylation, small Ca2+ responses, weak proliferation and long-term survival. These responses are triggered more readily in CD4+ T cells than in CD8+ T cells, which express a specific isoform of the repulsive molecule CD43. These phenomena may play a major role in the maintenance of the naive T cell pool in vivo.

Published
2001
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25. Evidence for an uncommon alpha-actinin protein in Trichomonas vaginalis

Author

Gérard Coffe, Geneviève Bricheux, Guy Brugerolle, Nathalie Pradel, Laboratoire Microorganismes : Génome et Environnement (LMGE), Université Blaise Pascal - Clermont-Ferrand 2 (UBP)-Centre National de la Recherche Scientifique (CNRS)-Université d'Auvergne - Clermont-Ferrand I (UdA), Laboratoire de Biologie des Protistes [Aubière] (LBP), Université Blaise Pascal - Clermont-Ferrand 2 (UBP)-Centre National de la Recherche Scientifique (CNRS), Laboratoire Microorganismes : Génome et Environnement ( LMGE ), Université Blaise Pascal - Clermont-Ferrand 2 ( UBP ) -Université d'Auvergne - Clermont-Ferrand I ( UdA ) -Centre National de la Recherche Scientifique ( CNRS ), BRICHEUX, Genevieve, and Université Blaise Pascal - Clermont-Ferrand 2 (UBP)-Université d'Auvergne - Clermont-Ferrand I (UdA)-Centre National de la Recherche Scientifique (CNRS)

Subjects
MESH: Trichomonas vaginalis, MESH : Molecular Sequence Data, Transcription, Genetic, MESH : DNA, Complementary, Fluorescent Antibody Technique, MESH : Actins, MESH: Protein Structure, Secondary, MESH : Blotting, Western, Actinin, MESH: Amino Acid Sequence, MESH: Base Sequence, Protein Structure, Secondary, Homology (biology), MESH : Cytoskeleton, Protein sequencing, MESH: Actinin, Electrophoresis, Gel, Two-Dimensional, MESH: Animals, Peptide sequence, MESH: Fluorescent Antibody Technique, Cytoskeleton, MESH : Consensus Sequence, MESH : Sequence Alignment, MESH : Amino Acid Sequence, MESH : Blotting, Northern, [ SDV.MP.PRO ] Life Sciences [q-bio]/Microbiology and Parasitology/Protistology, Actinin, alpha 1, Biochemistry, MESH: Calcium, MESH: 5' Untranslated Regions, Electrophoresis, Polyacrylamide Gel, DNA, Complementary, Blotting, Western, Molecular Sequence Data, MESH: Sequence Alignment, Sequence alignment, macromolecular substances, [SDV.MP.PRO] Life Sciences [q-bio]/Microbiology and Parasitology/Protistology, Biology, MESH : Electrophoresis, Polyacrylamide Gel, MESH: Actins, [SDV.MP.PRO]Life Sciences [q-bio]/Microbiology and Parasitology/Protistology, Consensus Sequence, Trichomonas vaginalis, Consensus sequence, MESH: Cytoskeleton, Animals, MESH: Blotting, Northern, MESH: Blotting, Western, Amino Acid Sequence, MESH: Consensus Sequence, MESH : Calcium, Molecular Biology, Actin, MESH: Molecular Sequence Data, Base Sequence, MESH: Transcription, Genetic, MESH : 5' Untranslated Regions, MESH : Transcription, Genetic, MESH : Electrophoresis, Gel, Two-Dimensional, MESH: DNA, Complementary, MESH : Trichomonas vaginalis, Blotting, Northern, MESH: Electrophoresis, Gel, Two-Dimensional, Molecular biology, Actins, MESH : Fluorescent Antibody Technique, Calcium, MESH : Base Sequence, Parasitology, MESH : Protein Structure, Secondary, MESH : Animals, 5' Untranslated Regions, Sequence Alignment, MESH : Actinin, MESH: Electrophoresis, Polyacrylamide Gel
Abstract

International audience; As part of our ongoing project of identification of actin-binding proteins implicated in the cell transition (flagellate to amoeboid/adherent) of Trichomonas vaginalis, we have characterized an alpha-actinin-related protein in this parasite. The protein (P100) has a molecular mass of 100 kDa and an isoelectric point of 5.5. A monoclonal antibody raised against this protein co-localizes with the actin network. P100 gene transcripts are co-expressed with actin throughout the cell cycle. Analysis of the deduced protein sequence reveals three domains: an N-terminal actin-binding region; a central region rich in alpha-helix; and a C-terminal domain with Ca(2+)-binding capacity. Whereas the N- and C-terminal regions are well-conserved as compared to other alpha-actinins, we observe in the central region an atypical distribution of residues in five repeats. The sequence of the repeats does not show any homology with the rod domain of the other alpha-actinins, except for the first repeat which shows some similarity. The four other repeats of T. vaginalis P100 appear to result from a duplication event which is not detectable in the other sequences.

Published
1998

26. Thyroid hormones and the creatine kinase system in cardiac cells

Author

Enn Seppet, Valdur Saks, Hamant, Sarah, Department of Pathophysiology, University of Tartu, Institute of Experimental Cardiology, Cardiology Research Center, Laboratory of Molecular Biochemistry, and Russian Academy of Medical Sciences

Subjects
Phosphocreatine, endocrine system diseases, Clinical Biochemistry, Creatine transport, MESH : Creatine Kinase, Hyperthyroidism, Oxidative Phosphorylation, chemistry.chemical_compound, MESH : Myocardium, Creatine Kinase, biology, Thyroid, Heart, General Medicine, MESH : Thyroid Hormones, medicine.anatomical_structure, MESH : Oxidative Phosphorylation, Intracellular, Thyroid Hormones, endocrine system, medicine.medical_specialty, Oxidative phosphorylation, In Vitro Techniques, Creatine, MESH : Heart, Hypothyroidism, Internal medicine, MESH : Hyperthyroidism, medicine, [SDV.BBM] Life Sciences [q-bio]/Biochemistry, Molecular Biology, Animals, Humans, MESH : Phosphocreatine, Kinase activity, MESH : Calcium, MESH : Hypothyroidism, [ SDV.BBM ] Life Sciences [q-bio]/Biochemistry, Molecular Biology, Molecular Biology, Myocardium, MESH : Humans, Cell Biology, MESH : Energy Metabolism, Endocrinology, chemistry, biology.protein, Calcium, Creatine kinase, MESH : Animals, Energy Metabolism
Abstract

International audience; The paper reviews the current evidence on the role of thyroid hormones in regulating the creatine kinase energy transfer system at multiple structures in cardiac cells. 1) Thyroid hormones modulate the overall synthesis of phosphocreatine (PCr) by increasing the rate of mitochondrial oxidative phosphorylation. 2) Thyroid hormones regulate the total activity of creatine kinase and its isoenzyme distribution. In comparison with normal thyroid state (euthyroidism), hypothyroidism is characterized by decreased total creatine kinase activity owing to diminished fraction of creatine kinase. On the other hand, hyperthyroidism, while causing no change in total creatine kinase activity, leads to increased fractions of neonatal isoforms of creatine kinase, and, in case of prolonged hyperthyroidism, to decreased fraction of mitochondrial creatine kinase. The latter change is associated with partial uncoupling between mitochondrial creatine kinase and adenine nucleotide translocase reflected by decreased PCr/O ratio. 3) Hyperthyroidism leads to increased passive sarcolemmal permeability due to which the leakage of creatine along its concentration gradient occurs. As a result of (i) increased sarcolemmal permeability for creatine, (ii) uncoupling of mitochondrial PCr synthesis, and (iii) increased energy utilization rate the steady state intracellular PCr content decreases under hyperthyroidism which, in turn, increases the myocardial susceptibility to hypoxic damage. Thyroid state also modulates the protective effects of exogenous PCr on energetically depleted myocardium.

Published
1994

27. ACE inhibition prevents diastolic Ca2+ overload and loss of myofilament Ca2+ sensitivity after myocardial infarction. : Molecular effects of delapril on cardiomyocytes

Author

Christophe Heymes, Lucas Andre, Ivar Sjaastad, Cécile Cassan, Franck Aimond, Sylvain Richard, Jean-Luc Pasquié, Yannis Sainte-Marie, Olivier Cazorla, Jérôme Thireau, Santiago Zalvidea, Xavier Loyer, Physiologie & médecine expérimentale du Cœur et des Muscles [U 1046] ( PhyMedExp ), Institut National de la Santé et de la Recherche Médicale ( INSERM ) -Université de Montpellier ( UM ) -Centre National de la Recherche Scientifique ( CNRS ), Paris-Centre de Recherche Cardiovasculaire ( PARCC - U970 ), Université Paris Descartes - Paris 5 ( UPD5 ) -Institut National de la Santé et de la Recherche Médicale ( INSERM ) -Hôpital Européen Georges Pompidou [APHP] ( HEGP ), Institut des Maladies Métaboliques et Cardiovasculaires ( I2MC ), Institut National de la Santé et de la Recherche Médicale ( INSERM ) -Hôpital de Rangueil, CHU Toulouse [Toulouse]-CHU Toulouse [Toulouse]-Université Toulouse III - Paul Sabatier ( UPS ), Université Fédérale Toulouse Midi-Pyrénées-Université Fédérale Toulouse Midi-Pyrénées, Institute for Experimental Medical Research, and Oslo University Hospital

Subjects
Male, Myocardial Infarction, heart failure, 030204 cardiovascular system & hematology, Biochemistry, Muscle hypertrophy, Mice, 0302 clinical medicine, Myofibrils, Diastole, Troponin I, Medicine, Myocardial infarction, Excitation Contraction Coupling, MESH : Myocardial Contraction, myofilaments, MESH : Ryanodine Receptor Calcium Release Channel, 0303 health sciences, Ventricular Remodeling, excitation-contraction coupling, sarcoplasmic reticulum Ca2+ ATPase, General Medicine, [ SDV.MHEP.CSC ] Life Sciences [q-bio]/Human health and pathology/Cardiology and cardiovascular system, 3. Good health, Sarcoplasmic Reticulum, Cardiology, Molecular Medicine, MESH : Angiotensin-Converting Enzyme Inhibitors, MESH : Sarcoplasmic Reticulum, hypertrophy, MESH : Ventricular Remodeling, medicine.drug, Cardiac function curve, medicine.medical_specialty, MESH : Male, Delapril, Article, 03 medical and health sciences, MESH : Excitation Contraction Coupling, Internal medicine, Angiotensin-converting enzyme inhibitors, MESH : Mice, Animals, Ventricular remodeling, MESH : Calcium, Molecular Biology, 030304 developmental biology, business.industry, MESH : Diastole, Ryanodine Receptor Calcium Release Channel, medicine.disease, Myocardial Contraction, MESH : Myofibrils, MESH : Disease Models, Animal, Disease Models, Animal, Endocrinology, Heart failure, Calcium, MESH : Animals, MESH : Myocardial Infarction, business
Abstract

International audience; Prevention of adverse cardiac remodeling after myocardial infarction (MI) remains a therapeutic challenge. Angiotensin-converting enzyme inhibitors (ACE-I) are a well-established first-line treatment. ACE-I delay fibrosis, but little is known about their molecular effects on cardiomyocytes. We investigated the effects of the ACE-I delapril on cardiomyocytes in a mouse model of heart failure (HF) after MI. Mice were randomly assigned to three groups: Sham, MI, and MI-D (6 weeks of treatment with a non-hypotensive dose of delapril started 24h after MI). Echocardiography and pressure-volume loops revealed that MI induced hypertrophy and dilation, and altered both contraction and relaxation of the left ventricle. At the cellular level, MI cardiomyocytes exhibited reduced contraction, slowed relaxation, increased diastolic Ca2+ levels, decreased Ca2+-transient amplitude, and diminished Ca2+ sensitivity of myofilaments. In MI-D mice, however, both mortality and cardiac remodeling were decreased when compared to non-treated MI mice. Delapril maintained cardiomyocyte contraction and relaxation, prevented diastolic Ca2+ overload and retained the normal Ca2+ sensitivity of contractile proteins. Delapril maintained SERCA2a activity through normalization of P-PLB/PLB (for both Ser16- PLB and Thr17-PLB) and PLB/SERCA2a ratios in cardiomyocytes, favoring normal reuptake of Ca2+ in the sarcoplasmic reticulum. In addition, delapril prevented defective cTnI function by normalizing the expression of PKC, enhanced in MI mice. In conclusion, early therapy with delapril after MI preserved the normal contraction/relaxation cycle of surviving cardiomyocytes with multiple direct effects on key intracellular mechanisms contributing to preserve cardiac function.

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