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23 results on '"Biologie des Jonctions Neuromusculaires Normales et Pathologiques ( U686 )"'

1. Identification of an Agrin Mutation that Causes Congenital Myasthenia and Affects Synapse Function

Author

Michel Fardeau, Laurent Schaeffer, Thierry Kuntzer, Isabelle Grosjean, Daniel Hantaï, A. Rouche, Annie Chaboud, Asma Ben Ammar, Nektaria Alexandri, Frédéric Chevessier, Caroline Huzé, Emmanuel Fournier, Andrea Brancaccio, K. Gaudon, Bruno Eymard, Jeanine Koenig, S. Bauche, Evelyne Goillot, Véronique Bernard, Heba-Aude Lecuyer, Pascale Richard, Markus A. Rüegg, 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 ), Centre de Recherche de l'Institut du Cerveau et de la Moelle épinière ( CRICM ), Université Pierre et Marie Curie - Paris 6 ( UPMC ) -Institut National de la Santé et de la Recherche Médicale ( INSERM ) -Centre National de la Recherche Scientifique ( CNRS ), Unité Fonctionnelle de Cardiogénétique et Myogénétique Moléculaire et Cellulaire, Assistance publique - Hôpitaux de Paris (AP-HP)-Centre de Génétique Moléculaire et Chromosomique du GH Pitié-Salpêtrière-CHU Pitié-Salpêtrière [APHP], Max-Planck-Institut für Medizinische Forschung, Max-Planck-Gesellschaft, Institut National de Neurologie, Université Tunis El Manar ( UTM ), Plateau d'analyse des protéines, IFR128, Biologie des Jonctions Neuromusculaires Normales et Pathologiques ( U686 ), Université Paris Descartes - Paris 5 ( UPD5 ) -Institut National de la Santé et de la Recherche Médicale ( INSERM ) -Centre National de la Recherche Scientifique ( CNRS ), Nerve-muscle unit, neurology service, Université de Lausanne ( UNIL ) -Centre Hospitalier Universitaire Vaudois [Lausanne] ( CHUV ), Centre de référence des maladies rares neuromusculaires, Assistance publique - Hôpitaux de Paris (AP-HP)-CHU Pitié-Salpêtrière [APHP], Service d'électrophysiologie, Istituto di Chimica del Riconoscimento Molecolare, Università Cattolica del Sacro Cuore, Biozentrum, University of Basel ( Unibas ), ANR-07-MRAR-0001,MRAR,Syndromes myasthéniques congénitaux : le réseau français et approches fondamentales ( 2007 ), Hantaï, Daniel, Programme Pluriannuel de Recherche sur les Maladies Rares (MRAR) - Implication de CXCL13 et CCL21 dans les mécanismes pathogéniques de la Myasthenie. - - MG chemokines2006 - ANR-06-MRAR-0001 - MRAR - VALID, 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), Centre de Recherche de l'Institut du Cerveau et de la Moelle épinière (CRICM), Université Pierre et Marie Curie - Paris 6 (UPMC)-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)-CHU Pitié-Salpêtrière [AP-HP], Assistance publique - Hôpitaux de Paris (AP-HP) (AP-HP)-Sorbonne Université (SU)-Sorbonne Université (SU), Université de Tunis El Manar (UTM), Biologie des Jonctions Neuromusculaires Normales et Pathologiques (U686), Université Paris Descartes - Paris 5 (UPD5)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Centre National de la Recherche Scientifique (CNRS), Neuroscience Paris Seine (NPS), Université Pierre et Marie Curie - Paris 6 (UPMC)-Institut de Biologie Paris Seine (IBPS), Université Pierre et Marie Curie - Paris 6 (UPMC)-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 National de la Recherche Scientifique (CNRS)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Centre National de la Recherche Scientifique (CNRS), Université de Lausanne = University of Lausanne (UNIL)-Centre Hospitalier Universitaire Vaudois [Lausanne] (CHUV), Centre de référence des maladies rares neuromusculaires [CHU Pitié-Salpétriêre], CHU Pitié-Salpêtrière [AP-HP], Assistance publique - Hôpitaux de Paris (AP-HP) (AP-HP)-Sorbonne Université (SU)-Assistance publique - Hôpitaux de Paris (AP-HP) (AP-HP)-Sorbonne Université (SU), Assistance publique - Hôpitaux de Paris (AP-HP) (AP-HP)-Sorbonne Université (SU), Università cattolica del Sacro Cuore [Milano] (Unicatt), Biozentrum [Basel, Suisse], University of Basel (Unibas), ANR-06-MRAR-0001,MG chemokines,Implication de CXCL13 et CCL21 dans les mécanismes pathogéniques de la Myasthenie.(2006), École normale supérieure de Lyon (ENS de Lyon)-Université Claude Bernard Lyon 1 (UCBL), 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), Sorbonne Université (SU)-Assistance publique - Hôpitaux de Paris (AP-HP) (AP-HP)-Sorbonne Université (SU), Neurosciences Paris Seine (NPS), Université Pierre et Marie Curie - Paris 6 (UPMC)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Institut de Biologie Paris Seine (IBPS), Université Pierre et Marie Curie - Paris 6 (UPMC)-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 National de la Recherche Scientifique (CNRS)-Centre National de la Recherche Scientifique (CNRS), Centre Hospitalier Universitaire Vaudois [Lausanne] (CHUV)-Université de Lausanne (UNIL), Sorbonne Université (SU)-Assistance publique - Hôpitaux de Paris (AP-HP) (AP-HP), Sorbonne Université (SU)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Institut de Biologie Paris Seine (IBPS), Sorbonne Université (SU)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Centre National de la Recherche Scientifique (CNRS)-Sorbonne Université (SU)-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 du Cerveau et de la Moëlle Epinière = Brain and Spine Institute (ICM), Université Pierre et Marie Curie - Paris 6 (UPMC)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Centre National de la Recherche Scientifique (CNRS)-CHU Pitié-Salpêtrière [AP-HP], Sorbonne Université-Assistance publique - Hôpitaux de Paris (AP-HP) (AP-HP)-Sorbonne Université-Assistance publique - Hôpitaux de Paris (AP-HP) (AP-HP), Sorbonne Université-Assistance publique - Hôpitaux de Paris (AP-HP) (AP-HP)-Sorbonne Université, Institut National de la Santé et de la Recherche Médicale (INSERM)-Sorbonne Université (SU)-Centre National de la Recherche Scientifique (CNRS)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Sorbonne Université (SU)-Centre National de la Recherche Scientifique (CNRS)-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)-CHU Pitié-Salpêtrière [AP-HP], and Sorbonne Université-Assistance publique - Hôpitaux de Paris (AP-HP) (AP-HP)-Sorbonne Université-Assistance publique - Hôpitaux de Paris (AP-HP) (AP-HP)-Centre National de la Recherche Scientifique (CNRS)

Subjects
Male, Biopsy, DNA Mutational Analysis, Muscle Fibers, Skeletal, Mutant, [SDV.GEN] Life Sciences [q-bio]/Genetics, 0302 clinical medicine, Mutant protein, CHRNE, Receptors, Cholinergic, Genetics(clinical), Dystroglycans, ComputingMilieux_MISCELLANEOUS, Genetics (clinical), Genetics, 0303 health sciences, Agrin, Congenital myasthenic syndrome, Recombinant Proteins, Pedigree, Cell biology, medicine.anatomical_structure, Female, [SDV.NEU]Life Sciences [q-bio]/Neurons and Cognition [q-bio.NC], Dok-7, Adult, animal structures, Mutation, Missense, Neuromuscular Junction, Biology, Article, Neuromuscular junction, Cell Line, 03 medical and health sciences, medicine, Animals, Humans, [SDV.NEU] Life Sciences [q-bio]/Neurons and Cognition [q-bio.NC], Muscle, Skeletal, 030304 developmental biology, Acetylcholine receptor, Myasthenic Syndromes, Congenital, [SDV.GEN]Life Sciences [q-bio]/Genetics, Correction, medicine.disease, Protein Structure, Tertiary, Rats, Models, Chemical, [ SDV.NEU ] Life Sciences [q-bio]/Neurons and Cognition [q-bio.NC], Synapses, 030221 ophthalmology & optometry, biology.protein, Agrin/chemistry, Agrin/genetics, Dystroglycans/metabolism, Muscle Fibers, Skeletal/cytology, Muscle Fibers, Skeletal/metabolism, Muscle, Skeletal/metabolism, Muscle, Skeletal/pathology, Myasthenic Syndromes, Congenital/genetics, Neuromuscular Junction/genetics, Neuromuscular Junction/metabolism, Receptors, Cholinergic/genetics, Receptors, Cholinergic/metabolism, Recombinant Proteins/chemistry, Recombinant Proteins/metabolism, Synapses/metabolism, [ SDV.GEN ] Life Sciences [q-bio]/Genetics, 030217 neurology & neurosurgery
Abstract

International audience; We report the case of a congenital myasthenic syndrome due to a mutation in AGRN, the gene encoding agrin, an extracellular matrix molecule released by the nerve and critical for formation of the neuromuscular junction. Gene analysis identified a homozygous missense mutation, c.5125G>C, leading to the p.Gly1709Arg variant. The muscle-biopsy specimen showed a major disorganization of the neuromuscular junction, including changes in the nerve-terminal cytoskeleton and fragmentation of the synaptic gutters. Experiments performed in nonmuscle cells or in cultured C2C12 myotubes and using recombinant mini-agrin for the mutated and the wild-type forms showed that the mutated form did not impair the activation of MuSK or change the total number of induced acetylcholine receptor aggregates. A solid-phase assay using the dystrophin glycoprotein complex showed that the mutation did not affect the binding of agrin to alpha-dystroglycan. Injection of wild-type or mutated agrin into rat soleus muscle induced the formation of nonsynaptic acetylcholine receptor clusters, but the mutant protein specifically destabilized the endogenous neuromuscular junctions. Importantly, the changes observed in rat muscle injected with mutant agrin recapitulated the pre- and post-synaptic modifications observed in the patient. These results indicate that the mutation does not interfere with the ability of agrin to induce postsynaptic structures but that it dramatically perturbs the maintenance of the neuromuscular junction.

Published
2009
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2. MuSK frizzled-like domain is critical for mammalian neuromuscular junction formation and maintenance

Author

Julien Messéant, Marin Manuel, Daniel Zytnicki, Alexandre Dobbertin, Laurent Schaeffer, Claire Legay, Alain Schmitt, Perrine Delers, Emmanuelle Girard, Francesca Mangione, Laure Strochlic, Dominique Le Denmat, Jordi Molgó, Centre de neurophysique, physiologie, pathologie (UMR 8119), Université Paris Descartes - Paris 5 (UPD5)-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), Pathologies, Imagerie et Biothérapies oro-faciales (EA 2496), Université Paris Descartes - Paris 5 (UPD5), 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), Institut des Neurosciences de Paris-Saclay (Neuro-PSI), Université Paris-Sud - Paris 11 (UP11)-Centre National de la Recherche Scientifique (CNRS), Neurophysique et physiologie du système moteur (NPSM), Biologie des Jonctions Neuromusculaires Normales et Pathologiques (U686), École normale supérieure de Lyon (ENS de Lyon)-Université Claude Bernard Lyon 1 (UCBL), Institut des Neurosciences Paris-Saclay (NeuroPSI), Association Francaise contre les Myopathies (14960, 18046), Centre National de la Recherche Scientifique, Paris Descartes University, Institut National de la Sante et de la Recherche Medicale, Institut National de la Santé et de la Recherche Médicale (INSERM)-Centre National de la Recherche Scientifique (CNRS)-École normale supérieure - Lyon (ENS Lyon)-Université Claude Bernard Lyon 1 (UCBL), Université de Lyon-Université de Lyon, 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 de neurophysique, physiologie, pathologie ( UMR 8119 ), Université Paris Descartes - Paris 5 ( UPD5 ) -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 ), Pathologies, Imagerie et Biothérapies oro-faciales ( EA 2496 ), Université Paris Descartes - Paris 5 ( UPD5 ), 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 ), Institut des Neurosciences de Paris-Saclay ( Neuro-PSI ), Université Paris-Sud - Paris 11 ( UP11 ) -Centre National de la Recherche Scientifique ( CNRS ), Neurophysique et physiologie du système moteur ( NPSM ), and Biologie des Jonctions Neuromusculaires Normales et Pathologiques ( U686 )

Subjects
Male, MESH: Fatigue, Frizzled, [SDV.NEU.NB]Life Sciences [q-bio]/Neurons and Cognition [q-bio.NC]/Neurobiology, MESH : Muscle Weakness, Synaptogenesis, MESH: Animals, Newborn, Synapse, Mice, [ SDV.NEU.SC ] Life Sciences [q-bio]/Neurons and Cognition [q-bio.NC]/Cognitive Sciences, 0302 clinical medicine, MESH: Pregnancy, Pregnancy, Receptors, Cholinergic, MESH : Female, MESH: Animals, Fatigue, MuSK, Motor Neurons, 0303 health sciences, Muscle Weakness, synaptogenesis, Hand Strength, neuromuscular junction, [SDV.NEU.PC]Life Sciences [q-bio]/Neurons and Cognition [q-bio.NC]/Psychology and behavior, MESH : Animals, Newborn, General Neuroscience, Intracellular Signaling Peptides and Proteins, Wnt signaling pathway, MESH: Muscle Weakness, [SDV.NEU.SC]Life Sciences [q-bio]/Neurons and Cognition [q-bio.NC]/Cognitive Sciences, MESH: Lithium Chloride, Articles, Congenital myasthenic syndrome, MESH : Mice, Transgenic, MESH : Fatigue, medicine.anatomical_structure, MESH: Hand Strength, [ SDV.NEU.NB ] Life Sciences [q-bio]/Neurons and Cognition [q-bio.NC]/Neurobiology, Acetylcholinesterase, Female, MESH: Receptor Protein-Tyrosine Kinases, MESH : Mutation, medicine.symptom, MESH : Lithium Chloride, MESH : Hand Strength, MESH: Motor Neurons, MESH: Myasthenic Syndromes, Congenital, animal structures, MESH: Mutation, MESH : Motor Neurons, MESH: Mice, Transgenic, MESH : Male, Primary Cell Culture, MESH: Glycoproteins, Mice, Transgenic, Biology, MESH : Myasthenic Syndromes, Congenital, Neuromuscular junction, MESH : Acetylcholinesterase, [ SDV.NEU.PC ] Life Sciences [q-bio]/Neurons and Cognition [q-bio.NC]/Psychology and behavior, MESH: Primary Cell Culture, 03 medical and health sciences, Wnt, MESH : Mice, medicine, Animals, MESH : Primary Cell Culture, MESH: Mice, lithium chloride, Glycoproteins, 030304 developmental biology, Acetylcholine receptor, Myasthenic Syndromes, Congenital, MESH: Receptors, Cholinergic, Receptor Protein-Tyrosine Kinases, Muscle weakness, MESH : Receptor Protein-Tyrosine Kinases, MESH: Acetylcholinesterase, MESH : Receptors, Cholinergic, medicine.disease, MESH : Glycoproteins, MESH: Male, MESH : Pregnancy, Animals, Newborn, nervous system, congenital myasthenic syndrome, Mutation, MESH : Neuromuscular Junction, MESH : Animals, MESH: Neuromuscular Junction, Neuroscience, MESH: Female, 030217 neurology & neurosurgery
Abstract

The muscle-specific kinase MuSK is one of the key molecules orchestrating neuromuscular junction (NMJ) formation. MuSK interacts with the Wnt morphogens, through its Frizzled-like domain (cysteine-rich domain [CRD]). Dysfunction of MuSK CRD in patients has been recently associated with the onset of myasthenia, common neuromuscular disorders mainly characterized by fatigable muscle weakness. However, the physiological role of Wnt-MuSK interaction in NMJ formation and function remains to be elucidated. Here, we demonstrate that the CRD deletion of MuSK in mice caused profound defects of both muscle prepatterning, the first step of NMJ formation, and synapse differentiation associated with a drastic deficit in AChR clusters and excessive growth of motor axons that bypass AChR clusters. Moreover, adultMuSKΔCRDmice developed signs of congenital myasthenia, including severe NMJs dismantlement, muscle weakness, and fatigability. We also report, for the first time, the beneficial effects of lithium chloride, a reversible inhibitor of the glycogen synthase kinase-3, that rescued NMJ defects inMuSKΔCRDmice and therefore constitutes a novel therapeutic reagent for the treatment of neuromuscular disorders linked to Wnt-MuSK signaling pathway deficiency. Together, our data reveal that MuSK CRD is critical for NMJ formation and plays an unsuspected role in NMJ maintenance in adulthood.

Published
2015
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3. A mouse model for congenital myasthenic syndrome due to MuSK mutations reveals defects in structure and function of neuromuscular junctions

Author

Sönke Bartling, Jeanine Koenig, Daniel Hantaï, Frédéric Chevessier, Emmanuelle Girard, Veit Witzemann, Jordi Molgó, Max-Planck-Institut für Medizinische Forschung, Max-Planck-Gesellschaft, Biologie des Jonctions Neuromusculaires Normales et Pathologiques (U686), 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 de neurobiologie cellulaire et moléculaire (NBCM), Centre National de la Recherche Scientifique (CNRS), Institut de Neurobiologie Alfred Fessard (INAF), Abteilung medizinische Physik in der Radiologie, Deutsches Krebsforschungzentrum, Physiopathologie et thérapie du muscle strié, Université Pierre et Marie Curie - Paris 6 (UPMC)-IFR14-Institut National de la Santé et de la Recherche Médicale (INSERM), Université Bordeaux Segalen - Bordeaux 2, Centre de référence des maladies neuromusculaires Paris-Est, Hôpital de la Salpêtrière AP-HP, CHU Pitié-Salpêtrière [AP-HP], Assistance publique - Hôpitaux de Paris (AP-HP) (AP-HP)-Sorbonne Université (SU), Biologie des Jonctions Neuromusculaires Normales et Pathologiques ( U686 ), 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 de neurobiologie cellulaire et moléculaire ( NBCM ), Centre National de la Recherche Scientifique ( CNRS ), Institut de Neurobiologie Alfred Fessard ( INAF ), and Université Pierre et Marie Curie - Paris 6 ( UPMC ) -IFR14-Institut National de la Santé et de la Recherche Médicale ( INSERM )

Subjects
Male, Refractory Period, Electrophysiological, MESH: Muscle Contraction, MESH : Muscle Weakness, medicine.disease_cause, Synaptic Transmission, MESH: Motor Endplate, Mice, MESH : Locomotion, 0302 clinical medicine, Missense mutation, MESH : Female, Receptors, Cholinergic, MESH: Animals, Genetics (clinical), MESH : Synaptic Transmission, 0303 health sciences, Mutation, Muscle Weakness, MESH: Muscle Weakness, General Medicine, Congenital myasthenic syndrome, Null allele, medicine.anatomical_structure, Female, MESH: Receptor Protein-Tyrosine Kinases, [SDV.NEU]Life Sciences [q-bio]/Neurons and Cognition [q-bio.NC], medicine.symptom, MESH: Refractory Period, Electrophysiological, Locomotion, Muscle Contraction, Muscle contraction, medicine.medical_specialty, MESH: Axons, MESH: Myasthenic Syndromes, Congenital, MESH : Male, Diaphragm, MESH : Axons, Mutation, Missense, Neuromuscular Junction, MESH: Locomotion, MESH: Microscopy, Electron, Biology, MESH : Myasthenic Syndromes, Congenital, Motor Endplate, Neuromuscular junction, 03 medical and health sciences, Internal medicine, MESH : Mice, Genetics, medicine, MESH: Synaptic Transmission, Animals, Humans, Kyphosis, Molecular Biology, MESH: Mice, 030304 developmental biology, Myasthenic Syndromes, Congenital, MESH: Mutation, Missense, MESH : Kyphosis, MESH: Receptors, Cholinergic, MESH: Humans, MESH : Humans, MESH : Motor Endplate, Receptor Protein-Tyrosine Kinases, Muscle weakness, MESH : Receptor Protein-Tyrosine Kinases, MESH : Receptors, Cholinergic, medicine.disease, MESH : Disease Models, Animal, MESH : Microscopy, Electron, Axons, Myasthenia gravis, MESH: Male, Disease Models, Animal, Microscopy, Electron, MESH : Refractory Period, Electrophysiological, Endocrinology, MESH: Diaphragm, [ SDV.NEU ] Life Sciences [q-bio]/Neurons and Cognition [q-bio.NC], MESH : Muscle Contraction, MESH : Neuromuscular Junction, MESH : Animals, MESH: Neuromuscular Junction, MESH : Diaphragm, MESH: Disease Models, Animal, MESH: Kyphosis, MESH: Female, MESH : Mutation, Missense, 030217 neurology & neurosurgery
Abstract

International audience; In the muscle-specific tyrosine kinase receptor gene MUSK, a heteroallelic missense and a null mutation were identified in a patient suffering from a congenital myasthenic syndrome (CMS). We generated one mouse line carrying the homozygous missense mutation V789M in musk (musk(V789M/V789M) mice) and a second hemizygous line, resembling the patient genotype, with the V789M mutation on one allele and an allele lacking the kinase domain (musk(V789M/-) mice). We report here that musk(V789M/V789M) mice present no obvious abnormal phenotype regarding weight, muscle function and viability. In contrast, adult musk(V789M/-) mice suffer from severe muscle weakness, exhibit shrinkage of pelvic and scapular regions and hunchback. Musk(V789M/-) diaphragm develops less force upon direct or nerve-induced stimulation. A profound tetanic fade is observed following nerve-evoked muscle contraction, and fatigue resistance is severely impaired upon a train of tetanic nerve stimulations. Electrophysiological measurements indicate that fatigable muscle weakness is due to impaired neurotransmission as observed in a patient suffering from a CMS. The diaphragm of adult musk(V789M/-) mice exhibits pronounced changes in endplate architecture, distribution and innervation pattern. Thus, the missense mutation V789M in MuSK acts as a hypomorphic mutation and leads to insufficiency in MuSK function in musk(V789M/-) mutants. These mutant mice represent valuable models for elucidating the roles of MuSK for synapse formation, maturation and maintenance as well as for studying the pathophysiology of a CMS due to MuSK mutations.

Published
2008
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4. Evidence of a dosage effect and a physiological endplate acetylcholinesterase deficiency in the first mouse models mimicking Schwartz-Jampel syndrome neuromyotonia

Author

Morgane Stum, Arnaud Ferry, Frédédrique Rene, Christophe Marcel, Andoni Echaniz-Laguna, Véronique Bernard, Claire-Sophie Davoine, Bertrand Fontaine, Eric Krejci, Sophie Nicole, Alban Vignaud, Marie Bangratz, Emmanuelle Girard, Jordi Molgó, Marc Herbin, Affections de la Myeline et des Canaux Ioniques Musculaires, Université Pierre et Marie Curie - Paris 6 (UPMC)-Institut National de la Santé et de la Recherche Médicale (INSERM), Laboratoire de neurobiologie cellulaire et moléculaire (NBCM), Centre National de la Recherche Scientifique (CNRS), Institut de Neurobiologie Alfred Fessard (INAF), Biologie des Jonctions Neuromusculaires Normales et Pathologiques (U686), Université Paris Descartes - Paris 5 (UPD5)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Centre National de la Recherche Scientifique (CNRS), Mécanismes adaptatifs : des organismes aux communautés (MAOAC), Muséum national d'Histoire naturelle (MNHN)-Université Pierre et Marie Curie - Paris 6 (UPMC)-Centre National de la Recherche Scientifique (CNRS), Groupe Myologie, Institut de Myologie, Université Pierre et Marie Curie - Paris 6 (UPMC)-Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Assistance publique - Hôpitaux de Paris (AP-HP) (AP-HP)-Association française contre les myopathies (AFM-Téléthon)-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)-Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Assistance publique - Hôpitaux de Paris (AP-HP) (AP-HP)-Association française contre les myopathies (AFM-Téléthon)-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), Laboratoire de signalisation moléculaire et neurodégénerescence, Université Louis Pasteur - Strasbourg I-IFR37-Institut National de la Santé et de la Recherche Médicale (INSERM), Département de Neurologie, Hôpital Civil de Strasbourg, CHU Pitié-Salpêtrière [AP-HP], Assistance publique - Hôpitaux de Paris (AP-HP) (AP-HP)-Sorbonne Université (SU), Fédération des Maladies du Système Nerveux, Assistance publique - Hôpitaux de Paris (AP-HP) (AP-HP)-CHU Pitié-Salpêtrière [AP-HP], Sorbonne Université (SU)-Assistance publique - Hôpitaux de Paris (AP-HP) (AP-HP)-Sorbonne Université (SU), Institut National de la Santé et de la Recherche Médicale (INSERM)-Université Pierre et Marie Curie - Paris 6 (UPMC), Muséum national d'Histoire naturelle (MNHN)-Collège de France (CdF)-Centre National de la Recherche Scientifique (CNRS), Université Pierre et Marie Curie - Paris 6 (UPMC)-Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Assistance publique - Hôpitaux de Paris (AP-HP) (APHP)-Association française contre les myopathies (AFM-Téléthon)-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)-Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Assistance publique - Hôpitaux de Paris (AP-HP) (APHP)-Association française contre les myopathies (AFM-Téléthon)-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), Assistance publique - Hôpitaux de Paris (AP-HP) (APHP)-CHU Pitié-Salpêtrière [APHP], Institut National de la Santé et de la Recherche Médicale ( INSERM ) -Université Pierre et Marie Curie - Paris 6 ( UPMC ), Laboratoire de neurobiologie cellulaire et moléculaire ( NBCM ), Centre National de la Recherche Scientifique ( CNRS ), Institut de Neurobiologie Alfred Fessard ( INAF ), Biologie des Jonctions Neuromusculaires Normales et Pathologiques ( U686 ), Université Paris Descartes - Paris 5 ( UPD5 ) -Institut National de la Santé et de la Recherche Médicale ( INSERM ) -Centre National de la Recherche Scientifique ( CNRS ), Mécanismes adaptatifs : des organismes aux communautés ( MAOAC ), Muséum National d'Histoire Naturelle ( MNHN ) -Collège de France ( CdF ) -Centre National de la Recherche Scientifique ( CNRS ), Université Pierre et Marie Curie - Paris 6 ( UPMC ) -Commissariat à l'énergie atomique et aux énergies alternatives ( CEA ) -Assistance publique - Hôpitaux de Paris (AP-HP)-Association française contre les myopathies ( AFM-Téléthon ) -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 ) -Commissariat à l'énergie atomique et aux énergies alternatives ( CEA ) -Assistance publique - Hôpitaux de Paris (AP-HP)-Association française contre les myopathies ( AFM-Téléthon ) -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 Louis Pasteur - Strasbourg I-IFR37-Institut National de la Santé et de la Recherche Médicale ( INSERM ), and Assistance publique - Hôpitaux de Paris (AP-HP)-CHU Pitié-Salpêtrière [APHP]

Subjects
Male, Neuromyotonia, MESH: Mice, Mutant Strains, MESH: Muscle Contraction, Gene Dosage, medicine.disease_cause, MESH: Gene Dosage, MESH: Motor Endplate, chemistry.chemical_compound, Mice, 0302 clinical medicine, Missense mutation, MESH : Female, MESH : Gene Dosage, MESH: Animals, Genetics (clinical), 0303 health sciences, Mutation, biology, General Medicine, Acetylcholinesterase, MESH : Mice, Transgenic, MESH : Phenotype, medicine.anatomical_structure, Phenotype, MESH : Electromyography, Female, [SDV.NEU]Life Sciences [q-bio]/Neurons and Cognition [q-bio.NC], MESH : Heparan Sulfate Proteoglycans, Muscle Contraction, medicine.medical_specialty, MESH : Isaacs Syndrome, MESH: Mice, Transgenic, MESH : Male, Schwartz–Jampel syndrome, Mutation, Missense, Mice, Transgenic, MESH : Mice, Inbred C57BL, Perlecan, Osteochondrodysplasias, MESH: Phenotype, Motor Endplate, Neuromuscular junction, MESH : Acetylcholinesterase, MESH: Electromyography, 03 medical and health sciences, In vivo, MESH: Mice, Inbred C57BL, Internal medicine, MESH : Mice, Genetics, medicine, Animals, Humans, MESH: Isaacs Syndrome, Molecular Biology, MESH: Mice, Alleles, 030304 developmental biology, MESH : Osteochondrodysplasias, MESH: Mutation, Missense, MESH: Humans, Electromyography, MESH: Alleles, MESH : Humans, MESH : Motor Endplate, MESH: Acetylcholinesterase, medicine.disease, MESH: Osteochondrodysplasias, MESH : Disease Models, Animal, Mice, Mutant Strains, MESH: Male, MESH : Mice, Mutant Strains, Mice, Inbred C57BL, Disease Models, Animal, stomatognathic diseases, Endocrinology, chemistry, [ SDV.NEU ] Life Sciences [q-bio]/Neurons and Cognition [q-bio.NC], biology.protein, MESH: Heparan Sulfate Proteoglycans, MESH : Muscle Contraction, MESH : Animals, Isaacs Syndrome, MESH: Disease Models, Animal, MESH : Alleles, MESH: Female, MESH : Mutation, Missense, 030217 neurology & neurosurgery, Heparan Sulfate Proteoglycans
Abstract

International audience; Schwartz-Jampel syndrome (SJS) is a recessive neuromyotonia with chondrodysplasia. It results from hypomorphic mutations of the gene encoding perlecan, leading to a decrease in the levels of this heparan sulphate proteoglycan in basement membranes (BMs). It has been suggested that SJS neuromyotonia may result from endplate acetylcholinesterase (AChE) deficiency, but this hypothesis has never been investigated in vivo due to the lack of an animal model for neuromyotonia. We used homologous recombination to generate a knock-in mouse strain with one missense substitution, corresponding to a human familial SJS mutation (p.C1532Y), in the perlecan gene. We derived two lines, one with the p.C1532Y substitution alone and one with p.C1532Y and the selectable marker Neo, to down-regulate perlecan gene activity and to test for a dosage effect of perlecan in mammals. These two lines mimicked SJS neuromyotonia with spontaneous activity on electromyogramm (EMG). An inverse correlation between disease severity and perlecan secretion in the BMs was observed at the macroscopic and microscopic levels, consistent with a dosage effect. Endplate AChE levels were low in both lines, due to synaptic perlecan deficiency rather than major myofibre or neuromuscular junction disorganization. Studies of muscle contractile properties showed muscle fatigability at low frequencies of nerve stimulation and suggested that partial endplate AChE deficiency might contribute to SJS muscle stiffness by potentiating muscle force. However, physiological endplate AChE deficiency was not associated with spontaneous activity at rest on EMG in the diaphragm, suggesting that additional changes are required to generate such activity characteristic of SJS.

Published
2008
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5. Sostdc1 is expressed in all major compartments of developing and adult mammalian eyes

Author

Gabrielle Goldman, Marc Putterman, Fabien Guimiot, Thierry Jo Molina, Alexandre Moulin, Claudine Versaux-Botteri, Loïc Van Den Berghe, Francine Behar-Cohen, Alejandra Daruich, Eric Krejci, Matthieu P. Robert, Benoit Terris, Dominique Marchant, Christophe Klein, Maud Valensi, Laurent Jonet, Frederic Mascarelli, Dominique Bremond-Gignac, Marc Abitbol, EA no 2502 du ministère de la Recherche, de l'Enseignement Supérieur et la Technologie, Université Paris Descartes - Paris 5 (UPD5)-CERTO, Hormones, facteurs de croissance et physiopathologie vasculaire, Université Toulouse III - Paul Sabatier (UT3), Université Fédérale Toulouse Midi-Pyrénées-Université Fédérale Toulouse Midi-Pyrénées-IFR 31 Louis Bugnard (IFR 31), Centre National de la Recherche Scientifique (CNRS)-Institut National de la Santé et de la Recherche Médicale (INSERM)-CHU Toulouse [Toulouse]-Université Toulouse III - Paul Sabatier (UT3), Université Fédérale Toulouse Midi-Pyrénées-Université Fédérale Toulouse Midi-Pyrénées-Centre National de la Recherche Scientifique (CNRS)-Institut National de la Santé et de la Recherche Médicale (INSERM)-CHU Toulouse [Toulouse]-Institut National de la Santé et de la Recherche Médicale (INSERM), Développement, vieillissement et pathologie de la rétine, Institut National de la Santé et de la Recherche Médicale (INSERM), Department of Ophthalmology, Université de Lausanne (UNIL), Biologie des Jonctions Neuromusculaires Normales et Pathologiques (U686), 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 de Recherche des Cordeliers (CRC), Université Paris Diderot - Paris 7 (UPD7)-École pratique des hautes études (EPHE)-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 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), MINES ParisTech - École nationale supérieure des mines de Paris, Hôpital Robert Debré, Département de Pathologie, Assistance publique - Hôpitaux de Paris (AP-HP) (APHP)-Hôtel-Dieu-Université Paris Descartes - Paris 5 (UPD5), Université Sorbonne Paris Cité (USPC), CHU Necker - Enfants Malades [AP-HP], Centre de Recherche des Cordeliers (CRC (UMR_S_1138 / U1138)), École pratique des hautes études (EPHE)-Université Paris Diderot - Paris 7 (UPD7)-Université Paris Descartes - Paris 5 (UPD5)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Sorbonne Université (SU), IFR 31 Louis Bugnard (IFR 31), Université Fédérale Toulouse Midi-Pyrénées-Université Fédérale Toulouse Midi-Pyrénées-Centre National de la Recherche Scientifique (CNRS)-Institut National de la Santé et de la Recherche Médicale (INSERM)-CHU Toulouse [Toulouse]-Université Toulouse III - Paul Sabatier (UT3), Université Fédérale Toulouse Midi-Pyrénées-Université Fédérale Toulouse Midi-Pyrénées-Institut National de la Santé et de la Recherche Médicale (INSERM), Université Paris Diderot - Paris 7 (UPD7)-École pratique des hautes études (EPHE), Université Paris sciences et lettres (PSL)-Université Paris sciences et lettres (PSL)-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), Université Paris sciences et lettres (PSL), Assistance publique - Hôpitaux de Paris (AP-HP) (AP-HP)-Hôtel-Dieu-Université Paris Descartes - Paris 5 (UPD5), Assistance publique - Hôpitaux de Paris (AP-HP) (AP-HP), École pratique des hautes études (EPHE), and Université Paris sciences et lettres (PSL)-Université Paris sciences et lettres (PSL)-Université Paris Diderot - Paris 7 (UPD7)-Université Paris Descartes - Paris 5 (UPD5)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Sorbonne Université (SU)

Subjects
Male, Pathology, medicine.medical_specialty, genetic structures, [SDV]Life Sciences [q-bio], Blotting, Western, Retinal Pigment Epithelium, In situ hybridization, Biology, 03 medical and health sciences, Cellular and Molecular Neuroscience, SOX2, medicine, Animals, Humans, In Situ Hybridization, ComputingMilieux_MISCELLANEOUS, Adaptor Proteins, Signal Transducing, Aged, 030304 developmental biology, 0303 health sciences, Retina, Retinal pigment epithelium, 030305 genetics & heredity, Wnt signaling pathway, Gene Expression Regulation, Developmental, Eye Diseases, Hereditary, Immunohistochemistry, eye diseases, Sensory Systems, Rats, 3. Good health, Disease Models, Animal, Ophthalmology, medicine.anatomical_structure, Child, Preschool, Eye development, RNA, Female, sense organs, PAX6, Immunostaining
Abstract

This study was conducted in order to study Sostdc1 expression in rat and human developing and adult eyes.Using the yeast signal sequence trap screening method, we identified the Sostdc1 cDNA encoding a protein secreted by the adult rat retinal pigment epithelium. We determined by in situ hybridization, RT-PCR, immunohistochemistry, and western blot analysis Sostdc1 gene and protein expression in developing and postnatal rat ocular tissue sections. We also investigated Sostdc1 immunohistolocalization in developing and adult human ocular tissues.We demonstrated a prominent Sostdc1 gene expression in the developing rat central nervous system (CNS) and eyes at early developmental stages from E10.5 days postconception (dpc) to E13 dpc. Specific Sostdc1 immunostaining was also detected in most adult cells of rat ocular tissue sections. We also identified the rat ocular embryonic compartments characterized by a specific Sostdc1 immunohistostaining and specific Pax6, Sox2, Otx2, and Vsx2 immunohistostaining from embryonic stages E10.5 to E13 dpc. Furthermore, we determined the localization of SOSTDC1 immunoreactivity in ocular tissue sections of developing and adult human eyes. Indeed, we detected SOSTDC1 immunostaining in developing and adult human retinal pigment epithelium (RPE) and neural retina (NR) as well as in several developing and adult human ocular compartments, including the walls of choroidal and scleral vessels. Of utmost importance, we observed a strong SOSTDC1 expression in a pathological ocular specimen of type 2 Peters' anomaly complicated by retinal neovascularization as well in the walls ofother pathological extra-ocular vessels. CONCLUSION: As rat Sostdc1 and human SOSTDC1 are dual antagonists of the Wnt/β-catenin and BMP signaling pathways, these results underscore the potential crucial roles of these pathways and their antagonists, such as Sostdc1 and SOSTDC1, in developing and adult mammalian normal eyes as well as in syndromic and nonsyndromic congenital eye diseases.

Published
2019
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6. Smooth 2D manifold extraction from 3D image stack

Author

Asm Shihavuddin, Sreetama Basu, Elton Rexhepaj, Felipe Delestro, Nikita Menezes, Séverine M Sigoillot, Elaine Del Nery, Fekrije Selimi, Nathalie Spassky, Auguste Genovesio, Institut de biologie de l'ENS Paris (UMR 8197/1024) (IBENS), Département de Biologie - ENS Paris, École normale supérieure - Paris (ENS Paris)-École normale supérieure - Paris (ENS Paris)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Centre National de la Recherche Scientifique (CNRS), National University of Singapore (NUS), Biologie des Jonctions Neuromusculaires Normales et Pathologiques (U686), Université Paris Descartes - Paris 5 (UPD5)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Centre National de la Recherche Scientifique (CNRS), Translational Research Department, Institut Curie, PSL Research University, Paris 75248, France, Centre interdisciplinaire de recherche en biologie (CIRB), Centre National de la Recherche Scientifique (CNRS)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Collège de France (CdF)-PSL Research University (PSL), 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), 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)-Centre National de la Recherche Scientifique (CNRS), Labex MemoLife, 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 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), Institut de biologie de l'ENS Paris (IBENS), Centre National de la Recherche Scientifique (CNRS)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Département de Biologie - ENS Paris, and 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)

Subjects
[INFO.INFO-TS]Computer Science [cs]/Signal and Image Processing, Science, Article, ComputingMilieux_MISCELLANEOUS
Abstract

Three-dimensional fluorescence microscopy followed by image processing is routinely used to study biological objects at various scales such as cells and tissue. However, maximum intensity projection, the most broadly used rendering tool, extracts a discontinuous layer of voxels, obliviously creating important artifacts and possibly misleading interpretation. Here we propose smooth manifold extraction, an algorithm that produces a continuous focused 2D extraction from a 3D volume, hence preserving local spatial relationships. We demonstrate the usefulness of our approach by applying it to various biological applications using confocal and wide-field microscopy 3D image stacks. We provide a parameter-free ImageJ/Fiji plugin that allows 2D visualization and interpretation of 3D image stacks with maximum accuracy., Maximum Intensity Projection is a common tool to represent 3D biological imaging data in a 2D space, but it creates artefacts. Here the authors develop Smooth Manifold Extraction, an ImageJ/Fiji plugin, to preserve local spatial relationships when extracting the content of a 3D volume to a 2D space.

Published
2017
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7. Distribution of Smoothened at hippocampal mossy fiber synapses

Author

Martial Ruat, Bernard, Christelle Masdeu, Hélène Faure, Elisabeth Traiffort, Institut de Neurobiologie Alfred Fessard (INAF), Centre National de la Recherche Scientifique (CNRS), Laboratoire de neurobiologie cellulaire et moléculaire (NBCM), Biologie des Jonctions Neuromusculaires Normales et Pathologiques (U686), 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 - Faculté de Médecine (UPD5 Médecine), Université Paris Descartes - Paris 5 (UPD5), 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
Mossy fiber (hippocampus), Patched, G-protein-coupled receptor, hedgehog, [SDV.NEU.NB]Life Sciences [q-bio]/Neurons and Cognition [q-bio.NC]/Neurobiology, MESH: Membrane Glycoproteins, Nerve Tissue Proteins, MESH: Receptors, G-Protein-Coupled, Biology, Hippocampal formation, Receptors, G-Protein-Coupled, Mice, 03 medical and health sciences, 0302 clinical medicine, Microscopy, Electron, Transmission, synaptic vesicles, medicine, Animals, MESH: Animals, [SDV.NEU] Life Sciences [q-bio]/Neurons and Cognition [q-bio.NC], dentate gyrus, MESH: Nerve Tissue Proteins, patched, MESH: Mice, 030304 developmental biology, Hippocampal mossy fiber, 0303 health sciences, Membrane Glycoproteins, General Neuroscience, Dentate gyrus, [SDV.NEU.NB] Life Sciences [q-bio]/Neurons and Cognition [q-bio.NC]/Neurobiology, MESH: Mossy Fibers, Hippocampal, Granule cell, Smoothened Receptor, bouton, medicine.anatomical_structure, MESH: Subcellular Fractions, Mossy Fibers, Hippocampal, MESH: Microscopy, Electron, Transmission, [SDV.NEU]Life Sciences [q-bio]/Neurons and Cognition [q-bio.NC], Smoothened, Neuroscience, 030217 neurology & neurosurgery, Subcellular Fractions, MESH: Dentate Gyrus
Abstract

The seven-transmembrane receptor Smoothened is essential for hedgehog signal transduction. In adulthood, the highest density of Smoothened mRNA is found in the granule cell layer of the dentate gyrus. There, Smoothened expression is regulated by the synaptic activity involving the glutamatergic transmission. The precise localization of Smoothened proteins, however, has not yet been reported. Here, we describe Smoothened protein distribution in the hippocampal mossy fibers using specific Smoothened antibodies. We provide evidences for their presynaptic localization, and using electron microscopy, show that Smoothened is located in close association with synaptic vesicles and rarely with the plasma membrane. These findings demonstrate that Smoothened is localized presynaptically and suggest that Smoothened signal transduction may be implicated in the complex aspects of mossy fiber function.

Published
2007
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8. Wnt4 participates in the formation of vertebrate neuromuscular junction

Author

Claire Legay, Jérôme Rouvière, Laure Strochlic, Francine Bourgeois, Laurent Schaeffer, Séverine M. Sigoillot, Amanda Swain, Valérie Castellani, Perrine Delers, Julien Falk, Evelyne Goillot, Institut Jacques Monod (IJM (UMR_7592)), Université Paris Diderot - Paris 7 (UPD7)-Centre National de la Recherche Scientifique (CNRS), Centre de génétique et de physiologie moléculaire et cellulaire (CGPhiMC), Centre National de la Recherche Scientifique (CNRS)-Université Claude Bernard Lyon 1 (UCBL), Université de Lyon-Université de Lyon, Laboratoire de Biologie Moléculaire de la Cellule (LBMC), Institut National de la Santé et de la Recherche Médicale (INSERM)-Centre National de la Recherche Scientifique (CNRS)-École normale supérieure - Lyon (ENS Lyon)-Université Claude Bernard Lyon 1 (UCBL), Biologie des Jonctions Neuromusculaires Normales et Pathologiques (U686), Université Paris Descartes - Paris 5 (UPD5)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Centre National de la Recherche Scientifique (CNRS), Laviron, Nathalie, Université Claude Bernard Lyon 1 (UCBL), Université de Lyon-Université de Lyon-Centre National de la Recherche Scientifique (CNRS), 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), Institut Jacques Monod (IJM), Université Paris Diderot - Paris 7 (UPD7) - Centre National de la Recherche Scientifique (CNRS), Université de Lyon - Université de Lyon - Centre National de la Recherche Scientifique (CNRS), 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), 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 École normale supérieure de Lyon (ENS de Lyon)-Université Claude Bernard Lyon 1 (UCBL)

Subjects
MESH: Muscles, MESH : Vertebrates, Synaptogenesis, Biochemistry, MESH : Phosphorylation, Mice, 0302 clinical medicine, WNT4, Receptors, Cholinergic, MESH: Animals, MESH: Vertebrates, Cholinergic, 0303 health sciences, MESH : Protein Binding, Cell biology, MESH: COS Cells, MESH: HEK293 Cells, COS Cells, Medicine, Science, Neuromuscular junction, 03 medical and health sciences, MESH : Cluster Analysis, Humans, MESH: Protein Binding, Biology, Acetylcholine receptor, MESH : Muscle Cells, MESH: Receptors, Cholinergic, MESH: Humans, MESH: Phosphorylation, MESH : Humans, Proteins, Receptor Protein-Tyrosine Kinases, [SDV.BBM.BM]Life Sciences [q-bio]/Biochemistry, Molecular Biology/Molecular biology, Molecular Development, MESH : Receptors, Cholinergic, MESH: Cercopithecus aethiops, MESH: Cluster Analysis, MESH : HEK293 Cells, MESH : Muscles, Immunology, MESH : Neuromuscular Junction, MESH : Cercopithecus aethiops, MESH : Gene Expression Regulation, Developmental, MESH: Neuromuscular Junction, Animals, Biological Markers, Body Patterning, Cercopithecus aethiops, Cluster Analysis, Embryo, Mammalian, Gene Expression Regulation, Developmental, HEK293 Cells, Muscle Cells, Muscles, Neuromuscular Junction, Phosphorylation, Protein Binding, Receptors, Vertebrates, Wnt4 Protein, 030217 neurology & neurosurgery, Biomarkers, Developmental Biology, Neuroscience, MESH: Muscle Cells, [SDV.BBM.BM] Life Sciences [q-bio]/Biochemistry, Molecular Biology/Molecular biology, Postsynaptic potential, Molecular Cell Biology, Chlorocebus aethiops, MESH: Gene Expression Regulation, Developmental, MESH : Wnt4 Protein, Myocyte, MESH : Embryo, Mammalian, Multidisciplinary, Muscle cell differentiation, Myogenesis, Gene Expression Regulation, Developmental, Signaling Cascades, medicine.anatomical_structure, MESH: Receptor Protein-Tyrosine Kinases, SDV:BBM:BM, MESH : COS Cells, Research Article, Signal Transduction, MESH: Body Patterning, animal structures, Developmental Neuroscience, MESH : Mice, medicine, MESH: Wnt4 Protein, MESH: Mice, 030304 developmental biology, MESH: Biological Markers, MESH: Embryo, Mammalian, MESH : Body Patterning, MESH : Receptor Protein-Tyrosine Kinases, Embryo, Mammalian, MESH : Biological Markers, MESH : Animals
Abstract

International audience; Neuromuscular junction (NMJ) formation requires the highly coordinated communication of several reciprocal signaling processes between motoneurons and their muscle targets. Identification of the early, spatially restricted cues in target recognition at the NMJ is still poorly documented, especially in mammals. Wnt signaling is one of the key pathways regulating synaptic connectivity. Here, we report that Wnt4 contributes to the formation of vertebrate NMJ in vivo. Results from a microarray screen and quantitative RT-PCR demonstrate that Wnt4 expression is regulated during muscle cell differentiation in vitro and muscle development in vivo, being highly expressed when the first synaptic contacts are formed and subsequently downregulated. Analysis of the mouse Wnt4⁻/⁻ NMJ phenotype reveals profound innervation defects including motor axons overgrowing and bypassing AChR aggregates with 30% of AChR clusters being unapposed by nerve terminals. In addition, loss of Wnt4 function results in a 35% decrease of the number of prepatterned AChR clusters while Wnt4 overexpression in cultured myotubes increases the number of AChR clusters demonstrating that Wnt4 directly affects postsynaptic differentiation. In contrast, muscle structure and the localization of several synaptic proteins including acetylcholinesterase, MuSK and rapsyn are not perturbed in the Wnt4 mutant. Finally, we identify MuSK as a Wnt4 receptor. Wnt4 not only interacts with MuSK ectodomain but also mediates MuSK activation. Taken together our data reveal a new role for Wnt4 in mammalian NMJ formation that could be mediated by MuSK, a key receptor in synaptogenesis.

Published
2012
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9. Role of the specific collagen ColQ in the neuromuscular junction formation

Author

Sigoillot, Séverine, Sigoillot, Séverine, Biologie des Jonctions Neuromusculaires Normales et Pathologiques (U686), 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 VI, Claire Legay(claire.legay@parisdescartes.fr), 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
Nicotinic acetylcholine receptor, Syndrome myasthénique congénital, Cholinestérases, Jonction neuromusculaire, Neuromuscular junction, ColQ, [SDV.BC]Life Sciences [q-bio]/Cellular Biology, Congenital myasthenic syndrome, [SDV.BBM.BC]Life Sciences [q-bio]/Biochemistry, Molecular Biology/Biomolecules [q-bio.BM], [SDV.BBM.BC] Life Sciences [q-bio]/Biochemistry, Molecular Biology/Biomolecules [q-bio.BM], Récepteur nicotinique de l'acétylcholine, [SDV.NEU]Life Sciences [q-bio]/Neurons and Cognition [q-bio.NC], [SDV.NEU] Life Sciences [q-bio]/Neurons and Cognition [q-bio.NC], [SDV.BBM.BC] Life Sciences [q-bio]/Biochemistry, Molecular Biology/Biochemistry [q-bio.BM], [SDV.BC] Life Sciences [q-bio]/Cellular Biology, MuSK
Abstract

ColQ is a specific collagen that anchors acetylcholinesterase (AChE) in the synaptic cleft of the neuromuscular junction. The importance of AChE-ColQ complex in the physiology of this synapse has been highlighted by the identification of COLQ mutations in the human gene, leading to a congenital myasthenic syndrome with AChE deficiency (CMS-1c). The symptoms observed in patients and the defects of the neuromuscular junctions in adult ColQ mutant mice which are mice model for this CMS-1c are complex and AChE deficiency cannot account for all of them. We hypothesized that ColQ could play a role per se in synapse formation and therefore its absence would participate in the defects. I have shown that ColQ regulates the levels of nicotinic acetylcholine receptor (nAChR) subunits via MuSK, a key protein in synapse formation, independently of AChE. The consequence of this regulation is that ColQ controls nAChR clustering implicated in synaptic transmission. Moreover, my results have shown that ColQ deficiency reduces membrane-bound MuSK. In this context, it is highly significant that the phenotypes of CMS-1c and MuSK CMS patients have similarities. In addition, atrophic signs were found in ColQ-/- mice muscles suggesting a partial denervation of muscle fibers in the absence of ColQ. Modifications highlighted by my results may thus explain the origin of the functional defects linked to ColQ absence or mutation., ColQ est un collagène spécifique ancrant l'acétylcholinestérase (AChE) dans la fente synaptique de la jonction neuromusculaire (JNM). L'importance du complexe AChE-ColQ dans le fonctionnement de cette synapse a été révélée par l'identification de mutations dans le gène humain codant pour ColQ qui sont à l'origine du syndrome myasthénique congénital avec déficience en AChE (SMC-1c). Les défauts présents chez les patients et le phénotype des souris ColQ-/- modèles pour le SMC-1c sont complexes, la déficience en AChE ne rendant probablement pas compte de tous les symptômes. Nous avons donc fait l'hypothèse que ColQ pourrait réguler la formation de la JNM afin d'expliquer certains des défauts observés. J'ai montré que ColQ régule l'expression des sous-unités du récepteur de l'acétylcholine (RACh) via le récepteur tyrosine kinase MuSK, molécule clé dans la formation de la synapse et cela indépendamment de l'AChE. La conséquence de cette régulation est que ColQ contrôle la formation des agrégats de RACh impliqués dans la transmission synaptique. Par ailleurs, mes résultats ont révélé que l'absence de ColQ entraîne une réduction de l'expression de MuSK à la membrane. Cette observation prend tout son intérêt dans un contexte clinique montrant que certains symptômes sont communs aux patients atteints du SMC-1c et du SMC lié à des mutations de MuSK. De plus, des signes d'atrophie musculaire ont été découverts chez la souris ColQ-/- pouvant laisser penser à une dénervation au moins partielle du muscle en absence de ColQ. Mes recherches ont ainsi permis de mettre en évidence des modifications pouvant être à l'origine des défauts fonctionnels observés lorsque ColQ est muté ou absent.

Published
2010

10. Rôle du collagène spécifique ColQ dans la formation de la jonction neuromusculaire

Author

Sigoillot, Séverine, Sigoillot, Séverine, Biologie des Jonctions Neuromusculaires Normales et Pathologiques (U686), 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 VI, and Claire Legay(claire.legay@parisdescartes.fr)

Subjects
Nicotinic acetylcholine receptor, Syndrome myasthénique congénital, Cholinestérases, Jonction neuromusculaire, Neuromuscular junction, ColQ, [SDV.BC]Life Sciences [q-bio]/Cellular Biology, Congenital myasthenic syndrome, Récepteur nicotinique de l'acétylcholine, [SDV.NEU]Life Sciences [q-bio]/Neurons and Cognition [q-bio.NC], [SDV.NEU] Life Sciences [q-bio]/Neurons and Cognition [q-bio.NC], [SDV.BBM.BC]Life Sciences [q-bio]/Biochemistry, Molecular Biology/Biochemistry [q-bio.BM], [SDV.BBM.BC] Life Sciences [q-bio]/Biochemistry, Molecular Biology/Biochemistry [q-bio.BM], [SDV.BC] Life Sciences [q-bio]/Cellular Biology, MuSK
Abstract

ColQ is a specific collagen that anchors acetylcholinesterase (AChE) in the synaptic cleft of the neuromuscular junction. The importance of AChE-ColQ complex in the physiology of this synapse has been highlighted by the identification of COLQ mutations in the human gene, leading to a congenital myasthenic syndrome with AChE deficiency (CMS-1c). The symptoms observed in patients and the defects of the neuromuscular junctions in adult ColQ mutant mice which are mice model for this CMS-1c are complex and AChE deficiency cannot account for all of them. We hypothesized that ColQ could play a role per se in synapse formation and therefore its absence would participate in the defects. I have shown that ColQ regulates the levels of nicotinic acetylcholine receptor (nAChR) subunits via MuSK, a key protein in synapse formation, independently of AChE. The consequence of this regulation is that ColQ controls nAChR clustering implicated in synaptic transmission. Moreover, my results have shown that ColQ deficiency reduces membrane-bound MuSK. In this context, it is highly significant that the phenotypes of CMS-1c and MuSK CMS patients have similarities. In addition, atrophic signs were found in ColQ-/- mice muscles suggesting a partial denervation of muscle fibers in the absence of ColQ. Modifications highlighted by my results may thus explain the origin of the functional defects linked to ColQ absence or mutation., ColQ est un collagène spécifique ancrant l'acétylcholinestérase (AChE) dans la fente synaptique de la jonction neuromusculaire (JNM). L'importance du complexe AChE-ColQ dans le fonctionnement de cette synapse a été révélée par l'identification de mutations dans le gène humain codant pour ColQ qui sont à l'origine du syndrome myasthénique congénital avec déficience en AChE (SMC-1c). Les défauts présents chez les patients et le phénotype des souris ColQ-/- modèles pour le SMC-1c sont complexes, la déficience en AChE ne rendant probablement pas compte de tous les symptômes. Nous avons donc fait l'hypothèse que ColQ pourrait réguler la formation de la JNM afin d'expliquer certains des défauts observés. J'ai montré que ColQ régule l'expression des sous-unités du récepteur de l'acétylcholine (RACh) via le récepteur tyrosine kinase MuSK, molécule clé dans la formation de la synapse et cela indépendamment de l'AChE. La conséquence de cette régulation est que ColQ contrôle la formation des agrégats de RACh impliqués dans la transmission synaptique. Par ailleurs, mes résultats ont révélé que l'absence de ColQ entraîne une réduction de l'expression de MuSK à la membrane. Cette observation prend tout son intérêt dans un contexte clinique montrant que certains symptômes sont communs aux patients atteints du SMC-1c et du SMC lié à des mutations de MuSK. De plus, des signes d'atrophie musculaire ont été découverts chez la souris ColQ-/- pouvant laisser penser à une dénervation au moins partielle du muscle en absence de ColQ. Mes recherches ont ainsi permis de mettre en évidence des modifications pouvant être à l'origine des défauts fonctionnels observés lorsque ColQ est muté ou absent.

Published
2010

11. Targeting of Acetylcholinesterase in Neurons In Vivo: A Dual Processing Function for the Proline-Rich Membrane Anchor Subunit and the Attachment Domain on the Catalytic Subunit

Author

Eric Krejci, Shelley Camp, Korami Dembele, Anna Hrabovska, Alexandre Dobbertin, Palmer Taylor, Véronique Bernard, Institut de Physique et Chimie des Matériaux de Strasbourg (IPCMS), Université de Strasbourg (UNISTRA)-Matériaux et nanosciences d'Alsace (FMNGE), Institut de Chimie du CNRS (INC)-Université de Strasbourg (UNISTRA)-Université de Haute-Alsace (UHA) Mulhouse - Colmar (Université de Haute-Alsace (UHA))-Institut National de la Santé et de la Recherche Médicale (INSERM)-Centre National de la Recherche Scientifique (CNRS)-Institut de Chimie du CNRS (INC)-Université de Strasbourg (UNISTRA)-Université de Haute-Alsace (UHA) Mulhouse - Colmar (Université de Haute-Alsace (UHA))-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)-Réseau nanophotonique et optique, Centre National de la Recherche Scientifique (CNRS)-Université de Strasbourg (UNISTRA)-Université de Haute-Alsace (UHA) Mulhouse - Colmar (Université de Haute-Alsace (UHA))-Centre National de la Recherche Scientifique (CNRS)-Université de Strasbourg (UNISTRA), Biologie des Jonctions Neuromusculaires Normales et Pathologiques (U686), Université Paris Descartes - Paris 5 (UPD5)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Centre National de la Recherche Scientifique (CNRS), Neurosciences Paris Seine (NPS), Sorbonne Université (SU)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Institut de Biologie Paris Seine (IBPS), and Sorbonne Université (SU)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Centre National de la Recherche Scientifique (CNRS)-Sorbonne Université (SU)-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)

Subjects
Male, Protein subunit, Molecular Sequence Data, Nerve Tissue Proteins, Biology, Article, Cell Line, 03 medical and health sciences, chemistry.chemical_compound, Mice, 0302 clinical medicine, Catalytic Domain, Enzyme Stability, Animals, Humans, Amino Acid Sequence, Binding site, ComputingMilieux_MISCELLANEOUS, 030304 developmental biology, Mice, Knockout, Neurons, 0303 health sciences, Binding Sites, Base Sequence, General Neuroscience, Endoplasmic reticulum, Wild type, Membrane Proteins, Acetylcholinesterase, Transmembrane protein, humanities, Cell biology, Membrane protein, chemistry, Biochemistry, Gene Targeting, Female, [SDV.NEU]Life Sciences [q-bio]/Neurons and Cognition [q-bio.NC], 030217 neurology & neurosurgery, Intracellular
Abstract

Acetylcholinesterase (AChE) accumulates on axonal varicosities and is primarily found as tetramers associated with a proline-rich membrane anchor (PRiMA). PRiMA is a small transmembrane protein that efficiently transforms secreted AChE to an enzyme anchored on the outer cell surface. Surprisingly, in the striatum of the PRiMA knock-out mouse, despite a normal level of AChE mRNA, we find only 2–3% of wild type AChE activity, with the residual AChE localized in the endoplasmic reticulum, demonstrating that PRiMAin vivois necessary for intracellular processing of AChE in neurons. Moreover, deletion of the retention signal of the AChE catalytic subunit in mice, which is the domain of interaction with PRiMA, does not restore AChE activity in the striatum, establishing that PRiMA is necessary to target and/or to stabilize nascent AChE in neurons. These unexpected findings open new avenues to modulating AChE activity and its distribution in CNS disorders.

Published
2009
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12. Influence of differential expression of acetylcholinesterase in brain and muscle on respiration

Author

Arthur S. Foutz, Palmer Taylor, Shelley Camp, Jean Champagnat, Eliane Boudinot, Véronique Bernard, Eric Krejci, Neurobiologie génétique et intégrative (NGI), Centre National de la Recherche Scientifique (CNRS), Institut de Neurobiologie Alfred Fessard (INAF), Biologie des Jonctions Neuromusculaires Normales et Pathologiques (U686), Université Paris Descartes - Paris 5 (UPD5)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Centre National de la Recherche Scientifique (CNRS), Department of pharmacology, University of California [San Diego] (UC San Diego), University of California-University of California, 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
Male, MESH: Muscles, MESH: Body Temperature, Physiology, MESH: Anoxia, MESH: Pulmonary Ventilation, MESH: Tidal Volume, MESH: Mice, Knockout, Body Temperature, Hypercapnia, Mice, chemistry.chemical_compound, 0302 clinical medicine, Respiratory function, MESH: Animals, Respiratory system, Hypoxia, Butyrylcholinesterase, MESH: Plethysmography, Whole Body, Sequence Deletion, Mice, Knockout, 0303 health sciences, MESH: Butyrylcholinesterase, Muscles, Respiration, General Neuroscience, Brain, Exons, MESH: Sequence Deletion, Acetylcholinesterase, MESH: Gene Expression Regulation, Bronchodilator Agents, medicine.anatomical_structure, Knockout mouse, Female, [SDV.NEU]Life Sciences [q-bio]/Neurons and Cognition [q-bio.NC], MESH: Terbutaline, Acetylcholine, medicine.drug, Pulmonary and Respiratory Medicine, medicine.medical_specialty, Biology, Article, Neuromuscular junction, 03 medical and health sciences, MESH: Brain, Internal medicine, MESH: Analysis of Variance, Terbutaline, Tidal Volume, medicine, Animals, [SDV.NEU] Life Sciences [q-bio]/Neurons and Cognition [q-bio.NC], MESH: Mice, Gene knockout, Plethysmography, Whole Body, 030304 developmental biology, MESH: Respiration, Analysis of Variance, MESH: Acetylcholinesterase, MESH: Male, Endocrinology, Gene Expression Regulation, chemistry, MESH: Hypercapnia, MESH: Bronchodilator Agents, Pulmonary Ventilation, MESH: Exons, MESH: Female, 030217 neurology & neurosurgery
Abstract

International audience; A mouse strain with a deleted acetylcholinesterase (AChE) gene (AChE knockout) shows a decreased inspiration time and increased tidal volume and ventilation .To investigate the respective roles of AChE in brain and muscle, we recorded respiration by means of whole-body plethysmography in knockout mice with tissue selective deletions in AChE expression. A mouse strain with the anchoring domains of AChE deleted (del E5+6 knockout mice) has very low activity in the brain and neuromuscular junction, but increased monomeric AChE in serum. A mouse strain with deletion of the muscle specific region of AChE (del i1RR knockout mice) exhibits no expression in muscle, but unaltered expression in the central nervous system. Neither strain exhibits the pronounced phenotypic traits observed in the complete AChE knockout strain. A third strain lacking the anchor molecule PRiMA, has no functional AChE and butyrylcholinesterase (BChE) in brain and an unaltered respiratory function. BChE inhibition by bambuterol decreases tidal volume and body temperature in del E5+6 and i1RR knockout strains, but not in PRiMA deletion or wild-type controls. We find that: (1) deletion of the full AChE gene is required for a pronounced alteration in respiratory phenotype, (2) BChE is involved in respiratory muscles contraction and temperature control in del E5+6 and i1RR knockout mice, and (3) AChE expression requiring a gene product splice to either exons 5 and 6 or regulated by intron1 influences temperature control.

Published
2009
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13. La protéine Sonic Hedgehog: un nouveau modulateur de l'activité neuronale?

Author

Christelle Masdeu, Olivier Pascual, Jean Champagnat, Véronique Bernard, Hélène Faure, Martial Ruat, Elisabeth Traiffort, Institut de Neurobiologie Alfred Fessard (INAF), Centre National de la Recherche Scientifique (CNRS), Laboratoire de neurobiologie cellulaire et moléculaire (NBCM), Neurobiologie génétique et intégrative (NGI), Biologie des Jonctions Neuromusculaires Normales et Pathologiques (U686), 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 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
[SDV.NEU]Life Sciences [q-bio]/Neurons and Cognition [q-bio.NC], [SDV.NEU] Life Sciences [q-bio]/Neurons and Cognition [q-bio.NC]
Published
2007

14. Butyrylcholinesterase and the control of synaptic responses in acetylcholinesterase knockout mice

Author

Eric Krejci, Jordi Molgó, Emmanuelle Girard, Véronique Bernard, Jasmina Minic, Arnaud Chatonnet, Laboratoire de neurobiologie cellulaire et moléculaire (NBCM), Centre National de la Recherche Scientifique (CNRS), Institut de Neurobiologie Alfred Fessard (INAF), Biologie des Jonctions Neuromusculaires Normales et Pathologiques (U686), Université Paris Descartes - Paris 5 (UPD5)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Centre National de la Recherche Scientifique (CNRS), Département de Physiologie Animale, Institut National de la Recherche Agronomique (INRA), Université Paris Descartes - Paris 5 (UPD5) - Institut National de la Santé et de la Recherche Médicale (INSERM) - Centre National de la Recherche Scientifique (CNRS), Dynamique Musculaire et Métabolisme (DMEM), and Institut National de la Recherche Agronomique (INRA)-Université de Montpellier (UM)

Subjects
Male, MESH: Tetraisopropylpyrophosphamide, Time Factors, Nicotinic acetylcholine receptors, Neuromuscular junction, Synaptic Transmission, MESH: Mice, Knockout, MESH: Motor Endplate, Mice, chemistry.chemical_compound, Perisynaptic schwann cells, 0302 clinical medicine, MESH: Animals, General Pharmacology, Toxicology and Pharmaceutics, Butyrylcholinesterase, Mice, Knockout, 0303 health sciences, MESH: Muscle, Skeletal, MESH: Electrophysiology, MESH: Butyrylcholinesterase, Chemistry, General Medicine, Acetylcholinesterase, Cell biology, Electrophysiology, medicine.anatomical_structure, Biochemistry, Female, [SDV.NEU]Life Sciences [q-bio]/Neurons and Cognition [q-bio.NC], MESH: Cholinesterase Inhibitors, Acetylcholine, medicine.drug, Synaptic cleft, Mice, Inbred Strains, MESH: Microscopy, Electron, Neurotransmission, Motor Endplate, MESH: Mice, Inbred Strains, General Biochemistry, Genetics and Molecular Biology, 03 medical and health sciences, medicine, MESH: Synaptic Transmission, Animals, MESH: Benzenaminium, 4,4'-(3-oxo-1,5-pentanediyl)bis(N,N-dimethyl-N-2-propenyl-), Dibromide, [SDV.NEU] Life Sciences [q-bio]/Neurons and Cognition [q-bio.NC], Muscle, Skeletal, MESH: Mice, 030304 developmental biology, Acetylcholine receptor, Tetraisopropylpyrophosphamide, MESH: Time Factors, Cholinesterase inhibitors, Benzenaminium, 4,4'-(3-oxo-1,5-pentanediyl)bis(N,N-dimethyl-N-2-propenyl-), Dibromide, MESH: Acetylcholinesterase, MESH: Male, Microscopy, Electron, Acetylcholinesterase knockout mice, MESH: Neuromuscular Junction, MESH: Female, 030217 neurology & neurosurgery
Abstract

At the neuromuscular junction (NMJ) acetylcholinesterase (AChE) and butyrylcholinesterase (BChE) can hydrolyze acetylcholine (ACh). Released ACh quanta are known to diffuse rapidly across the narrow synaptic cleft and pairs of ACh molecules cooperate to open endplate channels. During their diffusion through the cleft, or after being released from muscle nicotinic ACh receptors (nAChRs), most ACh molecules are hydrolyzed by AChE highly concentrated at the NMJ. Advances in mouse genomics offered new approaches to assess the role of specific cholinesterases involved in synaptic transmission. AChE knockout mice (AChE-KO) provide a valuable tool for examining the complete abolition of AChE activity and the role of BChE. AChE-KO mice live to adulthood, and exhibit an increased sensitivity to BChE inhibitors, suggesting that BChE activity facilitated their survival and compensated for AChE function. Our results show that BChE is present at the endplate region of wild-type and AChE-KO mature muscles. The decay time constant of focally recorded miniature endplate currents was 1.04 +/- 0.06 ms in wild-type junctions and 5.4 ms +/- 0.3 ms in AChE-KO junctions, and remained unaffected by BChE-specific inhibitors, indicating that BChE is not limiting ACh duration on endplate nAChRs. Inhibition of BChE decreased evoked quantal ACh release in AChE-KO NMJs. This reduction in ACh release can explain the greatest sensitivity of AChE-KO mice to BChE inhibitors. BChE is known to be localized in perisynaptic Schwann cells, and our results strongly suggest that BChE's role at the NMJ is to protect nerve terminals from an excess of ACh.

Published
2007
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15. In vitro expression, kinetic, pharmacologic, molecular, and evolutionary analyses, and molecular modeling of a recombinant acetylcholinesterase from the invertebrate urochordate ciona intestinalis: assembly of asymetric forms with colq

Author

Frederick, A., Tsigelny, I., Cohenour, F., Spiker, C., Krejci, E., Chatonnet, Arnaud, Bourgoin, Sylvain, Richards, R., Allen, Thomas, Whitlock, M.H., Pezzementi, L., Department of Biology, Birmingham-Southern College, Department of Chemistry and Biochemistry, University of California [San Diego] (UC San Diego), University of California-University of California, Biologie des Jonctions Neuromusculaires Normales et Pathologiques (U686), Université Paris Descartes - Paris 5 (UPD5)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Centre National de la Recherche Scientifique (CNRS), Différenciation Cellulaire et Croissance (DCC), and Institut National de la Recherche Agronomique (INRA)-Université Montpellier 2 - Sciences et Techniques (UM2)

Subjects
[SDV]Life Sciences [q-bio], PHYSIOLOGIE HUMAINE, CHOLINESTERASE, PHYLOGENETIC ANALYSIS, [INFO]Computer Science [cs], COLLAGEN-LIKE TAIL SUBUNIT, COLQ, BIOLOGIE MOLECULAIRE, EVOLUTION
Abstract

International audience; To learn more about the evolution of the cholinesterases, acetylcholinesterase (AChE) and butyrylcholinesterase (BuChE) in the vertebrates, we investigated the AChE activity of a deuterostome invertebrate, the urochordate Ciona intestinalis, by expressing in vitro a synthetic recombinant cDNA for the enzyme in COS-7 cells. On the basis of evidence from kinetics, pharmacology, molecular biology and molecular modeling, we confirm that the enzyme is AChE. Sequence analysis also suggests that the cDNA is AChET and should be able to code for all three globular forms of AChE: monomers (G1), dimers (G2), and tetramers (G4); and, assemble into asymmetric forms in association with the collagenic subunit colQ. Using velocity sedimentation on sucrose gradients, we found that all three of the globular forms are either expressed in cells or secreted into the medium. In cells, amphiphilic G1a and non-amphiphilic G4na forms are found. Amphiphilic G2a and non-amphiphilic G4na forms are secreted into the medium. When the catalytic subunit is co-expressed with rat colQ the A12 form of the enzyme is expressed. Collagenase digestion of the A12 AChE at 37°C produces a lytic G4 form; only globular forms are present in vivo. This is the first demonstration that an invertebrate AChE is capable of assembling into asymmetric forms. We also performed a phylogenetic analysis of the sequence. We will discuss the relevance of our results to the evolution of the cholinesterases in general, in deuterostome invertebrates, and in chordates including vertebrates.

Published
2007

16. Intraneuronal trafficking of G-protein-coupled receptors in vivo

Author

Marion Decossas, Véronique Bernard, Isabel Liste, Bertrand Bloch, Biologie des Jonctions Neuromusculaires Normales et Pathologiques (U686), 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 d'Histologie-Embryologie (UMR 5541), Centre National de la Recherche Scientifique (CNRS), Immunologie et chimie thérapeutiques (ICT), Cancéropôle du Grand Est - Centre National de la Recherche Scientifique (CNRS), Center of Molecular Biology Severo Ochoa, University of Madrid, 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 Cancéropôle du Grand Est-Centre National de la Recherche Scientifique (CNRS)

Subjects
Intracellular Fluid, MESH: Signal Transduction, MESH: Protein Transport, G protein, [SDV.NEU.NB]Life Sciences [q-bio]/Neurons and Cognition [q-bio.NC]/Neurobiology, MESH: Neurons, Down-Regulation, MESH: Receptors, G-Protein-Coupled, Biology, MESH: Neurotransmitter Agents, MESH: Down-Regulation, Receptors, G-Protein-Coupled, 03 medical and health sciences, chemistry.chemical_compound, MESH: Brain, 0302 clinical medicine, Cell surface receptor, medicine, Animals, Humans, MESH: Animals, Axon, Receptor, Neurotransmitter, 030304 developmental biology, G protein-coupled receptor, Neurons, 0303 health sciences, Neurotransmitter Agents, MESH: Humans, Staining and Labeling, MESH: Intracellular Fluid, General Neuroscience, Cell Membrane, [SDV.NEU.NB] Life Sciences [q-bio]/Neurons and Cognition [q-bio.NC]/Neurobiology, Brain, Endocytosis, Protein Transport, MESH: Staining and Labeling, medicine.anatomical_structure, chemistry, nervous system, MESH: Endocytosis, Signal transduction, Neuroscience, 030217 neurology & neurosurgery, Signal Transduction, MESH: Cell Membrane
Abstract

In vitro studies have widely demonstrated that the abundance and availability of G-protein-coupled receptors (GPCRs) at the cell surface is regulated by the neuronal environment and is the result of complex intraneuronal trafficking. However, this regulation is still poorly understood in vivo. Modulation of receptor availability at the neuronal membrane is a key event in the regulation of neuronal functions (e.g. neurotransmitter release or neuronal excitability in physiological, pathological or therapeutic conditions). We discuss the effects of duration of receptor stimulation (acute versus chronic) on the intraneuronal trafficking of GPCRs in vivo, and we show that this trafficking might differ according to subcellular compartment (soma, dendrites or axon terminals).

Published
2006
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17. Generation of a mouse model for Schwartz-Jampel syndrome

Author

Stum, Morgane, Girard, Emmanuelle, Molgó, Jordi, Tabti, Nacira, Willer, Jean-Claude, Fontaine, Bertrand, Krejci, Eric, Nicole, Sophie, Outters-Lafaye, Michèle, Laboratoire de neurobiologie cellulaire et moléculaire (NBCM), Centre National de la Recherche Scientifique (CNRS), Institut de Neurobiologie Alfred Fessard (INAF), Biologie des Jonctions Neuromusculaires Normales et Pathologiques (U686), 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 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
[SDV.NEU]Life Sciences [q-bio]/Neurons and Cognition [q-bio.NC], [SDV.NEU] Life Sciences [q-bio]/Neurons and Cognition [q-bio.NC]
Published
2006

18. Remodeling of the neuromuscular junction in mice with deleted exons 5 and 6 of acetylcholinesterase

Author

Palmer Taylor, Véronique Bernard, Eric Krejci, Emmanuelle Girard, Shelley Camp, Jordi Molgó, Laboratoire de neurobiologie cellulaire et moléculaire (NBCM), Centre National de la Recherche Scientifique (CNRS), Institut de Neurobiologie Alfred Fessard (INAF), Biologie des Jonctions Neuromusculaires Normales et Pathologiques (U686), Université Paris Descartes - Paris 5 (UPD5)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Centre National de la Recherche Scientifique (CNRS), Department of pharmacology, University of California [San Diego] (UC San Diego), University of California-University of California, 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
[SDV.NEU.NB]Life Sciences [q-bio]/Neurons and Cognition [q-bio.NC]/Neurobiology, Diaphragm, MESH: Respiratory Muscles, Neuromuscular transmission, Neuromuscular Junction, Biology, Receptors, Nicotinic, MESH: Mice, Knockout, Neuromuscular junction, 03 medical and health sciences, Cellular and Molecular Neuroscience, chemistry.chemical_compound, Exon, Mice, 0302 clinical medicine, COLQ, medicine, Coding region, Animals, MESH: Animals, MESH: Mice, Butyrylcholinesterase, 030304 developmental biology, Acetylcholine receptor, Sequence Deletion, Mice, Knockout, 0303 health sciences, [SDV.NEU.NB] Life Sciences [q-bio]/Neurons and Cognition [q-bio.NC]/Neurobiology, General Medicine, Exons, MESH: Acetylcholinesterase, MESH: Sequence Deletion, Acetylcholinesterase, Molecular biology, Respiratory Muscles, medicine.anatomical_structure, chemistry, MESH: Diaphragm, MESH: Receptors, Nicotinic, MESH: Neuromuscular Junction, MESH: Exons, 030217 neurology & neurosurgery
Abstract

At the vertebrate skeletal neuromuscular junction (NMJ), two closely related enzymes can hydrolyze acetylcholine (ACh): acetylcholinesterase (AChE) and butyrylcholinesterase (BChE). Advances in mouse genomics offer new approaches to assess the role of specific cholinesterases involved in neuromuscular transmission (Minic et al., 2003). AChE knockout mice provide a valuable tool for examining the effects of long-term complete and selective abolition of AChE activity (Xie et al., 2000). AChE and BChE genes encode two functional domains--the catalytic domain (exons 2, 3, and 4 of AChE, or exon 2 of BChE) and a C-terminal domain (exon 5 or 6 of AChE, or exon 3 of BChE)--that dictate the targeting of the enzymes (Massoulie, 2002). In mammals, the AChE gene produces three types of coding regions by deleting 5'- splice acceptor sites, which generate proteins; these proteins possess the same catalytic domain associated with distinct C-terminal peptides. AChE subunits of type R (readthrough) produce soluble monomers; they are expressed during development and are thought to be induced in the mouse brain by stress (Kaufer et al., 1998). AChE subunits of type H (hydrophobic) produce GPI-anchored dimers, mainly in blood cells. Subunits of type T (tailed) exist for both AChE and BChE. They represent the predominant AChE variant expressed in cholinergically innervated tissues (muscle and nerve). These subunits generate a variety of quaternary structures, including homomeric oligomers (monomers, dimers, tetramers), as well as hetero-oligomeric assemblies with anchoring proteins ColQ (Krejci et al., 1997) and PRiMA (Perrier et al., 2002). At the NMJ, AChE is clustered by the interaction of the coding sequence of exon 6 with ColQ (Feng et al., 1999). The deletion of exons 5 and 6 in the AChE gene transforms anchored AChE into a soluble enzyme (Camp et al., 2004). The present study was designed to evaluate neuromuscular transmission and nicotinic ACh receptor (nAChR) distribution in muscles from mutant mice with deletions of these two spliced exons (AChE-del-exons-5+6-/-).

Published
2006

19. Synaptic remodeling at the skeletal neuromuscular junction of acetylcholinesterase knockout mice and its physiological relevance

Author

Eric Krejci, Julien Barbier, Arnaud Chatonnet, Jordi Molgó, Emmanuelle Girard, Laboratoire de neurobiologie cellulaire et moléculaire (NBCM), Centre National de la Recherche Scientifique (CNRS), Institut de Neurobiologie Alfred Fessard (INAF), Département de Physiologie Animale, Institut National de la Recherche Agronomique (INRA), Biologie des Jonctions Neuromusculaires Normales et Pathologiques (U686), 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), Département de Physiologie Animale (PA), Différenciation Cellulaire et Croissance (DCC), Institut National de la Recherche Agronomique (INRA)-Université Montpellier 2 - Sciences et Techniques (UM2), and Institut National de la Santé et de la Recherche Médicale (INSERM)

Subjects
Aging, medicine.medical_specialty, Nicotinic acetylcholine receptors, [SDV]Life Sciences [q-bio], Neuromuscular transmission, Motor nerve, Neuromuscular junction, Biology, Toxicology, Mice, 03 medical and health sciences, chemistry.chemical_compound, Nerve terminals, 0302 clinical medicine, Muscle tension, Internal medicine, medicine, BIOLOGIE DU DEVELOPPEMENT, Animals, [INFO]Computer Science [cs], Receptors, Cholinergic, [SDV.NEU] Life Sciences [q-bio]/Neurons and Cognition [q-bio.NC], Muscle, Skeletal, Butyrylcholinesterase, 030304 developmental biology, Acetylcholine receptor, Mice, Knockout, Motor Neurons, 0303 health sciences, General Medicine, Anatomy, Synaptic remodeling, Acetylcholinesterase, Electrophysiology, medicine.anatomical_structure, Nicotinic agonist, Endocrinology, ACETYLCHOLINESTERASE KNOCKOUT MIC, chemistry, Synapses, Acetylcholinesterase knockout mice, NEUROPHYSIOLOGIE, [SDV.NEU]Life Sciences [q-bio]/Neurons and Cognition [q-bio.NC], 030217 neurology & neurosurgery
Abstract

International audience; Acute inhibition of synaptic acetylcholinesterase (AChE) is fatal to normal animals, but AChE-knockout mice (AChE(-/-)) expressing normal levels of butyrylcholinesterase (BChE) could live to adulthood without AChE expression. The present study was under-taken to determine whether compensatory mechanisms occur in the mutant that allow an effective neuromuscular transmission in the chronic absence of AChE. For this we evaluated neuromuscular transmission and the distribution of nicotinic acetylcholine receptors (nAChRs) and motor nerve terminals on isolated nerve-muscle preparations from AChE(-/-) mice. AChE(-/-) hemidiaphragm muscles maintained at 32 degrees C can support muscle twitches, and tetanic contractions during intermittent nerve-stimulation over a wide range of physiological frequencies, even though they develop less force, than age-matched wild-type (AChE(+/+)) muscles. Tetanic fade in AChE(-/-) muscles was temperature-sensitive and more marked at 22 degrees C than at 32 degrees C. Inhibition of BChE by tetraisopropylpyrophosphoramide (Iso-OMPA) intensified tetanic fade in AChE(-/-) muscles, but had no effect on AChE(+/+) muscles, suggesting that BChE plays a protective role in nerve terminals. Skeletal muscles from AChE(-/-) mice adapted to the lack of AChE enzymatic activity by triggering a synaptic remodeling that critically occurred between the second and third week of postnatal development, during synapse elimination. In AChE(-/-) muscles nAChRs distributed in a smaller and fragmented surface area, that mirrored the branching pattern of motor nerve terminals. These findings indicate that the neuromuscular system exhibits a remarkable plasticity and adaptive responses to the chronic absence of AChE activity that has important consequences for the functioning of the neuromuscular junction.

Published
2005
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20. Cholinesterases and the resistance of the mouse diaphragm to the effect of tubocurarine

Author

Tu Nguyen-Huu, Julien Barbier, Jordi Molgó, Alexandre Dobbertin, Philippe Duvaldestin, Eric Krejci, Jasmina Minic, Outters-Lafaye, Michèle, Laboratoire de neurobiologie cellulaire et moléculaire (NBCM), Centre National de la Recherche Scientifique (CNRS), Institut de Neurobiologie Alfred Fessard (INAF), Biologie des Jonctions Neuromusculaires Normales et Pathologiques (U686), Université Paris Descartes - Paris 5 (UPD5)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Centre National de la Recherche Scientifique (CNRS), Service d'anesthésie-réanimation SAMU94-SMUR94 [Mondor], Assistance publique - Hôpitaux de Paris (AP-HP) (AP-HP)-Hôpital Henri Mondor-Université Paris-Est Créteil Val-de-Marne - Paris 12 (UPEC UP12), Assistance publique - Hôpitaux de Paris (AP-HP) (APHP)-Hôpital Henri Mondor-Université Paris-Est Créteil Val-de-Marne - Paris 12 (UPEC UP12), 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 Assistance publique - Hôpitaux de Paris (AP-HP) - Hôpital Henri Mondor - Université Paris-Est Créteil Val-de-Marne - Paris 12 (UPEC UP12)

Subjects
MESH : Neuromuscular Nondepolarizing Agents, Drug Resistance, Muscle Proteins, Tubocurarine, MESH : Dose-Response Relationship, Drug, Receptors, Nicotinic, MESH: Dose-Response Relationship, Drug, chemistry.chemical_compound, Mice, 0302 clinical medicine, 030202 anesthesiology, MESH: Collagen, COLQ, Medicine, MESH: Animals, MESH : Tubocurarine, MESH : Muscle Proteins, biology, musculoskeletal, neural, and ocular physiology, Neuromuscular Blocking Agents, musculoskeletal system, Acetylcholinesterase, Diaphragm (structural system), medicine.anatomical_structure, Biochemistry, MESH: Receptors, Nicotinic, MESH: Drug Resistance, [SDV.NEU]Life Sciences [q-bio]/Neurons and Cognition [q-bio.NC], Collagen, tissues, Acetylcholine, medicine.drug, medicine.medical_specialty, Synaptic cleft, Diaphragm, Neuromuscular Junction, In Vitro Techniques, Neuromuscular junction, MESH : Acetylcholinesterase, 03 medical and health sciences, MESH: Muscle Proteins, Internal medicine, MESH : Mice, Animals, MESH: Tubocurarine, [SDV.NEU] Life Sciences [q-bio]/Neurons and Cognition [q-bio.NC], MESH: Mice, Cholinesterase, MESH : Drug Resistance, MESH : Receptors, Nicotinic, Dose-Response Relationship, Drug, business.industry, MESH: Acetylcholinesterase, Anesthesiology and Pain Medicine, Endocrinology, chemistry, MESH: Diaphragm, MESH : Collagen, biology.protein, MESH : Neuromuscular Junction, MESH : Animals, MESH: Neuromuscular Junction, MESH : Diaphragm, business, MESH: Neuromuscular Nondepolarizing Agents, 030217 neurology & neurosurgery, Neuromuscular Nondepolarizing Agents
Abstract

Background The diaphragm is resistant to competitive neuromuscular blocking agents. Because of the competitive mechanism of action of tubocurarine, the rate of hydrolysis of acetylcholine at the neuromuscular junction may modulate its neuromuscular blocking effect. The authors compared the neuromuscular blocking effect of tubocurarine on isolated diaphragm and extensor digitorum longus (EDL) muscles and quantified the acetylcholinesterase activity in hetero-oligomers. Methods Adult Swiss-Webster and collagen Q-deficient (ColQ) mice were used. The blocking effect of tubocurarine on nerve-evoked muscle twitches was determined in isolated diaphragm and EDL muscles, after inhibition of acetylcholinesterase by fasciculin-1, butyrylcholinesterase by tetraisopropylpyro-phosphoramide, or both acetylcholinesterase and butyrylcholinesterase by neostigmine, and in acetylcholinesterase-deficient ColQ muscles. The different acetylcholinesterase oligomers extracted from diaphragm and EDL muscles were quantified in sucrose gradient. Results The EC50 for tubocurarine to decrease the nerve-evoked twitch response was four times higher in the diaphragm than in the EDL. The activity of the different acetylcholinesterase oligomers was lower in the diaphragm as compared with the EDL. Inhibition of acetylcholinesterase by antagonists resulted in an increased dose of tubocurarine but an unchanged resistance ratio between the diaphragm and the EDL. A similar diaphragmatic resistance was found in ColQ muscles. Conclusion The current study indicates that, despite differences in acetylcholinesterase activity between the diaphragm and EDL, the diaphragmatic resistance to tubocurarine cannot be explained by the different rate of acetylcholine hydrolysis in the synaptic cleft.

Published
2005

21. Neuromuscular transmission and synaptic remodeling at the neuromuscular junction of knockout mice with partial deletions in the acetylcholinesterase gene

Author

Girard, Emmanuelle, Camp, Shelley, Taylor, Palmer, Krejci, E., Molgó, Jordi, Laboratoire de neurobiologie cellulaire et moléculaire (NBCM), Centre National de la Recherche Scientifique (CNRS), Department of pharmacology, University of California [San Diego] (UC San Diego), University of California-University of California, Biologie des Jonctions Neuromusculaires Normales et Pathologiques (U686), 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 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
Butyrylcholinesterase, [SDV.NEU.NB]Life Sciences [q-bio]/Neurons and Cognition [q-bio.NC]/Neurobiology, [SDV.NEU.NB] Life Sciences [q-bio]/Neurons and Cognition [q-bio.NC]/Neurobiology, Acetylcholinesterase, ComputingMethodologies_GENERAL, Genomics, Neuromuscular transmission
Abstract

Poster

Published
2005

22. Acetylcholinesterase and molecular interactions at the neuromuscular junction

Author

Claire Legay, Jean Cartaud, Annie Cartaud, M. Guerra, Biologie des Jonctions Neuromusculaires Normales et Pathologiques (U686), 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 Jacques Monod (IJM (UMR_7592)), Université Paris Diderot - Paris 7 (UPD7)-Centre National de la Recherche Scientifique (CNRS), Kropfinger, Antonia, 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 Jacques Monod (IJM), and Université Paris Diderot - Paris 7 (UPD7) - Centre National de la Recherche Scientifique (CNRS)

Subjects
medicine.medical_specialty, Neuromuscular Junction, Perlecan, [SDV.BC.IC] Life Sciences [q-bio]/Cellular Biology/Cell Behavior [q-bio.CB], Neurotransmission, Toxicology, Neuromuscular junction, 03 medical and health sciences, chemistry.chemical_compound, Mice, 0302 clinical medicine, Internal medicine, COLQ, [SDV.BC.IC]Life Sciences [q-bio]/Cellular Biology/Cell Behavior [q-bio.CB], medicine, Myocyte, Animals, Humans, 030304 developmental biology, Acetylcholine receptor, 0303 health sciences, biology, Cell Differentiation, General Medicine, Acetylcholinesterase, Cell biology, medicine.anatomical_structure, Endocrinology, chemistry, biology.protein, Basal lamina, Collagen, 030217 neurology & neurosurgery, Protein Binding
Abstract

The efficiency and the tight control of neurotransmission require the accumulation of synaptic proteins in discrete domains. In neuromuscular junctions, the main form of acetylcholinesterase (AChE) is a hetero-oligomer in which the catalytic subunits are associated to a specific collagen, ColQ. This structural protein is responsible for the insertion and the accumulation of AChE in the synaptic basal lamina. We have analyzed the time-course of acetylcholinesterase and acetylcholine receptors (AChR) mRNAs during mouse muscle cell differentiation in culture. In parallel, we have visualized the formation of AChE and AChR aggregates. We show that AChR clusters form first which correlates with high gamma-subunit mRNA levels. Then, AChE clusters appear with the onset of contraction and correlate with a dramatic increase in AChE, ColQ1 and ColQ1A mRNA levels in muscle cells. At that stage, AChR gamma-subunit levels drop while the expression level of epsilon-subunits increase. AChE aggregates are organized by a ternary complex, which involves direct interactions between ColQ, perlecan and MuSK.

Published
2005

23. Synaptic efficacy and remodelling at the neuromuscular junction of knockout mice deficient in acetylcholinesterase

Author

Emmanuelle Girard, Arnaud Chatonnet, Jordi Molgó, Eric Krejci, Shelley Camp, Palmer Taylor, Julien Barbier, Laboratoire de neurobiologie cellulaire et moléculaire (NBCM), Centre National de la Recherche Scientifique (CNRS), Institut de Neurobiologie Alfred Fessard (INAF), Department of pharmacology, University of California [San Diego] (UC San Diego), University of California-University of California, Département de Physiologie Animale, Institut National de la Recherche Agronomique (INRA), Biologie des Jonctions Neuromusculaires Normales et Pathologiques (U686), Université Paris Descartes - Paris 5 (UPD5)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Centre National de la Recherche Scientifique (CNRS), Département de Physiologie Animale (PA), 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
0303 health sciences, General Neuroscience, Synaptic efficacy, Acetylcholinesterase, Neuromuscular junction, Cell biology, 03 medical and health sciences, chemistry.chemical_compound, 0302 clinical medicine, medicine.anatomical_structure, chemistry, Physiology (medical), Knockout mouse, medicine, [SDV.NEU]Life Sciences [q-bio]/Neurons and Cognition [q-bio.NC], [SDV.NEU] Life Sciences [q-bio]/Neurons and Cognition [q-bio.NC], 030217 neurology & neurosurgery, 030304 developmental biology
Published
2004

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