Asynchronous assembly of the acetylcholine receptor and of the 43-kD nu1 protein in the postsynaptic membrane of developing Torpedo marmorata electrocyte.
AuthorsKordeli E, Cartaud J, Nghiêm HO, Devillers-Thiéry A, Changeux JP
JournalJ Cell Biol
PubMed ID2642909
'The assembly of the nicotinic acetylcholine receptor (AchR) and the 43-kD protein (v1), the two major components of the post synaptic membrane of the electromotor synapse, was followed in Torpedo marmorata electrocyte during embryonic development by immunocytochemical methods. At the first developmental stage investigated (45-mm embryos), accumulation of AchR at ... More
Changes in architecture of the Golgi complex and other subcellular organelles during myogenesis.
AuthorsRalston E
JournalJ Cell Biol
PubMed ID7678420
'Myogenesis involves changes in both gene expression and cellular architecture. Little is known of the organization, in muscle in vivo, of the subcellular organelles involved in protein synthesis despite the potential importance of targeted protein synthesis for formation and maintenance of functional domains such as the neuromuscular junction. A panel ... More
Time-dependent binding of paramagnetic and fluorescent hydrophobic ions to the acetylcholine receptor from Torpedo.
AuthorsHartsel SC, Moore CR, Raines DE, Cafiso DS
JournalBiochemistry
PubMed ID2820466
'In receptor-rich vesicles isolated from Torpedo, paramagnetic or fluorescent phosphonium ions bind to both the acetylcholine receptor (AcChR) and the receptor membrane. When added to receptor vesicles, two to three phosphoniums undergo a slow time-dependent binding to the AcChR. The presence of agonist increases the rate but not the extent ... More
Myoseverin, a microtubule-binding molecule with novel cellular effects.
'A new microtubule-binding molecule, myoseverin, was identified from a library of 2,6,9-trisubstituted purines in a morphological differentiation screen. Myoseverin induces the reversible fission of multinucleated myotubes into mononucleated fragments. Myotube fission promotes DNA synthesis and cell proliferation after removal of the compound and transfer of the cells to fresh growth ... More
The small leucine-rich repeat proteoglycan biglycan binds to alpha-dystroglycan and is upregulated in dystrophic muscle.
AuthorsBowe MA, Mendis DB, Fallon JR
JournalJ Cell Biol
PubMed ID10684260
'The dystrophin-associated protein complex (DAPC) is necessary for maintaining the integrity of the muscle cell plasma membrane and may also play a role in coordinating signaling events at the cell surface. The alpha-/beta-dystroglycan subcomplex of the DAPC forms a critical link between the cytoskeleton and the extracellular matrix. A ligand ... More
Membrane targeting and stabilization of sarcospan is mediated by the sarcoglycan subcomplex.
'The dystrophin-glycoprotein complex (DGC) is a multisubunit complex that spans the muscle plasma membrane and forms a link between the F-actin cytoskeleton and the extracellular matrix. The proteins of the DGC are structurally organized into distinct subcomplexes, and genetic mutations in many individual components are manifested as muscular dystrophy. We ... More
Identification of nicotinic acetylcholine receptors on lymphocytes in the periphery as well as thymus in mice.
AuthorsToyabe S, Iiai T, Fukuda M, Kawamura T, Suzuki S, Uchiyama M, Abo T
JournalImmunology
PubMed ID9415027
'The existence of nicotinic acetylcholine receptors (nAChR) on lymphocytes remains controversial. We attempted to show the existence of nAChR on murine lymphocytes. The intraperitoneal injection of nicotine induced the lymphocytosis in the spleen on day 3. Although freshly isolated lymphocytes bound small quantities of fluorescein isothiocyanate (FITC)-conjugated alpha-bungarotoxin (alpha BuTx), ... More
Nicotinic acetylcholine receptor alpha7 subunit is an essential regulator of inflammation.
AuthorsWang H, Yu M, Ochani M, Amella CA, Tanovic M, Susarla S, Li JH, Wang H, Yang H, Ulloa L, Al-Abed Y, Czura CJ, Tracey KJ
JournalNature
PubMed ID12508119
'Excessive inflammation and tumour-necrosis factor (TNF) synthesis cause morbidity and mortality in diverse human diseases including endotoxaemia, sepsis, rheumatoid arthritis and inflammatory bowel disease. Highly conserved, endogenous mechanisms normally regulate the magnitude of innate immune responses and prevent excessive inflammation. The nervous system, through the vagus nerve, can inhibit significantly ... More
Activity-dependent fluorescent staining and destaining of living vertebrate motor nerve terminals.
AuthorsBetz WJ, Mao F, Bewick GS
JournalJ Neurosci
PubMed ID1371312
'Living motor nerve terminals from several species can be stained in an activity-dependent fashion by certain styryl dyes, such as RH414, RH795, and a new dye, FM1-43, which can be imaged independently of the others. The dyes evidently become trapped within recycled synaptic vesicles. In frog cutaneus pectoris muscle, bright ... More
Fluorescently labelled Na+ channels are localized and immobilized to synapses of innervated muscle fibres.
AuthorsAngelides KJ
JournalNature
PubMed ID2422557
'Segregation of voltage-dependent sodium channels to the hillock of motoneurones and nodes of Ranvier in myelinated axons is crucial for conduction of the nerve impulse. Much less is known, however, about the distribution of voltage-dependent Na+ channels on muscle fibres. Recently, Beam et al. have shown that Na+ channels are ... More
Neural agrin controls acetylcholine receptor stability in skeletal muscle fibers.
'At mammalian neuromuscular junctions (NMJs), innervation induces and maintains the metabolic stability of acetylcholine receptors (AChRs). To explore whether neural agrin may cause similar receptor stabilization, we injected neural agrin cDNA of increasing transfection efficiencies into denervated adult rat soleus (SOL) muscles. As the efficiency increased, the amount of recombinant ... More
Nerve sprouting in innervated adult skeletal muscle induced by exposure to elevated levels of insulin-like growth factors.
AuthorsCaroni P, Grandes P
JournalJ Cell Biol
PubMed ID2157718
'Partial denervation or paralysis of adult skeletal muscle is followed by nerve sprouting, probably due to release of diffusible sprout-inducing activity by inactive muscle. Insulin-like growth factors (IGF1 and IFG2) are candidates for muscle-derived sprouting activity, because (a) they induce neurite growth from peripheral neurons in vitro; and (b) their ... More
Control of the autoimmune response by type 2 nitric oxide synthase.
AuthorsShi FD, Flodström M, Kim SH, Pakala S, Cleary M, Ljunggren HG, Sarvetnick N
JournalJ Immunol
PubMed ID11509651
'Immune defense against pathogens often requires NO, synthesized by type 2 NO synthase (NOS2). To discern whether this axis could participate in an autoimmune response, we immunized NOS2-deficient mice with the autoantigen acetylcholine receptor, inducing muscle weakness characteristic of myasthenia gravis, a T cell-dependent Ab-mediated autoimmune disease. We found that ... More
Histochemical staining of the acetylcholine receptor, acetylcholinesterase, and the axon terminal.
AuthorsBjornskov EK, Norris FH, Mower-Kuby J
JournalMuscle Nerve
PubMed ID7070395
Denervated skeletal muscle fibers develop discrete patches of high acetylcholine receptor density.
AuthorsKo PK, Anderson MJ, Cohen MW
JournalScience
PubMed ID850796
Denervated skeletal muscle fibers of mice develop discrete patches of high acetylcholine receptor density. The patches vary in size from less than 1 micrometer up to 30 micrometers, depending on the muscle and the period of denervation. Within the patches the acetylcholine receptor density is some 20 times greater than ... More
The calcium channel and the organization of the presynaptic transmitter release face.
AuthorsStanley EF
JournalTrends Neurosci
PubMed ID9292969
Calcium influx through ion channels located on the release face of the presynaptic nerve terminal gates the release of neurotransmitters by the fusion of the secretory vesicle and the discharge of its contents. Recently, several lines of research have indicated that the relationship between the Ca2+ channel and the release ... More
Binding pattern of alpha-bungarotoxin on horizontal cells of a marine teleost retina.
AuthorsDeplano S
JournalJ Comp Neurol
PubMed ID3209745
A conjugate of alpha-bungarotoxin and a fluorescent marker (fluorescein isothiocyanate) has been used to localize "nicotinic" acetylcholine receptors on neurons in the outer plexiform layer of marine teleost retina. Toxin binding was confined to bipolar cell dendrites and to intermediate horizontal cells. The arrangement of labeled horizontal cells appears irregular ... More
Rapid adhesion of nerve cells to muscle fibers from adult rats is mediated by a sialic acid-binding receptor.
AuthorsBischoff R
JournalJ Cell Biol
PubMed ID3711146
Single viable muscle fibers isolated from adult rats by collagenase digestion rapidly bind dissociated spinal neurons or PC-12 cells but not a variety of other cells tested. The adhesion process is calcium-independent, temperature-sensitive, and is not blocked by pretreating cells with inhibitors of energy metabolism or actin polymerization. Adhesion is ... More
Fluorophore-assisted light inactivation produces both targeted and collateral effects on N-type calcium channel modulation in rat sympathetic neurons.
AuthorsGuo J, Chen H, Puhl HL, Ikeda SR,
JournalJ Physiol
PubMed ID16873413
Fluorophore-assisted light inactivation (FALI) is a method to inactivate specific proteins on a time scale of seconds to minutes using either diffuse or coherent light. Here we examine a novel FALI modality that utilizes a fluorescein-conjugated polypeptide, alpha-bungarotoxin (BTX) and a 13 amino acid BTX-binding site engineered into the N-terminus ... More
Association of Rab3A with synaptic vesicles at late stages of the secretory pathway.
AuthorsMatteoli M, Takei K, Cameron R, Hurlbut P, Johnston PA, Südhof TC, Jahn R, De Camilli P
JournalJ Cell Biol
PubMed ID1655810
Rab3A is a small GTP-binding protein highly concentrated on synaptic vesicles. Like other small GTP-binding proteins it is thought to cycle between a soluble and a membrane-associated state. To determine at which stage of the life cycle of synaptic vesicles rab3A is associated with their membranes, the localization of the ... More
Selective labeling of alpha-bungarotoxin with fluorescein isothiocyanate and its use for the study of toxin-acetylcholine receptor interactions.
AuthorsGarcia-Borron JC, Chinchetru MA, Martinez-Carrion M
JournalJ Protein Chem
PubMed ID2127357
The main product of the reaction of fluorescein isothiocyanate (FITC) and bungarotoxin (Bgt) under near stoichiometric conditions is a monofluorescein derivative preferentially labeled at Lys 26, a highly conserved residue known to be involved in the binding (McDaniel, C.S., Manshouri, T., and Atassi, M.Z. (1987) J. Prot. Chem. 6, 455-461; ... More
Distribution of Ca2+ channels on frog motor nerve terminals revealed by fluorescent omega-conotoxin.
AuthorsCohen MW, Jones OT, Angelides KJ
JournalJ Neurosci
PubMed ID1707093
Tetramethylrhodamine-conjugated omega-conotoxin was used as a fluorescent stain (Jones et al., 1989) to determine the spatial distribution of voltage-gated Ca2+ channels along frog motor nerve terminals. Like native omega-conotoxin, the fluorescent toxin blocked neuromuscular transmission irreversibly. The fluorescent staining was confined to the neuromuscular junction and consisted of a series ... More
Postsynaptic abnormalities at the neuromuscular junctions of utrophin-deficient mice.
AuthorsDeconinck AE, Potter AC, Tinsley JM, Wood SJ, Vater R, Young C, Metzinger L, Vincent A, Slater CR, Davies KE
JournalJ Cell Biol
PubMed ID9049253
Utrophin is a dystrophin-related cytoskeletal protein expressed in many tissues. It is thought to link F-actin in the internal cytoskeleton to a transmembrane protein complex similar to the dystrophin protein complex (DPC). At the adult neuromuscular junction (NMJ), utrophin is precisely colocalized with acetylcholine receptors (AChRs) and recent studies have ... More
Fluorescent staining of acetylcholine receptors in vertebrate skeletal muscle.
AuthorsAnderson MJ, Cohen MW
JournalJ Physiol
PubMed ID4133039
1. alpha-Bungarotoxin was labelled with fluorescent dyes and used as a stain for visualizing the distribution of acetylcholine receptors in vertebrate skeletal muscle fibres.2. Dye-toxin conjugates had the same pharmacological properties as native toxin, but their potencies were lower.3. Fluorescent staining was examined in teased muscle fibres. The stain was ... More
Presence of a protein immunologically related to lamin B in the postsynaptic membrane of Torpedo marmorata electrocyte.
AuthorsCartaud A, Courvalin JC, Ludosky MA, Cartaud J
JournalJ Cell Biol
PubMed ID2677028
The Torpedo electrocyte is a flattened syncytium derived from skeletal muscle, characterized by two functionally distinct plasma membrane domains. The electrocyte is filled up with a transversal network of intermediate filaments (IF) of desmin which contact in an end-on fashion both sides of the cell. In this work, we show ... More
Calcitonin gene-related peptide: possible role in formation and maintenance of neuromuscular junctions.
AuthorsSala C, Andreose JS, Fumagalli G, Lømo T
JournalJ Neurosci
PubMed ID7823160
The expression and content of calcitonin gene-related peptide (CGRP) and secretogranin II (SgII) in adult rat motor neurons were examined by in situ hybridization, Northern blot analysis, and immunocytochemistry. Normal motor nerve terminals did not contain detectable CGRP or SgII. Ten to 15 days after a peripheral nerve crush about ... More
MAGI-1c: a synaptic MAGUK interacting with muSK at the vertebrate neuromuscular junction.
The muscle-specific receptor tyrosine kinase (MuSK) forms part of a receptor complex, activated by nerve-derived agrin, that orchestrates the differentiation of the neuromuscular junction (NMJ). The molecular events linking MuSK activation with postsynaptic differentiation are not fully understood. In an attempt to identify partners and/or effectors of MuSK, cross-linking and ... More
beta-Spectrin is colocalized with both voltage-gated sodium channels and ankyrinG at the adult rat neuromuscular junction.
AuthorsWood SJ, Slater CR
JournalJ Cell Biol
PubMed ID9456326
Voltage-gated sodium channels (VGSCs) are concentrated in the depths of the postsynaptic folds at mammalian neuromuscular junctions (NMJs) where they facilitate action potential generation during neuromuscular transmission. At the nodes of Ranvier and the axon hillocks of central neurons, VGSCs are associated with the cytoskeletal proteins, beta-spectrin and ankyrin, which ... More
Polarized fluorescence photobleaching recovery for measuring rotational diffusion in solutions and membranes.
AuthorsVelez M, Axelrod D
JournalBiophys J
PubMed ID3382712
A variation of fluorescence photobleaching recovery (FPR) suitable for measuring the rate of rotational molecular diffusion in solution and cell membranes is presented in theory and experimental practice for epi-illumination microscopy. In this technique, a brief flash of polarized laser light creates an anisotropic distribution of unbleached fluorophores which relaxes ... More
A protein homologous to the Torpedo postsynaptic 58K protein is present at the myotendinous junction.
AuthorsChen Q, Sealock R, Peng HB
JournalJ Cell Biol
PubMed ID2112550
The 58K protein is a peripheral membrane protein enriched in the acetylcholine receptor (AChR)-rich postsynaptic membrane of Torpedo electric organ. Because of its coexistence with AChRs in the postsynaptic membrane in both electrocytes and skeletal muscle, it is thought to be involved in the formation and maintenance of AChR clusters. ... More
Aggregates of acetylcholine receptors are associated with plaques of a basal lamina heparan sulfate proteoglycan on the surface of skeletal muscle fibers.
AuthorsAnderson MJ, Fambrough DM
JournalJ Cell Biol
PubMed ID6226669
Hybridoma techniques have been used to generate monoclonal antibodies to an antigen concentrated in the basal lamina at the Xenopus laevis neuromuscular junction. The antibodies selectively precipitate a high molecular weight heparan sulfate proteoglycan from conditioned medium of muscle cultures grown in the presence of [35S]methionine or [35S]sulfate. Electron microscope ... More
The lipid bilayer of acetylcholine receptor clusters of cultured rat myotubes is organized into morphologically distinct domains.
AuthorsScher MG, Bloch RJ
JournalExp Cell Res
PubMed ID2055278
We have studied the composition and organization of the lipid bilayer at the large, substrate-associated clusters of acetylcholine receptors (AChR) that form in cultured rat myotubes. These clusters have a characteristic morphology consisting of alternating linear domains of AChR-rich and AChR-poor membrane, the latter involved in attaching the myotube to ... More
The role of heparin-binding growth-associated molecule (HB-GAM) in the postsynaptic induction in cultured muscle cells.
AuthorsPeng HB, Ali AA, Dai Z, Daggett DF, Raulo E, Rauvala H
JournalJ Neurosci
PubMed ID7722643
The heparan sulfate proteoglycan (HSPGs) is a components of the extracellular matrix of skeletal muscle that is concentrated at the neuromuscular junction (NMJ). Recent studies have suggested that HSPG, together with its bound peptide growth factors, plays important roles in autocrine or paracrine types of regulation of cell growth and ... More
Effects of innervation on the distribution of acetylcholine receptors in regenerating skeletal muscles of adult chickens.
AuthorsIshikawa Y, Arakaki A, Shimizu N, Ibaraki K, Tanaka S
JournalDev Biol
PubMed ID3334713
In order to determine the roles of nerves in the formation of clusters of acetylcholine receptors (AChRs) during synaptogenesis, we examined the distribution of AChRs in denervated, nerve-transplanted (neurotized) muscles and in regenerated skeletal muscles of adult chickens by fluorescence microscopy using curaremimetic toxins. In the denervated muscles, many extrajunctional ... More
N-CAM at the vertebrate neuromuscular junction.
AuthorsRieger F, Grumet M, Edelman GM
JournalJ Cell Biol
PubMed ID3891761
We have detected the neural cell adhesion molecule, N-CAM, at nerve-muscle contacts in the developing and adult mouse diaphragm. Whereas we found N-CAM staining with fluorescent antibodies consistently to overlap with the pattern of alpha-bungarotoxin staining at nerve-muscle contacts both during development and in the adult, we observed N-CAM staining ... More
The formation of terminal fields in the absence of competitive interactions among primary motoneurons in the zebrafish.
AuthorsLiu DW, Westerfield M
JournalJ Neurosci
PubMed ID2269893
To make specific synaptic connections, projection neurons extend neurites to regions containing appropriate targets, then form synapses with the correct type and number of target cells. To investigate the mechanisms controlling this process, we have studied the formation of motoneuronal terminal fields in live zebrafish embryos. The primary motoneurons of ... More