Precise localization of alpha7 nicotinic acetylcholine receptors on glutamatergic axon terminals in the rat ventral tegmental area.
AuthorsJones IW, Wonnacott S
JournalJ Neurosci
PubMed ID15601930
'Alpha7 neuronal nicotinic acetylcholine receptors (nAChRs) constitute one of the predominant nAChR subtypes in the mammalian brain. Within the ventral tegmental area (VTA), nicotine application, paired with postsynaptic stimulation, contributes to a form of long-term potentiation, an effect attributed to presynaptic alpha7 nAChRs on glutamatergic afferents (Mansvelder and McGehee, 2000). ... More
Imaging of receptor trafficking by using alpha-bungarotoxin-binding-site-tagged receptors.
AuthorsSekine-Aizawa Y, Huganir RL,
JournalProc Natl Acad Sci U S A
PubMed ID15563595
'alpha-Amino-3-hydroxy-5-methyl-4-isoxazole-propionate (AMPA) receptors mediate excitatory synaptic transmission and are dynamically regulated during synaptic plasticity in the CNS. The membrane trafficking of AMPA receptors to synapses is critical for the regulation of the efficacy of excitatory synaptic transmission. Direct imaging of AMPA receptors in various cell compartments is important to dissecting ... More
alpha-Bungarotoxin binding sites in rat hippocampal and cortical cultures: initial characterisation, colocalisation with alpha 7 subunits and up-regulation by chronic nicotine treatment.
AuthorsBarrantes GE, Rogers AT, Lindstrom J, Wonnacott S
JournalBrain Res
PubMed ID7749744
'High density neuronal cultures from rat E18 hippocampus and cortex have been characterised with respect to cholinergic binding sites. No specific binding of [3H]nicotine or [3H]cytisine to live cells in situ was detected although the limit for detection was estimated to be 30 fmol/mg protein. Muscarinic binding sites labelled with ... More
The short-neurotoxin-binding regions on the alpha-chain of human and Torpedo californica acetylcholine receptors.
AuthorsRuan KH, Stiles BG, Atassi MZ
JournalBiochem J
PubMed ID2012611
'The continuous regions for short-neurotoxin binding on the alpha-chains of Torpedo californica (electric ray) and human acetylcholine receptors (AChR) were localized by reaction of 125I-labelled cobrotoxin (Cot) and erabutoxin b (Eb) with synthetic overlapping peptides spanning the entire extracellular part of the respective alpha-chains. On Torpedo AChR, five Cot-binding regions ... More
Actions of vesamicol on an alpha-bungarotoxin-sensitive neuronal nicotinic acetylcholine receptor.
AuthorsBuckingham SD, Lummis SC, Balk ML, Schroeder M, Sattelle DB
JournalJ Exp Biol
PubMed ID8228781
'Electrophysiology and binding studies were used to determine the actions of vesamicol [2-(4-phenylpiperidino)cyclohexanol (AH5183)] on an alpha-bungarotoxin-sensitive, neuronal nicotinic acetylcholine receptor in the nervous system of the cockroach, Periplaneta americana. Electrophysiological studies on an identified motor neurone revealed a reversible blocking action of (+/-)-vesamicol on the response to ionophoretically applied ... 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
Nicotine receptors in the mammalian brain.
AuthorsLuetje CW, Patrick J, Séguéla P
JournalFASEB J
PubMed ID2197155
'Nicotine is a drug of abuse that presumably exerts its psychoactive effect through its interactions with nicotine binding sites in the central nervous system. Among its potential sites of action are the neuronal nicotinic acetylcholine receptors and the neuronal alpha-bungarotoxin binding sites. In this review we focus on the neuronal ... More
Measurement of junctional acetylcholine receptors in myasthenia gravis: clinical correlates.
AuthorsPestronk A, Drachman DB, Self SG
JournalMuscle Nerve
PubMed ID4058469
'We measured the number of acetylcholine receptors (AChRs) at neuromuscular junctions (NMJs) in motor point biopsies from 76 patients with myasthenia gravis (MG) and 49 control subjects, using an alpha-bungarotoxin binding technique. The mean number of AChRs/NMJ was significantly lower (P less than 0.001) in muscles of MG patients (deltoid ... More
Acetylcholine and local anesthetic binding to Torpedo nicotinic postsynaptic membranes after removal of nonreceptor peptides.
AuthorsNeubig RR, Krodel EK, Boyd ND, Cohen JB
JournalProc Natl Acad Sci U S A
PubMed ID34154
'After alkaline extraction, purified subsynaptic fragments isolated from Torpedo electric tissue exhibit on sodium dodecyl sulfate/polyacrylamide gel electrophoresis predominant peptides of apparent Mr 41,000, 50,000, and 65,000 (i.e., the peptides characteristic of the nicotinic receptor purified and isolated in detergent solutions). The peptide of Mr 43,000 that is also found ... More
Interaction of a semirigid agonist with Torpedo acetylcholine receptor.
AuthorsKawai H, Cao L, Dunn SM, Dryden WF, Raftery MA
JournalBiochemistry
PubMed ID10747774
'The binding of the semirigid agonist [(3)H]arecolone methiodide to the Torpedo nicotinic acetylcholine receptor has been correlated with its functional properties measured both in flux studies with Torpedo membrane vesicles and by single-channel analysis after reconstitution in giant liposomes. Under both equilibrium and preequilibrium conditions, the binding of arecolone methiodide ... More
Cholinergic function and alpha-bungarotoxin binding in PC12 cells.
AuthorsKemp G, Edge M
JournalMol Pharmacol
PubMed ID2890091
'The cell line PC12, derived from an adrenal chromaffin cell tumor, expresses both ganglionic (C6) acetylcholine receptors (nAChR) and an alpha-bungarotoxin (BGT) binding protein of unknown function. We measured nicotinic Na+ fluxes of 180-260 nmol/mg protein X min and 0.35-0.8 pmol [125I]BGT binding sites/mg protein; 45-65% of the [125I]BGT binding ... More
The effects of alpha- and beta-neurotoxins from the venoms of various snakes on transmission in autonomic ganglia.
AuthorsChiappinelli VA, Cohen JB, Zigmond RE
JournalBrain Res
PubMed ID6261879
'We have previously shown that certain commercially available lots of alpha-bungarotoxin block transmission in ciliary and choroid neurons of both pigeon and chicken ciliary ganglia at a concentration of 10 microgram/ml (1.2 microM). The blockade is antagonized by pre-incubation with 100 microM tubocurarine. Further evidence that this blockade is produced ... More
Membrane tethering enables an extracellular domain of the acetylcholine receptor alpha subunit to form a heterodimeric ligand-binding site.
AuthorsWang ZZ, Hardy SF, Hall ZW
JournalJ Cell Biol
PubMed ID8909552
'The first step of assembly of the nicotinic acetylcholine receptor (AChR) of adult skeletal muscle is the specific association of the alpha subunit with either delta or epsilon subunits to form a heterodimer with a ligand-binding site. Previous experiments have suggested that het erodimer formation in the ER arises from ... More
Labeling of functionally sensitive sulfhydryl-containing domains of acetylcholine receptor from Torpedo californica membranes.
AuthorsClarke JH, Martinez-Carrion M
JournalJ Biol Chem
PubMed ID3733702
'N-(1-Pyrene)maleimide, a fluorescent, lipophilic, alkylating agent, was used as a probe for the nicotinic acetylcholine receptor (AChR). Preincubation with N-(1-pyrene)maleimide under nonreducing conditions inhibits agonist-induced cation permeability of AChR-enriched membranes. This inhibition is dependent on the concentration of N-(1-pyrene)maleimide used. This correlation was also exhibited by resonance energy transfer of ... More
Identification and functional characterization of a new agonist site on nicotinic acetylcholine receptors of cultured hippocampal neurons.
AuthorsPereira EF, Reinhardt-Maelicke S, Schrattenholz A, Maelicke A, Albuquerque EX
JournalJ Pharmacol Exp Ther
PubMed ID8510023
'Electrophysiological and biochemical techniques were used to demonstrate that alpha-bungarotoxin-, methyllycaconitine-sensitive neuronal nicotinic acetylcholine receptors (nAChRs) can be activated via a novel agonist site(s). The residue proposed to be essential to this site is the amino acid Lys-125 of the receptor alpha subunit. In outside-out patches excised from cultured hippocampal ... More
The nicotinic blocking agents d-tubocurare and alpha-bungarotoxin save motoneurons from naturally occurring death in the absence of neuromuscular blockade.
AuthorsHory-Lee F, Frank E
JournalJ Neurosci
PubMed ID7472408
'Motoneurons undergo a phase of target-dependent cell death during development. In chick embryos, motoneuronal death is blocked by the application of the nicotinic antagonists d-tubocurare (dTC) or alpha-bungarotoxin (alpha BTX). Paralytic doses of these drugs also increase intramuscular nerve branch formation. To investigate the possibility that a neuronal rather than ... More
An emerging pharmacology of peptide toxins targeted against potassium channels.
AuthorsMoczydlowski E, Lucchesi K, Ravindran A
JournalJ Membr Biol
PubMed ID2464066
'Voltage-dependent ion channels are a difficult class of proteins to approach biochemically. Many such channels are present at low density in relevant tissues and exist as multiple subtypes that can be distinguished electrophysiologically. In particular, K channels appear to be a diverse family of proteins characterized by many different conductance ... More
Thermotropic and dynamic characterization of interactions of acylated alpha-bungarotoxin with phospholipid bilayer membranes.
AuthorsBabbitt B, Huang L, Freire E
JournalBiochemistry
PubMed ID6091732
'The interactions of palmitoyl-alpha-bungarotoxin (PBGT) with dipalmitoylphosphatidylcholine (DPPC) bilayers have been studied by using high-sensitivity differential scanning calorimetry together with steady-state and time-resolved phosphorescence and fluorescence spectroscopy. The incorporation of PBGT into large single lamellar vesicles causes a decrease in the phospholipid phase transition temperature (Tm), a broadening of the ... More
Profile of the alpha-bungarotoxin-binding regions on the extracellular part of the alpha-chain of Torpedo californica acetylcholine receptor.
AuthorsMulac-Jericevic B, Atassi MZ
JournalBiochem J
PubMed ID3435488
'The continuous alpha-neurotoxin-binding regions on the extracellular part (residues 1-210) of the alpha-chain of Torpedo californica acetylcholine receptor were localized by reaction of 125I-labelled alpha-bungarotoxin with synthetic overlapping peptides spanning this entire part of the chain. The specificity of the binding was confirmed by inhibition with unlabelled toxin and, for ... More
Thymopoietin, a thymic polypeptide, specifically interacts at neuronal nicotinic alpha-bungarotoxin receptors.
AuthorsQuik M, Afar R, Audhya T, Goldstein G
JournalJ Neurochem
PubMed ID2769271
'alpha-Bungarotoxin (alpha-BGT), a snake venom polypeptide, interacts potently and specifically with a nicotinic receptor population in neuronal tissue. However, the identity of this site is unclear, because, unlike at the neuromuscular junction and in electroplax, in nervous tissue the toxin does not block nicotinic cholinergic responses. Therefore, we sought endogenous ... More
Ligand binding sites and subunit interactions of Torpedo californica acetylcholine receptor.
AuthorsWitzemann V, Raftery M
JournalBiochemistry
PubMed ID687601
'A [3H]bisazido derivative of ethidium bromide was synthesized to identify sites of interaction of ethidium with the acetylcholine receptor from Torpedo californica and to aid in localization of ligand binding sites. For purified solubilized acetylcholine receptor it was shown (a) that the photolabel was competitive with ethidium bromide, (b) dodecyl ... More
Nicotinic receptors that bind alpha-bungarotoxin on neurons raise intracellular free Ca2+.
AuthorsVijayaraghavan S, Pugh PC, Zhang ZW, Rathouz MM, Berg DK
JournalNeuron
PubMed ID1310863
'Many populations of vertebrate neurons have a membrane component that binds alpha-bungarotoxin and cholinergic ligands. Despite the abundance of this component and its similarities to nicotinic receptors, its function has remained controversial. Using a fluorescence assay, we show here that activation of the component elevates the intracellular concentration of free ... More
Quantitation of cation transport by reconstituted membrane vesicles containing purified acetylcholine receptor.
AuthorsWu WC, Moore HP, Raftery MA
JournalProc Natl Acad Sci U S A
PubMed ID6940146
'A stopped-flow spectroscopic technique was used to study the kinetics of ion transport by reconstituted membrane preparations containing purified acetylcholine receptor. Influx of thallium (I) into membrane vesicles was monitored as a decrease, due to quenching by the thallous ion, in the fluorescence of an entrapped fluorophore. In a reproducible ... More
Eserine and other tertiary amine interactions with Torpedo acetylcholine receptor postsynaptic membrane vesicles.
AuthorsKawai H, Carlson BJ, Okita DK, Raftery MA
JournalBiochemistry
PubMed ID9890891
'Interaction of the tertiary amines, arecolone, eserine (physostigmine), (+)-epibatidine, and (+/-)-epibatidine, with Torpedo nicotinic acetylcholine receptor-enriched membrane vesicles was investigated to characterize their action on the receptor, using stopped-flow thallium (I)-flux spectrofluorimetry. Arecolone, (+)-epibatidine, and (+/-)-epibatidine were agonists with activation constants of 390, 19, and 39 microM, respectively. Eserine was ... More
Neurons can maintain multiple classes of nicotinic acetylcholine receptors distinguished by different subunit compositions.
AuthorsConroy WG, Berg DK
JournalJ Biol Chem
PubMed ID7876208
'Although 10 genes have been cloned encoding putative subunits of neuronal nicotinic acetylcholine receptors, little is known about the variety or subunit composition of such receptors expressed by individual neurons. Chick ciliary ganglion neurons express five of the known genes and assemble a class of synaptic-type receptors collectively containing gene ... More
Regenerating muscle fibers induce directional sprouting from nearby nerve terminals: studies in living mice.
Authorsvan Mier P, Lichtman JW
JournalJ Neurosci
PubMed ID8083762
'The principal aim of this work was to better understand how regenerating muscle fibers become innervated in adult animals. To induce muscle regeneration, individual identified muscle fibers in a mouse were damaged with a laser focused through a microscope. The muscle fiber that degenerated and the muscle fiber that was ... More
Chemical modification of arginine residues in alpha-bungarotoxin.
AuthorsLin SR, Chang CC
JournalBiochim Biophys Acta
PubMed ID1390930
'The reaction of alpha-bungarotoxin (alpha-BuTX) with 1,2-cyclohexanedione resulted in the modification of only Arg-72 but arginine at position 36 or 72, as well as both were modified by reaction of the toxin with p-hydroxyphenylglyoxal. No derivative modified at Arg-25 was obtained, indicating that this residue may be located in the ... More
A fluorescence probe of acetylcholine receptor conformation and local anesthetic binding.
AuthorsSchimerlik M, Raftery MA
JournalBiochem Biophys Res Commun
PubMed ID1008877
A non-radioactive receptor assay for snake venom postsynaptic neurotoxins.
AuthorsStiles BG
JournalToxicon
PubMed ID1862522
'A non-radioactive assay was developed for detecting the binding of postsynaptic neurotoxins to acetylcholine receptor (AchR) from Torpedo californica. Enzyme linked immunosorbent assay (ELISA) wells coated with long or short chain neurotoxins specifically bound to purified AchR while crotamine or two different cardiotoxins did not. Bound receptor was detected by ... More
Activation and desensitization of Torpedo acetylcholine receptor: evidence for separate binding sites.
AuthorsDunn SM, Raftery MA
JournalProc Natl Acad Sci U S A
PubMed ID6960348
'The acetylcholine receptor from Torpedo californica was labeled by reaction with the fluorescent probe 4-[N-(iodoacetoxy)ethyl-N-methyl]amino-7-nitrobenz-2-oxa-1,3-diazole without apparent effect on its in vitro ligand binding and functional properties. Addition of acetylcholine or carbamoylcholine to the labeled-receptor preparations enhanced the fluorescence of the bound probe, and this effect was specific for agonists ... More
alpha-Latrotoxin releases calcium in frog motor nerve terminals.
AuthorsTsang CW, Elrick DB, Charlton MP
JournalJ Neurosci
PubMed ID11102474
'alpha-Latrotoxin (alpha-LTX) is a neurotoxin that accelerates spontaneous exocytosis independently of extracellular Ca(2+). Although alpha-LTX increases spontaneous transmitter release at synapses, the mechanism is unknown. We tested the hypothesis that alpha-LTX causes transmitter release by mobilizing intracellular Ca(2+) in frog motor nerve terminals. Transmitter release was measured electrophysiologically and with ... More
Substitution of Torpedo acetylcholine receptor alpha 1-subunit residues with snake alpha 1- and rat nerve alpha 3-subunit residues in recombinant fusion proteins: effect on alpha-bungarotoxin binding.
'A fusion protein consisting of the TrpE protein and residues 166-211 of the Torpedo acetylcholine receptor alpha 1 subunit was produced in Escherichia coli using a pATH10 expression vector. Residues in the Torpedo sequence were changed by means of oligonucleotide-directed mutagenesis to residues present in snake alpha 1 subunit and ... More
The cholinergic antagonist alpha-bungarotoxin also binds and blocks a subset of GABA receptors.
AuthorsMcCann CM, Bracamontes J, Steinbach JH, Sanes JR
JournalProc Natl Acad Sci U S A
PubMed ID16549768
'The polypeptide snake toxin alpha-bungarotoxin (BTX) has been used in hundreds of studies on the structure, function, and development of the neuromuscular junction because it binds tightly and specifically to the nicotinic acetylcholine receptors (nAChRs) at this synapse. We show here that BTX also binds to and blocks a subset ... More
Methyllycaconitine and (+)-anatoxin-a differentiate between nicotinic receptors in vertebrate and invertebrate nervous systems.
'Specific high-affinity binding sites for 125I-alpha-bungarotoxin and (-)-[3H]nicotine have been measured in rat brain and locust (Schistocerca gregaria) ganglia. The binding sites for 125I-alpha-bungarotoxin had similar Kd values of 1.5 x 10(-9) and 0.8 x 10(-9) M for rat and locust preparations, respectively; the corresponding values for the (-)-[3H]nicotine-binding site ... More
Chromatographic separation of the venom of Bungarus multicinctus and characterization of its components.
AuthorsLee CY, Chang SL, Kau ST, Luh SH
JournalJ Chromatogr
PubMed ID4403701
Nicotinic acetylcholine receptor superfamily of ligand-gated ion channels.
AuthorsStroud RM, McCarthy MP, Shuster M
JournalBiochemistry
PubMed ID1703009
Overview of toxins and drugs as tools to study excitable membrane ion channels: II. Transmitter-activated channels.
AuthorsNarahashi T
JournalMethods Enzymol
PubMed ID1382205
Interaction of [125I]-alpha-bungarotoxin with acetylcholine receptor from Torpedo californica.
AuthorsBlanchard SG, Quast U, Reed K, Lee T, Schimerlik MI, Vandlen R, Claudio T, Strader CD, Moore HP, Raftery MA
JournalBiochemistry
PubMed ID435451
Translational diffusion of acetylcholine receptor (monomeric and dimeric forms) of Torpedo marmorata reconstituted into phospholipid bilayers studied by fluorescence recovery after photobleaching.
AuthorsCriado M, Vaz WL, Barrantes FJ, Jovin TM
JournalBiochemistry
PubMed ID6897514
Fluorescent tetramethyl rhodamine derivatives of alpha-bungarotoxin: preparation, separation, and characterization.
AuthorsRavdin P, Axelrod D
JournalAnal Biochem
PubMed ID889094
Snake venoms: toolbox of the neurobiologist.
AuthorsMebs D
JournalEndeavour
PubMed ID2482807
Neurotoxins isolated from snake venom exhibit very specific actions on nervous structures. Their use in neurobiological research contributed to a better understanding of neuromuscular transmission and led to the isolation of a receptor, the nicotinic acetylcholine receptor. ... More
Delphinium alkaloids as inhibitors of alpha-bungarotoxin binding to rat and insect neural membranes.
AuthorsKukel CF, Jennings KR
JournalCan J Physiol Pharmacol
PubMed ID8012891
A series of C19-diterpenoid alkaloids purified from Delphinium were evaluated as inhibitors of alpha-bungarotoxin binding to rat and house fly neural membranes. In comparing these diterpenoid analogs, a wide range of inhibition potencies (IC50) were observed, with calculated IC50 values ranging six orders of magnitude. The most potent inhibitory alkaloids ... More
(1-Pyrene)sulfonyl azide: a fluorescent probe for measuring the transmembrane topology of acetylcholine receptor subunits.
AuthorsClarke J, Garcia-Borron JC, Martinez-Carrion M
JournalArch Biochem Biophys
PubMed ID3606117
(1-Pyrene)sulfonyl azide (PySA), a fluorescent, lipophilic photolabel, was used as a probe for the transmembrane topology of the acetylcholine receptor (AchR) subunits. Photolabeling of native, alkaline-extracted, and reconstituted AchR membrane preparations resulted in the labeling of all the AchR subunits. However the reconstituted AchR membrane preparations incorporated twice as much ... More
Methyllycaconitine: a selective probe for neuronal alpha-bungarotoxin binding sites.
AuthorsWard JM, Cockcroft VB, Lunt GG, Smillie FS, Wonnacott S
JournalFEBS Lett
PubMed ID2226787
The ability of methyllycaconitine (MLA) to inhibit the binding of [125I]alpha-bungarotoxin to rat brain membranes, frog and human muscle extracts and the human muscle cell line TE671 has been measured. MLA showed a markedly higher affinity for the rat brain site (Ki 1.4 x 10(-9) M) than for the muscle ... More
Neurotoxins: overview of an emerging research technology.
AuthorsAdams ME, Olivera BM
JournalTrends Neurosci
PubMed ID7517594
Neurotoxins have highly specific actions on molecular targets, and thus offer an effective means of characterizing the growing number of identified ion channels and receptors in the nervous system. This article and the Neurotoxins Supplement accompanying this issue of TINS provide a convenient reference source to facilitate the use of ... More
The role of an invariant tryptophan residue in alpha-bungarotoxin and cobrotoxin. Investigation of active derivatives with the invariant tryptophan replaced by kynurenine.
AuthorsChang CC, Kawata Y, Sakiyama F, Hayashi K
JournalEur J Biochem
PubMed ID2226470
Ozone oxidation converted the single, invariant, tryptophan residue to N2-formylkynurenine in alpha-bungarotoxin and cobrotoxin. Upon this modification, the lethal toxicity was significantly reduced in cobrotoxin but mostly retained in alpha-bungarotoxin. Each neurotoxin containing kynurenine instead of tryptophan retained the same antigenicity as the native toxin. Fluorescence and CD spectroscopy revealed ... More
Requirement for cholinergic synaptic transmission in the propagation of spontaneous retinal waves.
AuthorsFeller MB, Wellis DP, Stellwagen D, Werblin FS, Shatz CJ
JournalScience
PubMed ID8638165
Highly correlated neural activity in the form of spontaneous waves of action potentials is present in the developing retina weeks before vision. Optical imaging revealed that these waves consist of spatially restricted domains of activity that form a mosaic pattern over the entire retinal ganglion cell layer. Whole-cell recordings indicate ... More
Nicotine-induced intracellular calcium changes are not antagonized by alpha-bungarotoxin in adrenal medullary cells.
AuthorsAfar R, Trifaró JM, Quik M
JournalBrain Res
PubMed ID8019838
The snake toxin alpha-bungarotoxin distinguishes between neuronal nicotinic receptor subtypes. In chick ciliary ganglion neurons, activation of alpha-bungarotoxin-sensitive nicotinic receptors has been proposed to produce elevations in intracellular calcium levels. In the present study we show that prolonged treatment with alpha-bungarotoxin did not affect the nicotine-evoked calcium response in suspended ... More
Affinity-directed cross-linking of membrane-bound acetylcholine receptor polypeptides with photolabile alpha-bungarotoxin derivatives.
AuthorsWitzemann V, Muchmore D, Raftery MA
JournalBiochemistry
PubMed ID518854
Photolabile derivatives of [125I]-alpha-bungarotoxin that retain specific binding to Torpedo californica acetylcholine receptor have been utilized as structural probes of the receptor complex of polypeptide components in its membrane-associated form. The derivatized toxins contained aryl azide side chains poised to form covalent cross-links to both associated and adjacent polypeptides following ... More
Synaptic regulation of glial protein expression in vivo.
We investigated signaling between individual nerve terminals and perisynaptic Schwann cells, the teloglial cells that cover neuromuscular junctions. When deprived of neuronal activity in vivo, either by motor nerve transection or tetrodotoxin injection, perisynaptic Schwann cells rapidly up-regulated glial fibrillary acidic protein. Addition of transcription or translation inhibitors to excised ... More
Intracellular calcium homeostasis in a human neuroblastoma cell line: modulation by depolarization, cholinergic receptors, and alpha-latrotoxin.
AuthorsSher E, Gotti C, Pandiella A, Madeddu L, Clementi F
JournalJ Neurochem
PubMed ID2453611
Intracellular calcium homeostasis and its modulation by different agents was studied in control and differentiated IMR32 human neuroblastoma cells by using the Ca2+-sensitive fluorescent dye quin2. The results obtained demonstrate the existence in IMR32 cells of (a) voltage-dependent, verapamil sensitive, Ca2+ channels, which are expressed before differentiation; (b) muscarinic receptors ... More
A role for the nicotinic alpha-bungarotoxin receptor in neurite outgrowth in PC12 cells.
AuthorsChan J, Quik M
JournalNeuroscience
PubMed ID8247271
The addition of nicotine decreased neuritic outgrowth in PC12 cells in culture. This effect occurs as early as one day after addition of nicotine to the culture medium in a concentration-dependent manner. The nicotine-induced decline in neurite outgrowth was prevented by d-tubocurarine (10(-4) M) indicating that the effect was mediated ... More
Blockade of nicotinic currents in hippocampal neurons defines methyllycaconitine as a potent and specific receptor antagonist.
AuthorsAlkondon M, Pereira EF, Wonnacott S, Albuquerque EX
JournalMol Pharmacol
PubMed ID1569927
Methyllycaconitine, a toxin isolated from the seeds of Delphinium brownii, inhibited acetylcholine- and anatoxin-induced whole-cell currents in cultured fetal rat hippocampal neurons, at picomolar concentrations. This antagonism was specific, concentration dependent, reversible, and voltage independent. Furthermore, methyllycaconitine inhibited 125I-alpha-bungarotoxin binding to adult rat hippocampal membranes, protected against the alpha-bungarotoxin-induced pseudoirreversible ... More
Direct spectroscopic studies of cation translocation by Torpedo acetylcholine receptor on a time scale of physiological relevance.
AuthorsMoore HP, Raftery MA
JournalProc Natl Acad Sci U S A
PubMed ID6933499
The kinetics of carbamoylcholine-mediated cation transport across the membrane of vesicles containing acetylcholine receptor have been measured on the physiologically relevant time scale of a few milliseconds. The stopped-flow spectroscopic approach utilizes thallium(I) as the cation transported into sealed vesicles containing a water-soluble fluorophore. Upon entry of thallium(I), fluorescence quenching ... More
Kinetics of interaction of N epsilon-fluorescein isothiocyanate-lysine-23-cobra alpha-toxin with the acetylcholine receptor.
AuthorsCheung AT, Johnson DA, Taylor P
JournalBiophys J
PubMed ID6696970
We have studied the kinetics of N epsilon-fluorescein isothiocyanate-lysine-23 cobra alpha-toxin (FITC-toxin) binding to the membrane-associated acetylcholine receptor from the Torpedo californica electric organ. The fluorescent toxin not only enabled us to monitor the binding reaction continuously but also to examine simultaneously the enhancement of ligand fluorescence and the increase ... More
Characterization of the binding of alpha-bungarotoxin to bacterially expressed cholinergic binding sites.
AuthorsAronheim A, Eshel Y, Mosckovitz R, Gershoni JM
JournalJ Biol Chem
PubMed ID3290216
Bacterially expressed cDNA fragments of the alpha-subunit of the nicotinic acetylcholine receptor previously have been shown to bind alpha-bungarotoxin (Gershoni, J. M. (1987) Proc. Natl. Acad. Sci. U. S. A. 84, 4318-4321). Here, a novel system has been developed in which totally synthetic alpha-bungarotoxin binding sites are expressed in Escherichia ... More
Status of tryptophan residue in cobrotoxin and alpha-bungarotoxin.
AuthorsChang LS, Kuo KW, Chang CC
JournalBiochem Mol Biol Int
PubMed ID8485461
Acrylamide quenching studies indicated that the exposure degree of the Trp residue in cobrotoxin was higher than that in alpha-bungarotoxin. The Trp residue of cobrotoxin was in a positively charged environment as revealed by iodide quenching, while the Trp residue of alpha-bungarotoxin was not accessible for iodide. Analysis of hydrophilicity ... More
Rotational molecular dynamics of the membrane-bound acetylcholine receptor revealed by phosphorescence spectroscopy.
AuthorsBartholdi M, Barrantes FJ, Jovin TM
JournalEur J Biochem
PubMed ID6172277
The rotational mobility of the membrane-bound acetylcholine receptor from Torpedo marmorata has been studied by phosphorescence anisotropy techniques using eosin-5'-isothiocyanate and eosin-5'-iodoacetamide derivatives of the alpha-neurotoxin of Bungarus multicinctus (alpha-bungarotoxin). Normal membranes, those depleted of nonreceptor peripheral peptides by alkaline treatment, as well as membranes subjected to various chemical modifications ... More
Hippocampal synaptic transmission enhanced by low concentrations of nicotine.
AuthorsGray R, Rajan AS, Radcliffe KA, Yakehiro M, Dani JA
JournalNature
PubMed ID8878480
Nicotine obtained from tobacco can improve learning and memory on various tasks and has been linked to arousal, attention, rapid information processing, working memory, and long-term memories that can cause craving years after someone has stopped smoking. One likely target for these effects is the hippocampus, a centre for learning ... More
Acetylcholinesterase dynamics at the neuromuscular junction of live animals.
AuthorsKrejci E, Martinez-Pena y Valenzuela I, Ameziane R, Akaaboune M
JournalJ Biol Chem
PubMed ID16455662
At cholinergic synapses, acetylcholinesterase (AChE) is critical for ensuring normal synaptic transmission. However, little is known about how this enzyme is maintained and regulated in vivo. In this work, we demonstrate that the dissociation of fluorescently-tagged fasciculin 2 (a specific and selective peptide inhibitor of AChE) from AChE is extremely ... More
Neuronal acetylcholine receptors in Drosophila: the ARD protein is a component of a high-affinity alpha-bungarotoxin binding complex.
AuthorsSchloss P, Hermans-Borgmeyer I, Betz H, Gundelfinger ED
JournalEMBO J
PubMed ID3141150
The ard gene of Drosophila melanogaster encodes a structural homologue of vertebrate nicotinic acetylcholine receptors (AChR) and is expressed exclusively in nervous tissue. To study the nature of the ARD protein, antibodies were raised against fusion constructs containing two regions of this polypeptide. One segment is putatively extracellular (amino acids ... More
Chemical transduction with fluorescent lipid membranes using selective interactions of acetylcholine receptor with agonist/antagonist and acetylcholine with substrate.
AuthorsKrull UJ, Brennan JD, Brown RS, Hosein S, Hougham BD, Vandenberg ET
JournalAnalyst
PubMed ID2321768
Alterations in the physical structure of vesicles and monolayers of phospholipids and soybean lecithin were monitored by measurement on the average fluorescence intensity changes from N-(7-nitrobenz-2-oxa-1,3-diazol-4-yl)dipalmitoyl-L-a-phosphatidyl ethanolamine (NBD-PE) located in the lipid matrices. This probe was intimately dispersed at a concentration of 1-2 mol-% in lipid membranes and had an ... More
alpha-Bungarotoxin immobilized and oriented on a lipid bilayer vesicle surface.
AuthorsBabbitt B, Huang L
JournalBiochemistry
PubMed ID3994962
We have developed a new method to assess the binding site on alpha-bungarotoxin (alpha-BGT) for the acetylcholine receptor. It involves the covalent attachment of a palmitic acid chain to the toxin molecule, generating monopalmitoyl-alpha-bungarotoxin (PBGT) which is then immobilized on the surface of a lipid vesicle by a process of ... More
Localization of L-type Ca2+ channels at perisynaptic glial cells of the frog neuromuscular junction.
AuthorsRobitaille R, Bourque MJ, Vandaele S
JournalJ Neurosci
PubMed ID8613781
The presence of L-type Ca2+ channels at the frog neuromuscular junction (nmj) was studied by monitoring changes in intracellular Ca2+ evoked in presynaptic terminals and perisynaptic Schwann cells (PSCs) and by studying the distribution of Ca2+ channels using a monoclonal antibody directed against the alpha 2/delta subunit of L channels. ... More
Laminin-1 redistributes postsynaptic proteins and requires rapsyn, tyrosine phosphorylation, and Src and Fyn to stably cluster acetylcholine receptors.
AuthorsMarangi PA, Wieland ST, Fuhrer C
JournalJ Cell Biol
PubMed ID12034776
Clustering of acetylcholine receptors (AChRs) is a critical step in neuromuscular synaptogenesis, and is induced by agrin and laminin which are thought to act through different signaling mechanisms. We addressed whether laminin redistributes postsynaptic proteins and requires key elements of the agrin signaling pathway to cause AChR aggregation. In myotubes, ... More
Ultrastructural distribution of alpha-bungarotoxin binding sites in the suprachiasmatic nucleus of the rat hypothalamus.
AuthorsMiller MM, Billiar RB, Beaudet A
JournalCell Tissue Res
PubMed ID3652154
The distribution of (125I) alpha bungarotoxin (alpha-BTX) binding sites in the suprachiasmatic nucleus (SCN) of the adult female rat was examined by electron-microscopic autoradiography. The ultrastructural distribution of silver grains was analysed by line source, direct point count, and 50% probability circle methods. Real grain distribution was significantly different from ... More
1-Pyrene-butyrylcholine: a fluorescent probe for the cholinergic system.
The action of 1-pyrene-butyrylcholine, a new cholinergic fluorescent probe, has been studied at the cellular level using electrophysiological and fluorescence techniques. The spectroscopic properties of the probe were found to be similar to those pf pyrene-butyric acid, the excited-state lifetime in air-saturated aqueous solutions being 92 nsec. At micromolar concentrations ... More
Reductive methylation as a tool for the identification of the amino groups in alpha-bungarotoxin interacting with nicotinic acetylcholine receptor.
AuthorsGarcia-Borron JC, Bieber AL, Martinez-Carrion M
JournalBiochemistry
PubMed ID3663591
alpha-Bungarotoxin (alpha Bgt) is a postsynaptic neurotoxin which blocks cholinergic transmission at the neuromuscular junction by binding tightly to the acetylcholine receptor (AcChR). The number of methylation sites in alpha Bgt has been shown to decrease significantly upon binding of the toxin to the AcChR [Soler, G., Farach, M. C., ... More
Perisynaptic surface distribution of multiple classes of nicotinic acetylcholine receptors on neurons in the chicken ciliary ganglion.
AuthorsHorch HL, Sargent PB
JournalJ Neurosci
PubMed ID8613719
Most nicotinic ACh receptors (AChRs) studied to date in the embryonic chicken ciliary ganglion are recognized either by monoclonal antibody (mAb) 35 or by alpha-bungarotoxin (alpha-Bgt). Previous studies found that mAb 35-AChRs are found at both synaptic and extrasynaptic sites, while alpha-Bgt-AChRs are found exclusively at extrasynaptic sites. To gain ... More
Purinergic receptors and their activation by endogenous purines at perisynaptic glial cells of the frog neuromuscular junction.
AuthorsRobitaille R
JournalJ Neurosci
PubMed ID7472466
Glial cells are closely associated with synapses and are modulated by neurotransmitters released during synaptic transmission. At many synapses, ATP is released during synaptic transmission and is involved in cell-cell signaling. Since glial cells have purinoceptors, it is possible that ATP mediates synaptic neuron-glia signaling. This work aims at determining ... More
Synaptosomal binding of 125I-labelled daboiatoxin, a new PLA2 neurotoxin from the venom of Daboia russelli siamensis.
AuthorsMaung-Maung-Thwin Gopalakrishnakone P, Yuen R, Tan CH
JournalToxicon
PubMed ID8711753
Daboiatoxin (DbTx), the PLA2 neurotoxin from Daboia russelli siamensis venom, was shown to bind specifically and saturably to rat cerebrocortical synaptosomes and synaptic membrane fragments. Two families of binding sites were detected by equilibrium binding analysis in the presence and absence of Ca2+. Scatchard analysis of biphasic plateaus revealed Kdl ... More
Probing local secondary structure by fluorescence: time-resolved and circular dichroism studies of highly purified neurotoxins.
AuthorsDahms TE, Szabo AG
JournalBiophys J
PubMed ID8527671
The relationship between beta-sheet secondary structure and intrinsic tryptophan fluorescence parameters of erabutoxin b, alpha-cobratoxin, and alpha-bungarotoxin were examined. Nuclear magnetic resonance and x-ray crystallography have shown that these neurotoxins have comparable beta-sheet, beta-turn, and random coil secondary structures. Each toxin contains a single tryptophan (Trp) residue within its beta-sheet. ... More
Novel biotinylated phenylarsonous acids as bifunctional reagents for spatially close thiols: studies on reduced antibodies and the agonist binding site of reduced Torpedo nicotinic receptors.
AuthorsMoaddel R, Sharma A, Huseni T, Jones GS, Hanson RN, Loring RH
JournalBioconjug Chem
PubMed ID10411461
We synthesized three novel organoarsenicals as prototype bifunctional reagents for spatially close thiols, N-(4-arsenosophenyl) hexahydro-2-oxo-(3aS,4S,6aR)-1H-thieno[3, 4-d]imidazole-4-pentamide (1), 2-[4-[(4-arsenosophenyl)amino]-1, 4-dioxobutyl] hydrazide, (3aS,4S,6aR)-hexahydro-2-oxo- 1H-thieno[3, 4-d] imidazole-4-pentanoic acid (2), and [4-[[12-[[5-[(3aS,4S, 6aR)-hexahydro-2-oxo-1H-thieno[3, 4-d]imidazol-4-yl]-1-oxopentyl]amino]-1-oxododecyl]amino]phe nyl]-arso nous acid (3) containing both biotin and arsenic with intervening varying length spacers extending from 2 to 15 A ... More
Identification of a brain acetylcholine receptor alpha subunit able to bind alpha-bungarotoxin.
AuthorsMcLane KE, Wu XD, Conti-Tronconi BM
JournalJ Biol Chem
PubMed ID2351675
Peptides corresponding to sequence segments homologous to an alpha-bungarotoxin (alpha-BGT) binding region on the alpha subunit of the Torpedo nicotinic cholinergic receptor (nAChR) were synthesized for each identified nAChR alpha subunit of the rat nervous system (alpha 1, which is expressed in muscle, and alpha 2, alpha 3, alpha 4, ... More
Interaction of monoclonal antibodies with alpha-bungarotoxin and (-)-nicotine binding sites in goldfish brain. Identification of putative nicotinic acetylcholine receptor subtypes.
AuthorsHenley JM, Lindstrom JM, Oswald RE
JournalJ Biol Chem
PubMed ID3290212
Monoclonal antibodies raised against the nicotinic acetylcholine receptor of Electrophorus electricus electroplaque have been used as probes to characterize putative nicotinic acetylcholine receptors in goldfish brain. One monoclonal antibody (mAb), mAb 47, recognized a protein which binds both (-)-[3H]nicotine and 125I-alpha-bungarotoxin with high affinity. Another monoclonal antibody (mAb 172) recognized ... More
Differential regulation of neuronal nicotinic ACh receptor subunit genes in cultured neonatal rat sympathetic neurons: specific induction of alpha 7 by membrane depolarization through a Ca2+/calmodulin-dependent kinase pathway.
AuthorsDe Koninck P, Cooper E
JournalJ Neurosci
PubMed ID8613734
We have examined the regulation of neuronal nicotinic ACh receptor (nAChR) genes and ACh-evoked currents by neonatal rat sympathetic neurons developing in culture. These neurons contain 5 nAChR transcripts: alpha 3, alpha 5, alpha 7, beta 2, and beta 4. When developing in culture, the neurons express 4 of these ... More
Retinotectal circuitry of larval zebrafish is adapted to detection and pursuit of prey.
Authors
JournalElife
PubMed ID33044168
Specialized neurons in the right habenula mediate response to aversive olfactory cues.
Authors
JournalElife
PubMed ID34878403
Repairing Volumetric Muscle Loss in the Ovine Peroneus Tertius Following a 3-Month Recovery.
Authors
JournalTissue Eng Part A
PubMed ID32013753
Neuronal Architecture of a Visual Center that Processes Optic Flow.
Authors
JournalNeuron
PubMed ID31147153
From Whole-Brain Data to Functional Circuit Models: The Zebrafish Optomotor Response.
Authors
JournalCell
PubMed ID27814522
Bypassing Glutamic Acid Decarboxylase 1 (Gad1) Induced Craniofacial Defects with a Photoactivatable Translation Blocker Morpholino.
Authors
JournalACS Chem Neurosci
PubMed ID30200754
Liprin-α-1 is a novel component of the murine neuromuscular junction and is involved in the organization of the postsynaptic machinery.
Authors
JournalSci Rep
PubMed ID28831123
Drebrin Regulates Acetylcholine Receptor Clustering and Organization of Microtubules at the Postsynaptic Machinery.
Authors
JournalInt J Mol Sci
PubMed ID34502296
A novel bungarotoxin binding site-tagged construct reveals MAPK-dependent Kv4.2 trafficking.
Authors
JournalMol Cell Neurosci
PubMed ID31212013
Site-directed MT1-MMP trafficking and surface insertion regulate AChR clustering and remodeling at developing NMJs.