Journal ArticleDOI
Iodide, thiocyanate and cyanide ions as capturing reagents in one-step copper-thiocholine method for cytochemical localization of cholinesterase activity.
Miro Brzin,Simona Pucihar +1 more
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Better-quality localization in the presence of either CN− or CNS− suggests that they, and particularly CN−, should be substituted for I− in the one-step Cu-ThCh method for the cytochemistry of cholinesterases.Abstract:
The necessity of the presence of iodide in Cu-ThCh reaction was investigated by following the precipitate formation "in vitro" and by evaluating the ultrastructural localization of the precipitate in sympathetic ganglion cells of the frog and in the end-plate regions of the rat diaphragm. It was found that thiocyanate or cyanide is the only anion that can be substituted for iodide as the capturing agent in precipitation. The optimal concentration in the preincubation and incubation media of any one of the three anions is from 2 to 5 mM. At a concentration below 1 mM precipitation "in vitro" is considerably delayed as a result of which in electron microscopy diffusion artefacts appear in tissue sections. The unconverted primary precipitate obtained in the presence of iodide had been used for ultrastructural localization of ChE activity and now this use has been extended to precipitates obtained in the presence of CN- or CNS-. Better-quality localization in the presence of either one of the latter anions suggests that they, and particularly CN-, should be substituted for I- in the one-step Cu-ThCh method for the cytochemistry of cholinesterases.read more
Citations
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Journal ArticleDOI
The Molecular Forms of Cholinesterase and Acetylcholinesterase in Vertebrates
J Massoulié,S Bon +1 more
TL;DR: The mechanism of cholinergic neurotransmission requires the rapid inac tivation of acetylcholine, which exists in all classes of vertebrates and is characterized in horse serum by Stedman et al (1932), who called it choli nesterase.
Journal ArticleDOI
Differentiation of glial cells and motor neurons during the formation of neuromuscular junctions in cocultures of rat spinal cord explant and human muscle.
Tomaz Mars,Kevin J. Yu,Xue-Ming Tang,Armand F. Miranda,Zoran Grubič,Franca Cambi,Michael P. King +6 more
TL;DR: The hypothesis that glial cells are a major contributor to maturity of the neuromuscular junction formed in vitro in spinal cord explant–muscle cocultures is supported.
Journal ArticleDOI
Myoblast fusion and innervation with rat motor nerve alter distribution of acetylcholinesterase and its mRNA in cultures of human muscle
TL;DR: Coincidence of mRNA down-regulation and synthesis of stable basal lamina-bound AChE suggests coordinated adaptation, so that sufficient enzyme may be derived from low message levels, also supporting an earlier proposal that transcription proceeds intermittently.
Journal ArticleDOI
Origin of acetylcholinesterase in the neuromuscular junction formed in the in vitro innervated human muscle.
TL;DR: This is the first report demonstrating that both motor neuron and myotube contribute synaptic AChE under conditions where they interact with each other in the formation of an intact and functional NMJ.
Journal ArticleDOI
Attachment of acetylcholinesterase to structures of the motor endplate.
Janez Sketelj,Miro Brzin +1 more
TL;DR: The kinetics of AChE solubilization from intact motor endplates of mouse diaphragm, by collagenase, papain and hyaluronidase was studied in parallel with the ultrastructural localization of A ChE in treated neuromuscular junctions.
References
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Journal ArticleDOI
A histochemical method for localizing cholinesterase activity.
TL;DR: A histochemical method is presented for localizing ChE activity by incubating tissue sections in a medium containing acetylthiocholine, copper glycinate and copper thiocholine with results obtained with several tissues containing specific ChE.
Journal ArticleDOI
The distribution of cholinesterase in cholinergic neurons demonstrated with the electron microscope.
P. R. Lewis,C. C. D.Shute +1 more
TL;DR: The thiocholine technique for cholinesterase has been successfully adapted for the demonstration of enzyme distribution with the electron microscope and the close correlation between the electron-microscopic results and evidence from physiological and biochemical investigations is discussed.
Journal ArticleDOI
ACETYLCHOLINESTERASE IN FROG SYMPATHETIC AND DORSAL ROOT GANGLIA : A Study by Electron Microscope Cytochemistry and Microgasometric Analysis with the Magnetic Diver.
TL;DR: The localization and chemical determination of acetylcholin esterase in the frog sympathetic and dorsal root ganglia were studied by a combination of the methods of electron microscopy, histochemistry, and microgasometric analysis with the magnetic diver to show the effects of fixation on enzyme activity.
Journal ArticleDOI
A Modification of the Thiocholine Method for the Determination of Cholinesterase. II. Histochemical Application.
TL;DR: With the modified thiocholine method, stellate ganglion, striated muscle and stomach wall have been stained in order to produce confirmation of the biochemical controls, not with the purpose of studying the morphology in detail.
Journal ArticleDOI
On the chemical basis of thiocholine methods for demonstration of acetylcholinesterase activities
TL;DR: The reaction mechanism of “Koelle's thiocholine method” and "Karnovsky's ferricyanide method" for the localization of acetylcholinesterase activity have been studied and some modifications for these two methods have been proposed.