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Query: EC:3.1.1.7 (
acetylcholinesterase
)
28,390
document(s) hit in 31,850,051 MEDLINE articles (0.00 seconds)
By use of different inhibitors as well as atomic absorption spectrophotometry it has been shown that the haemolymph-
acetylcholinesterase
(E. c. 3.1.1.7) of the sea mussel Mytilus edulis is a metalloprotein containing 2,95 Fe2+-ions per subunit. All inhibitors used (1,10-phenanthroline, salicylic
aldehyde
, 2,2'-dipyridyl, 8-hydroxyquinoline) showed a non-competitive inhibition, which was not pH-dependent. Some divalent cations caused a marked increase of the enzyme activity, some heavy metals inhibited the enzyme almost completely; monovalent inorganic cations did not influence the enzyme at all. Besides NaF and Na2SiF6, which showed a non-competitive inhibition comparable to the inhibition observed with the chelating agents, and NaN3, whose mode of action was not identifiable, no inhibition by different mono- and divalent inorganic anions was to be observed. Ammonium ions caused no enzyme inhibition, but length the inhibition power of substituted ammonium ions increased with an increasing C-chain. The influence of some organic solvents on the enzyme activity is demonstrated.
...
PMID:Influence of ions and chelating agents on the haemolymphacetylcholinesterase of Mytilus edulis. 15 22
A simple and rapid method for the estimation of the hydrolysis of succinyl choline by serum
cholinesterase
variants is described. Succinyl choline, as substrate for the enzyme assay, has many advantages over other substrates (acetyl choline, benzoyl choline and butyryl choline) which have no clinical application. Choline, the hydrolytic product of succinyl choline, is oxidized to betaine
aldehyde
by choline oxidase (EC 1.1.99.1), a rat liver mitochondrial preparation; this is coupled to the reduction of cytochrome c which is measured at 550 nm. Fifty normal sera (UU), 17 heterozygous (UA) and 8 atypical (AA) were tested with this method, and on the basis of resistance to dibucaine (Cinchocain; Kalow, W. & Genest, K. (1957) Canad. J. Biochem. Physiol. 35, 339-346) inhibition, three distinct groups could be established using succinyl choline as substrate. These results are comparable with the standard optical method of Kalow & Genest (cf. above) using benzoyl choline as substrate.
...
PMID:[A spectrophotometric method for the determination of serum cholinesterase variants with succinyl choline as substrate (author's transl)]. 16 94
In order to carry out ultrastructural immunocytochemical localization of
acetylcholinesterase
at cholinergic synapses, ultrathin frozen sections of
aldehyde
-fixed electric organs of Electrophorus and Torpedo were obtained by a modified ultracryotome technique. Dry frozen sections, picked up with rabbit serum and negatively stained, revealed, in the postsynaptic regions of the neuro-electroplaque junction, the presence of numerous rod-like or pin-headed protrusions (30 X 40 A) attached perpendicularly at 60 A intervals to the hydrophobic lamina of the plasma membrane, forming a comb-like structure oriented toward the synaptic cleft. A possible correlation is suggested between this comb-like structure and the 'acetylcholine receptors' observed by other authors with high resolution electron microscopy either after classical preparative techniques or after freeze-etching, of the postsynaptic membrane of vertebrae cholinergic synapses.
...
PMID:Ultracryotomy of nerve-electroplaque synapses for immunocytochemistry. 35 Nov 46
Liquor contacting peptidergic neurons (LCPNs) in the preoptic nucleus of the Japanese eel (Anguilla japonica), are investigated submacroscopically, light microscopically, electron microscopically (transmission and scanning) and histochemically. LCPNs appear in 8--13 per cent of all neurons constituting the preoptic nucleus and their cytoplasm contains many secretory granules stained by
aldehyde
-thionin or chrome hematoxylin. LCPNs have an epithelial cell-like polarity and their cytoplasmic organella shift to the supranuclear region. LCPNs are classified into three types (A, B, C) according to the liquor contacting portion of the cell: Granular type A neuron (40--50 x 40--50 microns 2), the cell of which is in contact with the cerebrospinal fluid (CSF), is the most common type and distributed in the ventral portion of the preoptic nucleus; this neuron is not connected with the neighboring ependymal cells by tight junctions. Bipolar type B neuron (60 x 30 micron 2), contacts the CSF with the tip of it cell process and is scattered throughout the preoptic nucleus; the cell is connected with the surrounding ependymal cells by tight junction. Bipolar type C neuron (60 x 30 micron 2) possesses a cell process protruded into the third ventricle and is distributed in the dorsal portion of the preoptic nucleus; this also is connected with the adjacent ependymal cells by tight junction. Regardless of type, all LCPNs exhibit a positive
acetylcholinesterase
and a negative ATPase reaction. Numerous fluorescent varicosities of monoaminergic nerve terminals are closely associated with the cell bodies of the LCPN. LCPNs are likely regulated by monoaminergic fibers.
...
PMID:Histological and cytological studies on the liquor contacting peptidergic neurons in the preoptic nucleus of the Japanese eel (Anguilla japonica). 53 83
Light microscopic observations using Nomarski optics on the
aldehyde
-fixed hypothalamus of normal adult cats, monkeys and rabbits revealed the presence of cells in the supraoptic, paraventricular and periventricular nuclei which possessed yellow birefringent inclusions. Immunogold labelling showed that in each species the cells displayed oxytocin-like immunoreactivity, both in electron-dense inclusions within some (but not all) cisterns of rough endoplasmic reticulum and in secretory granules. The cells in cats and rabbits were in all respects indistinguishable from the homologous 'birefringent' cells previously described in rats, but in monkeys, cells frequently contained additional inclusions in cisterns of rough endoplasmic reticulum which did not display oxytocin or vasopressin-like immunoreactivity, even after trypsin, pepsin or chymotrypsin treatment of sections. Observations on cats and rabbits using fluorescence microscopy revealed that the birefringent cells possessed bright autofluorescence which facilitated the identification of more cells than were seen using Nomarski optics alone. Autofluorescence was abolished when sections were mounted in glycerol, or when exposed to light for protracted periods of time. Attempts to label for monoamines in these cells were not successful, suggesting that the fluorescence is not due to
aldehyde
-induced amine fluorescence. It is not clear why neuropeptides are retained in some rough endoplasmic reticulum cisterns. It is possible that these birefringent cells contain a peptide, or peptides, which are abnormal in some manner, or which may be other members of the oxytocin gene family. Alternatively, the processing of neuropeptides to permit their export to the Golgi apparatus may be deficient. Acetylcholinesterase (AChE) histochemistry revealed that, unlike other oxytocin neurons, cells with intracellular accretions lacked detectable acetyl
cholinesterase
. As AChE is a known peptidase, it may be involved in regulating peptide export from the rough endoplasmic reticulum.
...
PMID:Neuropeptide accretions in the endoplasmic reticulum of oxytocinergic neurons in cats, monkeys and rabbits: a widespread phenomenon. 129 66
Although a well-developed plexus of nerves and ganglia is known to be present in the wall of the gallbladder, little has previously been learned about the function or organization of this innervation. The current study was undertaken in order to evaluate the hypothesis that the ganglionated plexus of the gallbladder is analogous to elements of the enteric nervous system (ENS). The ganglionated plexus of the gallbladder was found to resemble closely the submucosal plexus of the small intestine in its organization into two irregular anastomosing and interwoven networks of ganglia, in the numbers of neurons per ganglion, and in the manifestation of histochemically demonstrable
acetylcholinesterase
activity in virtually all ganglion cells. In common with enteric ganglia, laminin immunoreactivity was observed to be excluded from the interiors of gallbladder ganglia, which were surrounded by a periganglionic laminin-immunoreactive sheath. As in the submucosal plexus, intrinsic substance P-, vasoactive intestinal polypeptide (VIP)-, and neuropeptide Y (NPY)-immunoreactive neurons were seen in the ganglionated plexus of the gallbladder. Extrinsic nerves in the gallbladder that degenerated following chemical sympathectomy with 6-hydroxydopamine (6-OHDA), and which contained NPY, tyrosine hydroxylase (TH), and dopamine-beta-hydroxylase (DBH) immunoreactivities, formed a perivascular plexus closely associated with blood vessels. Endogenous catecholamines could also be demonstrated in these perivascular nerves by
aldehyde
-induced histofluorescence. In addition to perivascular nerves, paravascular nerve bundles were observed that were loosely associated with vessels, did not degenerate following administration of 6-OHDA, and contained NPY immunoreactivity. Other paravascular nerves, probably visceral sensory axons, coexpressed substance P and calcitonin-gene-related peptide (CGRP) immunoreactivities. The ganglionated plexus of the gallbladder resembled enteric ganglia in having intrinsic 5-hydroxytryptamine (5-HT)-immunoreactive cells and highly varicose nerve fibers. The 5-HT-immunoreactive gallbladder axons were, like those of the gut, resistant to 6-OHDA, and separate from fibers that expressed TH immunoreactivity. Differences between the ganglionated plexus of the gallbladder and enteric ganglia of the small intestine included in the gallbladder are 1) the presence of TH-immunoreactive cells that contain an endogenous catecholamine, but not DBH; 2) DBH-immunoreactive neurons, some of which coexpress substance P immunoreactivity, but which contain neither a catecholamine nor TH immunoreactivity; 3) an apparent absence of CGRP-immunoreactive cell bodies.(ABSTRACT TRUNCATED AT 400 WORDS)
...
PMID:Structure, afferent innervation, and transmitter content of ganglia of the guinea pig gallbladder: relationship to the enteric nervous system. 256 71
Onchidal has been identified as the major lipid-soluble component of the defensive secretion of the mollusc Onchidella binneyi, and it has been proposed as the compound responsible for the chemical protection of Onchidella [Bioorg. Chem. 7:125-131 (1978)]. In support of this hypothesis, we now report that onchidal can be found in several different species of Onchidella and that it is toxic to fish. Because onchidal is an acetate ester similar to acetylcholine, its ability to interact with nicotinic acetylcholine receptors and
acetylcholinesterase
was investigated. Although onchidal did not prevent the binding of 125I-alpha-bungarotoxin to nicotinic acetylcholine receptors, it inhibited
acetylcholinesterase
in a progressive, apparently irreversible, manner. The apparent affinity of onchidal for the initial reversible binding to
acetylcholinesterase
(Kd) was approximately 300 microM, and the apparent rate constant for the subsequent irreversible inhibition of enzyme activity (kintact) was approximately 0.1 min-1. Onchidal was a substrate for
acetylcholinesterase
, and approximately 3250 mol of onchidal were hydrolyzed/mol of enzyme irreversibly inhibited. The calculated kcat for onchidal was 325 min-1. Irreversible inhibition resulted from either onchidal itself or a reactive intermediate in the enzyme-catalyzed hydrolysis of onchidal, rather than from the hydrolysis products of onchidal. Irreversible inhibition of enzyme activity was prevented by coincubation with reversible agents that either sterically block (edrophonium and decamethonium) or allosterically modify (propidium) the acetylcholine binding site. Enzyme activity was not regenerated by incubation with oxime reactivators; therefore, the mechanism of irreversible inhibition does not appear to involve acylation of the active site serine. Because onchidal contains a potentially reactive alpha,beta-unsaturated
aldehyde
, irreversible inhibition of
acetylcholinesterase
may result from formation of a novel covalent bond between the toxin and the enzyme. Thus, this novel toxin could potentially be exploited in the design of a new class of anticholinesterase insecticides and in the identification of amino acids that contribute to the binding and hydrolysis of acetylcholine.
...
PMID:Onchidal: a naturally occurring irreversible inhibitor of acetylcholinesterase with a novel mechanism of action. 277 21
Immunocytochemical evidence is presented for the existence of choline acetyltransferase (ChoAcTase), cysteine sulfinic acid decarboxylase (CSADCase), tyrosine hydroxylase (TyrOHase), and glutamic acid decarboxylase (GluDCase) in large motor neurons of the hypoglossal nucleus and the spinal cord and in nerve terminals of motor end plates in tongue and skeletal muscle of five mammalian species, including man. These enzymes, which are responsible for the synthesis of acetylcholine (AcCho), taurine, dopamine, and gamma-aminobutyrate (GABA), respectively, were detected by immunocytochemical studies with monoclonal or polyclonal antibodies raised against the enzymes. Electron microscopy of the neuromuscular junctions showed that the immunoreactivity in each case was confined to the cytoplasmic matrix of presynaptic nerve terminals. Immunoreactivity obtained for each enzyme antibody varied with the species. It was highest in fresh, unfixed muscle and lowest in
aldehyde
-fixed specimens. Negative controls were obtained with preimmune sera and antisera preabsorbed with pure ChoAcTase, CSADCase, or GluDCase antigen. Double-labeling studies with ChoAcTase antibodies and
acetylcholinesterase
(AcChoEase) antibodies, AcChoEase enzyme activity, or alpha-bungarotoxin binding indicated that ChoAcTase, AcChoEase, and AcCho receptors were colocalized at the same end plates.
...
PMID:Synthesizing enzymes for four neuroactive substances in motor neurons and neuromuscular junctions: light and electron microscopic immunocytochemistry. 612 35
A method, which is based on the use of flat, whole-mount nerve preparations, has been developed for studying the process of regenerating axon outgrowth, employing the rat sciatic nerve as a model. At various intervals after a nerve crush, animals are perfused with
aldehyde
fixatives, the nerve dissected out, and its epineurium removed. Next the nerve is flattened between two glass slides, removed and reacted (floating), then whole-mounted on a micro slide and cover-slipped. Regenerating axons have been labeled by means of the horseradish peroxidase tracing technique, a histochemical technique for
acetylcholinesterase
, or an indirect immunocytochemical technique utilizing antibodies against tubulin. With all these techniques, individual outgrowing axons and their bundles can be clearly visualized. Regenerating axons labeled by horseradish peroxidase are readily traced along their entire undulating courses from the distal margin of the crush zone to axonal tips, which mark the leading edge of several waves of outgrowing axons. It appears that such flat, whole-amount nerve preparations can be useful for obtaining: (1) accurate estimates of the rate of regenerating axon elongation, (2) values characterizing the duration of the initial delay of axonal outgrowth, and (3) information concerning the nature of axonal subpopulations that elongate at different rates.
...
PMID:Flat, whole-mount nerve preparations: a useful tool for studying the process of regenerating axon outgrowth. 636 26
The sulfide-silver method of Timm has been a widely used histochemical technique to demonstrate the presence of heavy metals in biological tissue, particularly in the central nervous system. However, the use of this method or its several modifications results in less than optimal morphological preservation and requires embedding the tissue in paraffin or freezing it and cutting it directly onto slides with a cryostat. These procedures can decrease the sensitivity and limit the application of other histochemical procedures, particularly when experiments necessitate processing large specimens or reaction procedures require techniques using free-floating sections. A perfusion-fixation protocol is described that yields sufficient fixation to cut whole frozen blocks of tissue with a sliding microtome, permits the use of free-floating sections, and allows the concurrent demonstration of horseradish peroxidase and
acetylcholinesterase
histochemistry without loss of sensitivity. The method consists of a short initial exposure to a sodium sulfide solution followed by a prolonged exposure to a combined sulfide-
aldehyde
fixative solution.
...
PMID:A perfusion-fixation procedure for the concurrent demonstration of Timm's, horseradish peroxidase (HRP), and acethycholinesterase (AChE) histochemistry. 648 Nov 50
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