PubMed Health⌕ Search

Biomedical subjects

M Brzin

Publications and source records attributed to M Brzin.

At least 37 records · Page 2Linked to original sources

Recovery of acetylcholinesterase in the diaphragm, brain, and plasma of the rat after irreversible inhibition by soman: a study of cytochemical localization and molecular forms of the enzyme in the motor end plate.

Recovery of AChE activity in the motor end plate region and end plate free region of the rat diaphragm was studied after irreversible inhibition by soman. Recovery was slow during the first 2 days and only 4 S and 10 S molecular forms of AChE were present in the end plate region. However, cytochemical evidence indicates that synaptic AChE has already started to accumulate and that the synthesis of AChE in muscle and Schwann cell might even be enhanced. Tubular structures, observed underneath the motor end plate, may serve to transport the enzyme from its sites of synthesis in the sarcoplasmic reticulum. Asymmetric molecular forms of AChE in he end plate region appeared later during recovery and, one week after poisoning, their activity was only about 50% of normal value. The limited ability of newly synthesized AChE to attach to the subcellular structures and, therefore, be retained in the muscle, may explain the phase of slow recovery. In accordance with this view, AChE activity in brain recovered in a similar way as in muscle, whereas soluble plasma cholinesterases recovered faster, apparently without a slow initial phase.

Acetylcholinesterase↗

16 S acetylcholinesterase in endplate-free regions of developing rat diaghragm.

Velocity sedimentation patterns of acetylcholinesterase (AChE, EC 3.7.1.1) in endplate-free regions of the diaphragm were studied in rats during early postnatal development. A significant amount of 16 S AChE, comprising 20% total activity, was found in endplate-free regions of the diaphragm of 8- and 19-day-old rats. By 32 days after birth, 16 S AChE accounted for less than 5% total AChE activity in endplate-free regions. 16 S AChE is, therefore, not strictly an endplate-specific molecular form. Instead, it becomes restricted to the motor endplate region of the rat diaphragm by the end of the first month of life.

Acetylcholinesterase↗

Attachment of acetylcholinesterase to structures of the motor endplate.

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 AChE in treated neuromuscular junctions. Hyaluronidase did not solubilize more AChE from isolated motor endplate regions than Ringer's solution itself. Residual AChE activity could be demonstrated histochemically in motor endplates even after the plateau of solubilization by collagenase or papain was reached. Less than 35% of junctional AChE is left after collagenase, and less than 20% after papain treatment, as estimated by the percentage of AChE activity left in the isolated endplate region of the diaphragm after protease treatment. Cytochemically, both proteases had a similar effect on postsynaptic AChE. Residual AChE activity was distributed randomly, adhering to the sarcolemma of junctional clefts. Presynaptic AChE localized in the gap between axon terminal and Schwann cell appears to be resistant to collagenase but not to papain treatment. The mode of AChE attachment or the composition of the intercellular material in this gap may differ from that of the primary and secondary clefts.

Acetylcholinesterase↗

Histochemical and cytochemical observations on the acetylcholinesterase (AChE) localization in the superior cervical ganglion (SCG) after preganglionic sympathectomy of the neonatal rat.

Results of histochemical and cytochemical reactions show that after the section of preganglionic nerve trunk performed in newborn animals, the appearance and the individual increase in AChE activity in non-innervated nerve cells follow the pattern of normally innervated ganglion cell.

Acetylcholinesterase↗

Cholinesterases and choline acetyltransferase in the longitudinal muscle of the guinea pig ileum.

In order to gain insight into the possible role of the ACh-system in the smooth muscle cell, the presence of choline acetyltransferase, acetylcholinesterase and butyrylcholinesterase was studied in the longitudinal muscle of the guinea-pig ileum after the mechanical removal of Auerbach's plexus. Such treatment completely removes all nerve elements as confirmed by histochemistry and electron-microscopic examination. It was found that in the longitudinal muscle devoid of all nervous elements a substantial percentage of the activity of all three enzymes still remained. Ultrastructural localization of acetylcholinesterase and butyrylcholinesterase was observed on the sarcolemma, sarcoplastic reticulum, nuclear membrane and invaginations of the sarcolemma. The localization of cholinesterases coincides with sites which are presumably involved in calcium movements during contraction and relaxation. It is well known that the depolarized smooth muscle responds to exogenous ACh with a reversible, calcium dependent contraction and it was suggested that ACh may act by increasing the influx of calcium through the cell membrane or by liberating calcium from its bound form. The presence of choline acetyltransferase and cholinesterase activities in the muscle cell proper, as well as the localization of cholinesterases on structures connected with calcium movements, support the coexistence of an intrinsic cholinergic mechanism in the smooth muscle.

Animals↗

Histochemical and cytochemical localization of acetylcholinesterase in retina and optic tectum of teleost fish.

The activity of cholinesterase and its cellular and subcellular localization were investigated in the retina and optic tectum of Eugerres plumieri and in the retina of Carassius carassius by means of radiometric, histochemical, and cytochemical procedures. In both fishes only the presence of acetylcholinesterase could be demonstrated. This study, besides confirming previous findings that acetylcholinesterase is located in the ganglion and amacrine cells of the retina as well as in the inner plexiform layer, in addition provides evidence that the enzyme is also present at the region of photoreceptor synapses between the cell bodies and apposing extensions of the horizontal cells of the same layer. The latter localization may indicate the involvement of a cholinergic mechanism at the functional contacts (transferapses) between the horizontal cells. In the optic tectum of Eugerres plumieri, histochemistry reveals fine distinguishable bands of acetylcholinesterase activity; two of the bands are quite sharply defined, whereas three others have rather a more diffuse appearance. The presence of these bands and their distribution may suggest a widespread distribution of cholinergic elements in the optic tectum.

Acetylcholinesterase↗

Quantitative evaluation of the trapping reaction of copperthiocholine histochemical procedures for localization of cholinesterases.

The aim of the present study was to investigate whether the trapping reaction of the histochemical procedure for the localization of ChE of Koelle and Friedenwald (1949) and its modification by Brzin and Pucihar (1976) proceeds quantitatively. The weight of the precipitate formed in the tissue sample during the histochemical procedure was compared with enzyme activity of an equal sample. The differential magnetic microbalance was used for measurements of reduced weight and for previous determination of density of the precipitate. The evidence for the composition of the final product was drawn from the quantitative analysis of copper and iodine and from the infrared spectra. Tsuji's statement (1974) that cuprous copper thiocholine iodide is the final product of histochemical procedures investigated was confirmed. It was found that the trapping reaction of the original as well as of the modified procedure under our experimental conditions in tissue sections proceeds quantitatively which means that one of the basic conditions for reliable localization is fulfilled.

Animals↗

A comparison between the one-step and the two-step copper thiocholine procedure for the cytochemical localization of cholinesterases.

A comparison between the one-step and the two step copper thiocholine procedure for the subcellular localization of cholinesterases was undertaken. The results indicate that only under experimental conditions with short incubation times and precise control of the conversion into sulphide is the localization of the primary precipitate and that of the secondary precipitate identical. It was concluded that the conversion of the primary precipitate into Cu-sulphide is not necessary and can lead to artefacts.

Animals↗

Cytochemical localization of acetylcholinesterase in the rat striatum.

An attempt was made to determine the subcellular localization of AChE in rat striatum with several Cu-thiocholine procedures using slices of intact tissue and in isolated organelles obtained by gradient centrifugation. Regardless the method used the reaction product, indicating the localization of AChE activity shows the usual intracellular and extracellular distribution. Although AChE activity was present at the extrajunctional axolemma of the nerve endings almost no enzyme activity was observed in the junctional region either in tissue sections or on isolated synaptosomes.

Acetylcholinesterase↗

Iodide, thiocyanate and cyanide ions as capturing reagents in one-step copper-thiocholine method for cytochemical localization of cholinesterase activity.

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.

Animals↗