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M Thieffry

Publications and source records attributed to M Thieffry.

At least 37 records · Page 2Linked to original sources

The effect of calcium ions on the glutamate response and its desensitization in crayfish muscle fibres.

The responses of crayfish muscle fibres to bath application or long ionophoresis of L-glutamate were studied in normal and low Ca2+ solutions. The smaller responses recorded in low Ca2+ solutions have characteristics suggesting a faster desensitization. Desensitization and recovery have complex kinetics. Desensitization is faster and recovery slower when external Ca2+ concentration is reduced. Both components of the recovery phase, which can be fitted by the sum of two exponentials, are affected by the external Ca2+ concentration. Recovery can be accelerated by external Ca2+ ionophoresis onto desensitized glutamate receptors. Responses to brief glutamate pulses of low intensity are not affected by Ca2+ reduction. For higher intensities, signs of desensitization are detectable early in the rising phase of the response. Concanavalin A (Con A) blocks both desensitization and Ca2+ dependence with similar time courses. Whether or not the preparation has been treated with Con A, the slowly rising responses recorded in isotonic Ca2+ do not show signs of desensitization. Con A causes a partial blockade of the glutamate response. The Ca2+ dependence of the glutamate response can be explained by the Ca2+ dependence of the desensitization process, the cation acting at ectocellular sites of the muscle membrane.

Animals↗

Binding of a Glycera convoluta neurotoxin to cholinergic nerve terminals triggers a Ca-dependent acetylcholine release.

The venom glands of the annelid Glycera convoluta contain a neurotoxin which triggers ACh release from frog motor terminals and Torpedo synaptosomes. This neurotoxin binds to presynaptic, but not postsynaptic plasma membranes prepared from Torpedo electric organ. The binding site is an ectocellularly oriented protein. The binding does not require Ca. It is inhibited by pretreatment of the membrane by Concanavalin A. The toxin induced ACh release is Ca-dependent and inhibited by D 600.

Acetylcholine↗

Binding of a Glycera convoluta neurotoxin to cholinergic nerve terminal plasma membranes.

The crude extract of venom glands of the polychaete annelid Glycera convoluta triggers a large Ca2+-dependent acetylcholine release from both frog motor nerve terminals and Torpedo electric organ synaptosomes. This extract was partially purified by Concanavalin A affinity chromatography. The biological activity was correlated in both preparations to a 300,000-dalton band, as shown by gel electrophoresis. This confirmed previous determinations obtained with chromatographic methods. This glycoprotein binds to presynaptic but not postsynaptic plasma membranes isolated from Torpedo electric organ. Pretreatment of intact synaptosomes by pronase abolished both the binding and the venom-induced acetylcholine release without impairing the high K+-induced acetylcholine release. Pretreatment of nerve terminal membranes by Concanavalin A similarly prevented the binding and the biological response. Binding to Torpedo membranes was still observed in the presence of EGTA. An antiserum directed to venom glycoproteins inhibited the neurotoxin so we could directly follow its binding to the presynaptic membrane. Glycera convoluta neurotoxin has to bind to a ectocellularly oriented protein of the presynaptic terminal to induce transmitter release.

Acetylcholine↗

Concanavalin A blocks the Ca2+ -dependence of crayfish muscle fiber responses to glutamate.

(1) The response of crayfish muscle fibers to bath-applied glutamate is strongly inhibited when the Ca concentration of the physiological solution is reduced. Other divalent cations cannot substitute for Ca. The trivalent impermeant cation La can at low concentration replace Ca. Moreover, decreasing the Ca concentration in the presence of La potentiates the glutamate response. (2) The time course of responses to ionophoretically applied glutamate suggests a faster desensitization in low Ca solutions. The lectin concanavalin A, which blocks desensitization, also eliminates the decrease of the glutamate response in low Ca solutions. (3) The above results are compared to available data concerning Ca-dependence, desensitization and effects of concanavalin A.

Animals↗

Partial purification of the Glycera convoluta venom components responsible for its presynaptic effects.

The crude extract of glands appended to the jaws of the polychaete annelid Glycera convoluta induces an important increase in the spontaneous quantal transmitter release on frog and crayfish neuromuscular junctions and on Torpedo nerve-electroplaque junctions. The venom similarly triggers acetylcholine (ACh) release from synaptosomes purified from Torpedo electric organ. At the frog neuromuscular junction, the reproducibility, the reversibility and the dose-dependence of the venom action permit a quantitative evaluation of the effect. The crude venom extract has been fractionated by gel-filtration. The effect on transmitter release has been found in a high molecular weight fraction distinct from those which contain the protease and phospholipase activities.

Acetylcholine↗

Effects of high calcium solutions on glutamate sensitivity of crayfish muscle fibres.

Crayfish neuromuscular preparations were studied after 18--36 h exposure to high calcium solutions. As previously reported for frog neuromuscular preparations the treatment damaged the nerve terminals and decreased junctional potentials. The resting potentials and input resistances of the muscle fibres were not affected; but their sensitivity to glutamate was significantly decreased when compared to that of control muscles. After exposure to high calcium, the sensitivity to gamma-aminobutyric acid, the putative transmitter at inhibitory synapses, was increased. Apparently normal twitches were elicited by direct stimulation, and calcium spikes could still be observed in the fibres. A decreased sensitivity to glutamate was also noted in experiments carried out on denervated muscles 8 months after section of the motor axons. Possible relations between nerve terminal damage and the decrease in sensitivity to glutamate are discussed.

Animals↗

Effect of the venom of Glycera convoluta on the spontaneous quantal release of transmitter.

A neurotoxin able to increase the spontaneous release of transmitter was found in the venom glands of the polychaete annelid Glycera convoluta. We studied the effect of this venom on the frog cutaneous pectoris muscle, where its application produced a prolonged (20-h), high-frequency discharge of miniature potentials. After 5 h of action, the initial store was renewed several times but no detectable ultrastructural changes were observed. After 19 h of sustained activity, nerve terminals with their normal vesicular contents were infrequent; others were fragmented and contained swollen mitochondria, abnormal inclusions, and vesicles of various sizes. In the noncholinergic crayfish neuromuscular preparation, the venom triggered an important increase in spontaneous quantal release that subsided in 1 h. An activity higher than that in resting conditions then persisted for many hours. This high electrical activity was not accompanied by any detectable structural modifications after 3 h. In the torpedo electric organ preparation, the venom elicited a burst of activity that returned to control levels in 1 h. The release of ACh (evaluated by the efflux of radioactive acetate) paralleled the high electrical activity. No morphological changes or significant depletion of tissue stores were detected. The venom of Glycera convoluta appears to enhance considerably the release of transmitter without impairing its turnover. The venom effect is Ca++ dependent and reversible by washing, at least during the first hour of action. Because the high rate of transmitter release appears dissociated from the later-occurring structural modifications, it is possible that the venom mimics one component of the double mode of action proposed for black widow spider venom.

Animals↗

The presynaptic action of L-glutamate at the crayfish neuromuscular junction. Results obtained by intra-axonal recordings and nerve terminal damage.

The presynaptic action of glutamate was investigated in crayfish neuromuscular preparations. The excitability of locally stimulated nerve terminals increased during the perfusion of glutamate. Electrotonically propagated depolarizations were recorded in the excitatory axons when glutamate was ionophoretically applied to sensitive spots on the muscle membrane. Depolarizations of the axon were also obtained when glutamate, GABA and aspartate were added to the bath. In order to evaluate the part of the presynaptic effect in the depolarization induced in muscle fibres, nerve terminals were damaged either by treatment with high-Ca solutions or by denervation. In both cases, a decreased sensitivity to glutamate was found. Possible relationships between the nerve terminal damage and the change in the response to glutamate are proposed.

Animals↗