Biomedical subjects

J J Plomp

Publications and source records attributed to J J Plomp.

24 records · Page 2Linked to original sources

PubMed1995

Acetylcholine release in myasthenia gravis: regulation at single end-plate level.

In myasthenia gravis, loss of acetylcholine receptors at motor end-plates is induced by antireceptor autoantibodies. At end-plates in rats in which myasthenia gravis-like symptoms are induced by chronic treatment with alpha-bungarotoxin, acetylcholine release is increased. Within muscles from such rats there is a strong correlation between the increase of acetylcholine release at an end-plate and the loss of postsynaptic acetylcholine receptors, caused by the toxin. The question is whether upregulation of acetylcholine release is a clinically relevant compensatory mechanism in myasthenia gravis or only a feature of the animal model using alpha-bungarotoxin. We investigated electrophysiologically the in vitro acetylcholine release at end-plates of muscles from patients with myasthenia gravis and rats with experimental autoimmune myasthenia gravis where acetylcholine receptor reduction is caused by autoantibody attack. In both human and rat autoimmune myasthenic muscle, the mean quantal content was considerably increased compared with control levels. At each individual myasthenic end-plate, the increase in quantal content appeared to be correlated with the reduction of the amplitude of the miniature end-plate potential. This finding suggests the existence of an important compensatory mechanism in myasthenia gravis, in which retrograde acting factors (i.e., from muscle fiber to nerve terminal) upregulate acetylcholine release.

Acetylcholine↗

PubMed1995

Acquired neuromyotonia: evidence for autoantibodies directed against K+ channels of peripheral nerves.

Acquired neuromyotonia is characterized by hyperexcitability of motor nerves leading to muscle twitching, cramps, and weakness. The symptoms may improve following plasma exchange, and injection of immunoglobulin G (IgG) from 1 neuromyotonia patient into mice increased the resistance of neuromuscular transmission to d-tubocurarine. Here we examine nerves and muscle in vitro from mice injected with plasma or purified IgG from 6 neuromyotonia patients or pooled control subjects, and cultured dorsal root ganglion cells after treatment with IgG. Three of the patients had antibodies against human voltage-gated potassium channels labeled with 125I-alpha-dendrotoxin. The quantal release of acetylcholine (quantal content) at end-plates in diaphragms from mice treated with neuromyotonia IgG preparations was increased by 21% relative to control values (p = 0.0053). With one IgG preparation, the duration of the superficial peroneal nerve compound action currents was increased by 93%. The dorsal root ganglion cells treated with this IgG showed a marked increase in repetitive firing of action potentials. All effects were similar to those obtained with aminopyridines. We conclude that at least some patients with acquired neuromyotonia have antibodies directed against aminopyridine- or alpha-dendrotoxin-sensitive K+ channels in motor and sensory neurons, and they are likely to be implicated in the disease process.

4-Aminopyridine↗

PubMed1994

The upregulation of acetylcholine release at endplates of alpha-bungarotoxin-treated rats: its dependency on calcium.

1. The presynaptic component of an adaptive feedback mechanism leading to increased acetylcholine (ACh) release was studied in endplates of diaphragms from rats treated chronically with alpha-bungarotoxin (alpha BTX). 2. Quantal contents were calculated 'directly' from the amplitude of miniature endplate potentials (MEPPs) and endplate potentials (EPPs) which were recorded after mu-conotoxin treatment to prevent muscle action potentials. 3. In vitro application of the Ca2+ channel blockers nifedipine (10 microM) or omega-conotoxin (40 nM) had no significant effect on the increased quantal content of endplates from alpha BTX-treated rats. 4. At control endplates, in vitro block of presynaptic K+ channels by 5 microM 3,4-diaminopyridine did increase the quantal content to a level which was similar to that found in endplates of alpha BTX-treated rats but also induced a broadening of EPPs, which was not found at endplates after alpha BTX treatment. 5. The difference between quantal contents of alpha BTX-treated and control rats was highly dependent on the [Ca2+]o/[Mg2+]o ratio when [Mg2+]o was fixed at 1 mM. At low [Ca2+]o, the quantal content of endplates from alpha BTX-treated rats was lower than that of controls while at [Ca2+]o in the normal and high range this was reversed. However, changing the [Ca2+]o/[Mg2+]o ratio by means of [Mg2+]o, at a fixed [Ca2+]o of 2 mM, did not influence the relative increase of quantal contents at endplates from alpha BTX-treated rats. Double logarithmic plots of the 'toxin-induced' myasthenia gravis (TIMG) and control quantal content versus [Ca2+]o had an approximately linear part between 0.2 and 1.5 mM [Ca2+]o. The slopes of the TIMG and control lines were 1.81 and 0.96, indicating that the ACh release in TIMG muscles was more sensitive to changes of [Ca2+]o than controls. 6. At normal [Ca2+]o and [Mg2+]o, the depression of EPP amplitude during stimulation of the phrenic nerve at 30-50 Hz was somewhat larger at endplates from alpha BTX-treated rats than at control endplates. At low [Ca2+]o, the potentiation of EPP amplitudes during a stimulus train was much larger at endplates from alpha BTX-treated rats than from controls. 7. The results do not support the idea that the increased release of ACh is caused via regulatory effects on the presynaptic Ca2+ or K+ channels. Instead, the anomalous dependency of ACh release on Ca2+ in muscles of alpha BTX-treated rats suggests that a cytoplasmic, Ca(2+)-dependent, component is involved in the adaptive change of transmitter release.

Acetylcholine↗

PubMed1992

Adaptation of quantal content to decreased postsynaptic sensitivity at single endplates in alpha-bungarotoxin-treated rats.

1. Rats were injected once every 48 h with alpha-bungarotoxin (alpha BTX) for periods up to 6 weeks. Injections caused weakness of facial muscles which lasted about 8 h. Hemidiaphragms were dissected for biochemical and electrophysiological measurements. 2. In muscles from animals treated for 2-3 weeks with toxin, the binding of 125I-alpha BTX was reduced to 58%, and the ACh content to 81% of control values. Choline acetyltransferase activity was unchanged. ACh release evoked by 3 Hz nerve stimulation was increased to 175% of control values. 3. The use of mu-conotoxin, which specifically blocks muscle action potentials, enabled the recording of full-sized endplate potentials (EPPs) and miniature endplate potentials (MEPPs) at normal muscle membrane potentials (-70 to -80 mV). The amplitude of MEPPs was decreased to 57% in muscles from animals treated for 3 weeks with alpha BTX. The mean of the quantal contents, calculated from the ratio of the corrected EPPs and the MEPPs, was increased to 154%. 4. Within individual muscles of both alpha BTX-treated and control rats, there was an inverse relationship between the quantal content of an endplate and its MEPP amplitude. 5. The MEPP frequency of endplates from control muscles was positively correlated with the quantal content. However, this correlation was not found in alpha BTX-affected muscles. 6. Three hours after a single injection of alpha BTX the amplitude of the MEPPs was reduced to about 60% of control values but no increase of the quantal content was found. During the first few days of alpha BTX treatment the quantal content gradually increased; it reached a plateau between 20 and 30 days. 7. The results suggest the existence of an adaptive mechanism, operating at individual endplates, in which retrograde signals at the motor nerve terminals modulate ACh release when neuromuscular transmission is endangered by block of acetylcholine receptors.

Acetylcholine↗

PubMed1991

A non-immunogenic myasthenia gravis model and its application in a study of transsynaptic regulation at the neuromuscular junction.

A non-immunological model for myasthenia gravis was developed in rats: 'toxin-induced myasthenia gravis'. Rats were injected once every 48 h with 3-5 micrograms alpha-bungarotoxin for periods of up to 5 weeks. This treatment caused weakness, especially of facial muscles. Respiration, however, was unaffected. Miniature endplate potentials and 125I-alpha-bungarotoxin binding in the extensor digitorum longus muscles were severely reduced. Acetylcholine release evoked by electrical and chemical (50 mM KCl) stimulation was higher in diaphragms from alpha-bungarotoxin-treated rats than in those from control animals. Histological investigation of the tibialis anterior muscle provided no evidence that the endplates were enlarged. It is concluded that the activity of acetylcholine receptors influences the rate of transmitter release in the neuromuscular junction and it is suggested that a transsynaptic regulation process may be active in myasthenia gravis. The present animal model for myasthenia gravis seems very suitable for studying such a regulation of transmitter release.

Acetylcholine↗

PubMed1989

Kinetics of the membrane current mediated by serotonin 5-HT3 receptors in cultured mouse neuroblastoma cells.

1. Ionic currents mediated by serotonin 5-HT3 receptors were studied in the mouse neuroblastoma cell line N1E-115, using suction pipettes for intracellular perfusion and voltage clamp recording. The dependence of the kinetics of the membrane current on serotonin concentration was investigated. 2. At a holding potential of -70 mV application of 5-HT (5-hydroxytryptamine creatinine sulphate) causes a transient inward current. The i-V curve of the peak amplitude is linear between -80 and 60 mV. The reversal potential is 20 +/- 4 mV (mean +/- S.D.). The kinetics of the transient ionic current are independent of the holding potential. 3. In the presence of 5-HT the membrane current decays to a small steady-state level with a single-exponential time course. The time constant of decay decreases with increasing concentration of the agonist, to a minimum value of 6.5 +/- 1.5 s for concentrations of 5-HT greater than or equal to 3 microM. 4. When the agonist is rapidly removed, single-exponential decay of the ionic current is observed. The time constant of this decay in the absence of 5-HT amounts to 6.9 +/- 1.5 s and is independent of the membrane potential and of the concentration of 5-HT used. 5. In the presence of low concentrations of 5-HT the peak amplitude of the inward current evoked with a high concentration of agonist is gradually reduced. The onset of this desensitization follows the same time course as the decay of the membrane current. In the range from 0.7 to 1.5 microM-5-HT both kinetic processes show the same steep concentration dependence. 6. Recovery from desensitization, measured at variable intervals after removal of the agonist, can be fitted by a single-exponential function with a time constant of 18 +/- 4 s. 7. The results show that the kinetic properties of the 5-HT3 receptor-mediated ionic current can only be described by a complex, co-operative model.

Action Potentials↗