PubMed Health⌕ Search

Biomedical subjects

U Musshoff

Publications and source records attributed to U Musshoff.

At least 37 records · Page 2Linked to original sources

Membrane currents elicited by the organic calcium channel blocker verapamil in native and rat brain RNA-injected oocytes of Xenopus laevis.

For further analysis of the action of the diphenylalkylamine verapamil (CAS 152-11-4), the ability of verapamil to elicit membrane currents by itself was investigated in native and rat brain. RNA-injected oocytes of Xenopus laevis. Administration of verapamil elicited inward currents which remained constant or increased slightly during ongoing application. In native and RNA-injected oocytes the current responses were similar in shape, but larger in size in RNA-injected oocytes. The currents increased up to the maximal tested concentration of 1 mmol/l verapamil; the threshold concentration was below 80 mumol/l. After removal of follicular tissues the verapamil response was nearly doubled. During verapamil administration the input resistance was increased up to 1.7 of the initial value. The current response to verapamil can be subdivided into an early and late component. The equilibrium potential of the early component ranged between -80 and -110 mV; the late component which increased slightly during verapamil application, had an equilibrium potential between 0 and -20 mV. Under the influence of potassium channel blockers (tetraethylammonium and cesium chloride) or chloride channel blockers (anthracene-9-carbonic acid and the indanyloxy-acetic acid derivative IAA-94) the verapamil induced currents were reduced. Thus, the results indicate that beside the calcium channel-blocking effect, verapamil can induce currents by itself, presumably by acting on the potassium and chloride leakage.

Animals↗

Mechanism of action of the epileptogenic drug pentylenetetrazol on a cloned neuronal potassium channel.

The action of the epileptogenic agent pentylenetetrazol (PTZ) on a cloned potassium channel of the rat brain was studied. The Kv1.1 channel was expressed in oocytes of Xenopus laevis and potassium currents were investigated in outside-out and inside-out membrane patches. The results show that PTZ increased the multi-channel potassium currents at strongly negative potentials and decreased them at potentials positive to -35 mV both in outside-out and inside-out membrane patches. The extent and manner of PTZ action, the concentration dependence as well as the onset and time course of the PTZ effect were the same both in outside-out and inside-out membrane patches. The single-channel potassium currents showed an increase in open probability and frequency of opening and a decrease in close time at -50 mV and vice versa at 0 mV with application of PTZ. The amplitude of single-channel current, the open time and the latency to the first channel opening remained almost unchanged under PTZ. The results indicate that PTZ acts via the cell membrane and influences the membrane-associated part of the potassium channel. Thereby, PTZ accelerates the transition from the inactivated to the open state of the channel at strongly negative potentials and reduces it at slightly negative and positive potentials. This mechanism may be the basis for a gate function which is in favour of the development of epileptic discharges.

Animals↗

Expression and functional characterization of a melatonin-sensitive receptor in Xenopus oocytes.

Melatonin (MEL) plays a central role in the regulation of seasonal cycles and in the control of circadian rhythms in mammals. Functional MEL-sensitive receptors were expressed in Xenopus laevis oocytes following injection of poly (A)+ RNA from rat brain. Administration of 0.1-100 micromol/l MEL to voltage-clamped oocytes (holding potential: -70 mV) elicited oscillatory inward currents (reversal potential: -24 mV) which could be blocked by 9-anthracenecarboxylic acid and caffeine. After preincubation with pertussis toxin (PTX) the MEL response disappeared. The expressed MEL-sensitive receptor probably activates Ca(2+)-dependent chloride currents via a PTX-sensitive G protein and the phosphoinositol pathway.

Animals↗

Effects of pentylenetetrazol on GABA receptors expressed in oocytes of Xenopus laevis: extra- and intracellular sites of action.

The convulsive agent pentylenetetrazol (PTZ) antagonized gamma-aminobutyric acid (GABA)-induced membrane currents on RNA-injected Xenopus oocytes with both extra- and intracellular applications. With extracellular administration PTZ enters the cell within a few minutes and reaches concentrations in the millimolar range. The permeability of the plasma membrane makes it possible for systemically applied PTZ to elicit its effect on the GABA-induced currents via extra- as well as intracellular sites of action.

Animals↗

Activation of ATP-sensitive potassium channels in follicle-enclosed xenopus oocytes by the epileptogenic agent pentylenetetrazol.

For further investigation of the epileptogenic properties of pentylenetetrazol (PTZ), membrane currents elicited by PTZ were analysed in native Xenopus oocytes. PTZ elicited a sequence of membrane currents. Two inward currents have been described to be due to a decrease in potassium permeability and an increase in chloride permeability. Experiments performed up to 3 days after preparation of the oocytes showed that PTZ is also able to activate an outward current. This current is: (1) reversed near the potassium equilibrium potential, (2) associated with a decrease in membrane resistance, (3) reduced by tetraethylammonium and caesium, (4) abolished by defolliculation, and (5) blocked by glibenclamide. Thus, the current can be interpreted to be due to an activation of ATP-sensitive potassium (KATP) channels located in the follicle cells. An activation of KATP channels by PTZ may contribute to termination and re-initiation of seizure activity.

Adenosine Triphosphate↗

Prediction of neurotoxic potency of hazardous substances with a modular in vitro test battery.

Neurotoxic action was investigated on different model nervous systems linked to a modular in vitro test battery. Voltage operated potassium channels and glutamate operated ion channels expressed in oocytes of the clawed frog Xenopus laevis by injection of cRNA (cloned RNA) or mRNA, respectively, as well as isolated neurons and isolated neuronal networks from the buccal ganglia of the snail Helix pomatia, were used as consecutive modules of different complexity. Lead (Pb2+) was chosen as a known neurotoxic model substance to evaluate the suitability of the test battery to predict the neurotoxic potency of hazardous substances, to establish dose-response relationships, and to investigate the basic mechanisms involved in neurotoxicity. All modules delivered consistent results: potassium currents were reduced by lead with a threshold concentration of 0.1 mumol/l. Membrane currents elicited by the glutamate receptor agonists kainate were decreased by lead with a threshold concentration below 0.1 mumol/l, while currents elicited by the agonist AMPA were not affected. Action potentials generated by the isolated B4 snail neuron showed a decrease of potential amplitude and a prolongation of potential duration after application of lead. The neuronal network controlling the feeding activities of the snail reacted with a decrease of the frequency of the spontaneously generated feeding depolarisations, thus showing the direct neurotoxic effect of lead on body functions and behaviour.

Animals↗

Lead-induced blockage of kainate-sensitive receptor channels.

The effects of bivalent lead on ion channels activated by kainate and alpha-amino-3-hydroxy-5-methyl-4-isoxazolpropionate (AMPA) were studied using Xenopus oocytes microinjected with mRNA from rat brain. Lead reduced kainate-induced membrane currents in a reversible and dose-dependent manner, without affecting membrane currents induced by AMPA. Lead decreased the kainate currents with a concentration of 0.1 micromol/1 to 0.93 +/- 0.01 and with a concentration of 100 micromol/1 to 0.41 +/- 0.04 of the control values. The blocking effect of lead on kainate responses was voltage dependent. The inhibition was strongest at -90 mV to -70 mV and became weaker at more positive membrane potentials. The effect of lead on the kainate-induced membrane currents remained unchanged when the concentration of kainate was increased. Hence lead probably represents a noncompetitive channel-blocking agent for non-N-methyl-D-aspartate (NMDA) receptor channels activated by kainate.

Animals↗

Effects of lead on cloned voltage-operated neuronal potassium channels.

The action of lead (Pb2+) on cloned voltage-operated potassium channels of the rat brain was investigated in oocytes of Xenopus laevis. Pb2+ was found to decrease the potassium currents. This effect was due to a shift of the current-voltage relation in a positive direction (up to 30 mV). The Pb2+ effect appeared at a threshold concentration of about 0.1 mumol/l and was maximal at a concentration of about 30 mumol/l. At a potential of -30 mV, the concentration needed for a 50% reduction of the potassium current was 1.0 mumol/l. The depressant effect of Pb2+ was obtained with all potassium channels tested (Kv1.1, Kv1.2, Kv1.4, Kv2.1, Kv3.4). It was minimal for the Kv2.1 channel and maximal for the Kv1.1 channel at potentials negative to 0 mV. An effect comparable with that of Pb2+ could not be induced by the application of magnesium or calcium. The external application of Pb2+ led to a decrease of potassium currents in outside-out but not in inside-out membrane patches. Overall, Pb2+ had a significant effect on the potassium channels which may contribute to the mechanisms of Pb2+ neurotoxicity.

Animals↗

Improvement and testing of a concentration-clamp system for oocytes of Xenopus laevis.

A system for rapid solution exchange on Xenopus laevis oocytes in the two-electrode voltage-clamp mode (Madeja et al., 1991) was improved for investigations on oocytes with removed follicular tissues. The speed of application and the time for potassium ions to reach the oocyte membrane (t50) was found to be about 40 ms in oocytes without follicular tissues and 360 ms for oocytes with follicular tissues. This rate of solution exchange is fast enough to measure the fast desensitizing component of the AMPA current response.

Animals↗

Effects of the epileptogenic agent strychnine on membrane currents elicited by agonists of the NMDA and non-NMDA receptors in Xenopus oocytes.

The effects of strychnine (STRY) on ion channels activated by N-methyl-D-aspartate (NMDA), kainate (KA), alpha-amino-3-hydroxy-5-methyl-4-isoxazolpropionate (AMPA) and quisqualate (QUIS) were studied using Xenopus oocytes, microinjected with mRNA from rats' brains. STRY reduced NMDA-, KA- and AMPA-induced membrane currents in a dose-dependent manner. The effect was more pronounced with NMDA than with KA and AMPA. QUIS-induced membrane currents were not affected by STRY. The depressive effect of STRY on NMDA responses was voltage dependent. The effect of STRY on the NMDA-induced membrane currents remains unchanged when the concentration of NMDA or glycine was increased. Intracellular injection of STRY did not alter the NMDA response.

Animals↗

Decrease and increase of responses to glutamate receptor agonists in RNA-injected Xenopus oocytes by the epileptogenic agent pentylenetetrazol: dependence on the agonist concentration.

The effects of the epileptogenic agent pentylenetetrazol (PTZ) on current responses to glutamate (Glu) and to the Glu receptor agonist N-methyl-D-aspartate (NMDA), kainate (KA), alpha-amino-3-hydroxy-5-methyl-4-isoxazole propionic acid (AMPA) and quisqualate (QA) were studied on RNA-injected Xenopus oocytes. PTZ antagonized the reactions to Glu as well as to NMDA, KA, AMPA and QA. With increasing concentration of AMPA, however, the depressive effect of PTZ turned to an augmentation. The complete change from a decreasing to an increasing effect of PTZ with elevated agonist concentration was unique for current responses by AMPA.

Animals↗

Potassium currents in epilepsy: effects of the epileptogenic agent pentylenetetrazol on a cloned potassium channel.

The effect of the epileptogenic agent pentylenetetrazol (PTZ) on the cloned rat brain potassium channel Kv1.1 (labelled also RCK1) was investigated in the Xenopus laevis oocyte expression system. The Kv1.1 channel was affected by PTZ in a voltage-dependent manner. PTZ increased the potassium currents at more negative potentials and decreased them at more positive potentials. At a potential of -50 mV the potassium currents were increased by 0.97 and at -20 mV decreased by 0.21 of control value with 100 mmol/l PTZ. The potential at which the inversion from increase to decrease occurred was -33 mV. The inactivation characteristic of the current was shifted to more negative potentials by PTZ. The PTZ effect was obtained at a threshold concentration of 1 mmol/l and increased with rising PTZ concentrations. After removal of the tissues covering the oocyte membrane, the PTZ effect was augmented; with a concentration of 10 mmol/l PTZ the potassium currents at 0 mV were decreased by 0.04 in oocytes with covering tissues and by 0.27 of control value in oocytes without covering tissues. Under current-clamp conditions, PTZ decreased small depolarizations and increased larger depolarizations. This effect of PTZ represents a 'discriminatory function' that may contribute to epileptogenesis in nervous tissues.

Animals↗

Effects of the epileptogenic agent bicuculline methiodide on membrane currents induced by N-methyl-D-aspartate and kainate (oocyte; Xenopus laevis).

The actions of bicuculline methiodide (BIC) on N-methyl-D-aspartate (NMDA)- and kainate (KA)-activated ion channels were studied using Xenopus oocytes, previously microinjected with RNA from rat brains. BIC reduced NMDA- and, with a lower efficiency, KA-induced membrane currents in a dose-dependent manner. The BIC effect on both agonist responses showed a voltage dependency with a lower grade of reduction at more positive holding potentials. By increasing the concentrations of the agonists, the reduction of the agonist-induced membrane currents by BIC was increased.

Animals↗

Suppression of a ligand operated membrane current by the epileptogenic agent pentylenetetrazol in oocytes of Xenopus laevis after injection of rat brain RNA.

The effects of the epileptogenic substance pentylenetetrazol (PTZ) on ligand operated membrane channels were studied. For this purpose serotonin (5-HT) sensitive channels were expressed in RNA injected oocytes of Xenopus laevis. With simultaneous application of both substances, the response to 5-HT was reduced and eventually abolished by PTZ with increasing concentrations (5-100 mM). A reduction of the 5-HT response also appeared when PTZ was applied in various intervals (15 and 240 s) before 5-HT. It may be assumed that PTZ produces a component of its epileptogenic effect by acting on ligand operated membrane channels.

Animals↗

Tunicamycin-induced inhibition of functional expression of glutamate receptors in Xenopus oocytes.

The effects of tunicamycin, a specific inhibitor of N-linked glycosylation, on functional expression of glutamate receptor subtypes were investigated in RNA-injected oocytes. In the presence of tunicamycin the expression of ligand-operated receptors sensitive to kainate, alpha-amino-3-hydroxy-5-methyl-4-isoxazolepropionate (AMPA) and quisqualate were completely blocked. The inhibitory effect was reversible after removal of tunicamycin from the culture medium.

Animals↗

Time course of glutamate receptor expression in individual oocytes of Xenopus laevis after injection of rat brain RNA.

1. The receptor for the neurotransmitter glutamate was functionally expressed in oocytes of Xenopus laevis after microinjection of rat brain RNA. The functional differentiation of this receptor type was further analyzed. 2. The development of the sensitivity to the agonists showed a time course which was differential for the various ligands in individual oocytes. 3. Sensitivity appeared after one day for kainate (KA), after two days for alpha-amino-3-hydroxy-5-methyl-4-isoxazole-propionate (AMPA) and quisqualate (QA), and after five days for N-methyl-D-aspartate (NMDA). 4. The KA response was markedly reduced by simultaneous application of AMPA. This was even found on the first day when an agonistic AMPA reaction was not detectable. 5. NMDA and non-NMDA receptors can clearly be differentiated by their delay of expression.

Animals↗

Membrane currents elicited by the epileptogenic drug pentylenetetrazole in the native oocyte of Xenopus laevis.

The effects of the epileptogenic agent pentylenetetrazole (PTZ) on membrane currents of native oocytes of Xenopus laevis were studied. PTZ elicits a response that consists of two inward currents. The first one is interpreted to be due to a decrease of potassium permeability since: (1) the input resistance is increased; (2) the equilibrium potential is near that of potassium; (3) the current is decreased during administration of potassium channel blocking agents; and (4) the PTZ response can be mimicked by blocking potassium channels without PTZ application. The second one is interpreted to be due to an increase of chloride permeability since: (1) the input resistance is decreased; (2) the equilibrium potential is near that of chloride; and (3) the response is decreased during administration of chloride blocking agents. These findings correspond to some extent with those made in neurons.

Animals↗

Inward currents elicited by stream of fluid during transmitter-induced current oscillations in RNA-injected oocytes of Xenopus laevis.

A stream of fluid elicited an inward current in RNA-injected oocytes of Xenopus laevis during transmitter-induced current oscillations (stream evoked inward current, Ii,st). The Ii,st showed the following characteristics: (1) amplitude and duration (half width time) ranged between 10 and 300 nA and 1 and 3 s, respectively. (2) The Ii,st could be evoked only during transmitter-induced current oscillations; with blockade of the oscillations the Ii,st disappeared. (3) The induction of the Ii,st was independent of the composition of the washing fluid.

Animals↗