PubMed Health⌕ Search

Biomedical subjects

W Schreibmayer

Publications and source records attributed to W Schreibmayer.

16 recordsLinked to original sources
PubMed1992

Membrane actions of calcitonin gene-related peptide in cardiac and smooth muscle myocytes.

Calcitonin gene-related peptide is a 37-amino acid neuropeptide acting as a transmitter of nonadrenergic, noncholinergic nerves in the heart. Binding sites of high affinity have been reported in coronary arteries, in atria, and, of minor density, in ventricular myocardium. These sites are likely linked to G-proteins mediating modifications of ion channel opening probability and duration and to stimulation of adenylate cyclase activity and cAMP-mediated alterations of ion channel activities. In isolated and perfused guinea pig hearts, low concentrations of CGRP (1-3 nM) exerted no chronotropic effect, but increased coronary flow slightly. Atrioventricular conduction duration and effective refractory period of atrioventricular conduction were prolonged by 3 nM of CGRP. The higher concentration of 10 nM increased the sinus rate, and the effects on the atrioventricular node were counterbalanced. HV and QRS duration of the ECG remained essentially unchanged, but persistent ventricular fibrillation was inducible by burst stimulation in all CGRP-treated hearts. Results in human myometrial myocytes indicate that CGRP exerted direct G protein-mediated activation of potassium channels, leading to hyperpolarization and smooth muscle relaxation. Activation of potassium channels, most prominent in smooth muscle relaxation, is likely an additional factor in the cardiostimulatory profile of CGRP.

Action Potentials↗
PubMed1992

Level of expression controls modes of gating of a K+ channel.

Several distinct subfamilies of K+ channel genes have been discovered by molecular cloning, however, in some cases the structural differences among them do not account for the diversity of K+ current types, ranging from transient A-type to slowly inactivating delayed rectifier-type, as members within each subfamily have been shown to code for K+ channels of different inactivation kinetics and pharmacological properties. We show that a single K+ channel cDNA of the Shaker subfamily (ShH4) can express in Xenopus oocytes not only a transient A-type K+ current but also, upon increased level of expression, slowly inactivating K+ currents with markedly reduced sensitivity to tetraethylammonium. In correlation with the macroscopic currents there are single-channel gating modes ranging from the fast-inactivation mode which underlies the transient A-type current, to slow-inactivation modes characterized by bursts of longer openings, and corresponding to the slowly inactivating macroscopic currents.

Animals↗
PubMed1992

The sodium channel activator Brevetoxin-3 uncovers a multiplicity of different open states of the cardiac sodium channel.

The interaction of Brevetoxin 3 (Pbtx-3), a sodium channel activator, with the cardiac sodium channel was studied at the single channel level. It was found that Pbtx-3 (20 microM) shifted steady-state activation to negative potentials, without major effects on the time course of macroscopic activation or macroscopic currents decay, as calculated from averaged single-channel records. Single-channel open times were found to be prolonged. Under the influence of the toxin, sodium channel openings could be observed frequently even at maintained depolarisation. These openings occurred to at least nine different subconductance levels of the open state with smaller conductivities than the maximal one and differed in their open times. Current amplitudes of these open substates were found to cluster around certain amplitude values. Appearance of substates at maintained depolarisation was dependent on the transmembrane potential (Em): Substates with smaller conductivity appeared more frequently at lower Em values whereas at higher Em values substates with higher conductivity values dominated. Furthermore, it was demonstrated that appearance of substates did not result from incomplete recovery from inactivation. From these observations it was concluded that the open substates observed correspond to different conformational states of the channel's activation gates. Under physiological conditions, when the sodium channel opens directly from its closed state these 'incomplete'-open states of the cardiac sodium channel are obscured by fast gating transitions between the corresponding, electrically silent, preopen states. Thus, Pbtx-3 acts mainly via stabilisation of the channel's preopen and different open states. A classification of sodium channel modifiers, based on their interaction with different conformational states of the channel is suggested.

Animals↗
PubMed1992

Stereoselective interactions of (R)- and (S)-propafenone with the cardiac sodium channel.

The specific interactions of both (R)- and (S)-propafenone with the cardiac sodium channel were studied with patch clamp techniques in the whole-cell recording mode at reduced extracellular Na+ on guinea pig ventricular cells. Both (R)- and (S)-propafenone (10 microM) shifted the membrane potential required for half-maximal steady-state inactivation (E0.5) of the cardiac sodium channel to considerably more negative membrane potentials [E0.5 = -70.8 +/- 2.9 mV for controls vs. -85 +/- 3.1 mV for (R)-propafenone and -91.9 +/- 1.7 mV for (S)-propafenone]. (S)-Propafenone at a concentration of 10 microM is more effective in shifting the h infinity curve of the cardiac sodium channel. Recovery from inactivation of the cardiac sodium current is prolonged by orders of magnitude by both stereoenantiomeric forms [time constants were estimated to be 38 +/- 15 ms at -90 mV vs. 46.5 +/- 14.3 s for (R)-propafenone and 74.2 +/- 37.9 for (S)-propafenone]. Development of block occurs mainly through the inactivated channel conformation for both (R)- and (S)-propafenone. Development of block of inactivated cardiac sodium channels occurs with time constants of 15.9 +/- 3.9 s for (R)-propafenone and 19.7 +/- 7.3 s for (S)-propafenone at 10 microM. Action potential duration and possible stereoselective interaction with ion transport systems other than sodium channels may influence the block developed by either (R)- or (S)-propafenone at a given concentration and beating frequency indirectly through the membrane potential.(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗
PubMed1992

Kinetic modulation of guinea-pig cardiac L-type calcium channels by fendiline and reversal of the effects of Bay K 8644.

1. The modulation of L-type calcium channel current (ICa) by fendiline, a diphenylalkylamine type of calcium channel blocker was investigated on guinea-pig ventricular myocytes by use of the whole-cell patch-clamp technique. 2. Fendiline-induced block of ICa is accompanied by modulation of the channel kinetics in a complex manner. The time course of ICa inactivation is significantly faster and the channel availability (f infinity) curve is shifted considerably to more negative potentials by fendiline. These findings can be interpreted qualitatively in terms of a modulated receptor. 3. When the 1,4-dihydropyridine agonist (4R, 4S)-Bay K 8644 was added in presence of 30 microM fendiline a further reduction of ICa instead of the expected stimulatory effect was observed. 4. A similar 'paradoxical' inhibition of ICa was produced by the pure agonist enantiomer (4S)-Bay K 8644. Thus this novel effect of Bay K 8644 cannot be attributed to changes in affinity of the 1,4-dihydropyridine receptor site for (4R)-Bay K 8644 during fendiline action. 5. The IC50 for fendiline was reduced to 3.0 +/- 0.1 microM (control value: 17.0 +/- 2.4 microM) and the Hill slope in its presence was increased to 1.90 +/- 0.1 (control value: 1.39 +/- 0.23) by 1 microM (4R, 4S)-Bay K 8644. 6. (4R,4S)-Bay K 8644 caused the expected stimulation of ICa in the presence of verapamil, diltiazem and nifedipine, overcoming the inhibitory effect of these calcium channel blockers. 7. The 'paradoxical' inhibitory effect of the agonist Bay K 8644 can be explained in terms of an allosteric interaction between fendiline and the dihydropyridine agonist.

3-Pyridinecarboxylic acid, 1,4-dihydro-2,6-dimethy↗
PubMed1992

Protein kinase A reduces voltage-dependent Na+ current in Xenopus oocytes.

The voltage-dependent Na+ channel of the brain is a good substrate for phosphorylation by the cAMP-dependent protein kinase (protein kinase A, or PKA), but the physiological effects of PKA on Na+ channels are poorly documented. We studied modulation by PKA of voltage-dependent Na+ channels expressed in Xenopus oocytes injected with RNA coding for the alpha-subunit of the channel protein (rat brain type IIA and its variant VA200), using the two electrode voltage-clamp technique. Intracellularly injected cAMP or catalytic subunit of PKA, or extracellularly applied forskolin, inhibited the Na+ current by 20-30%. The effect of cAMP was attenuated by prior injection of PKA inhibitors. Injection of small doses of protein phosphatase 2A increased the Na+ current by 10%, whereas larger doses of protein phosphatase 1 and alkaline phosphatase were without effect. The inhibition by PKA showed little voltage dependence, being only slightly stronger at holding potentials at which the availability of the channels was reduced. The voltage dependence of activation and inactivation processes was not altered by cAMP. Similar effects were exerted by forskolin and cAMP on the Na+ channels expressed after the injection of heterologous (total) RNA from rat brain. Thus, PKA modulates the Na+ channel by a mechanism that does not involve major changes in the voltage dependency of the current and is exerted on the channel-forming alpha-subunit.

Animals↗
PubMed1991

Molecular mechanism of protein kinase C modulation of sodium channel alpha-subunits expressed in Xenopus oocytes.

The mechanism of modulation of sodium channel alpha-subunits (Type IIA) by a protein kinase C (PKC) activator was studied on single channel level. It was found that: (i) time constants for channel activation were prolonged; (ii) inactivation remained virtually unchanged; (iii) peak sodium inward current was reduced as evidenced by calculation of average sodium currents; and (iv) time constants for current activation and decay were prolonged. (i), (iii) and (iv) were voltage dependent, being most prominent at threshold potentials. The data show that a voltage dependent action on the activation gate can account for the observed reduction of peak inward sodium current and prolongation of current decay in macroscopic experiments.

Animals↗
PubMed1990

Inhibition of the fast sodium inward current in ventricular cardiomyocytes of rats and guinea pigs by a novel potent sodium channel blocking agent.

In enzymatically-dispersed single ventricular cardiomyocytes of adult rats and guinea pigs the inhibition of the cardiac sodium current by a novel sodium channel blocking agent (LG 83-6-05, 1-[3-(2-Hydroxy-3-(2-methylpropylamino)-propoxy)-4-methyl-2- thienyl++ ]-3-phenyl-1-propanon hydrochloride) was studied. A single-electrode voltage-clamp system (switch clamping, patch electrodes) was used to measure action potentials as well as ionic currents during voltage-clamp experiments. In addition single channel measurements were performed using the patch-clamp technique. The single cell system enabled us to demonstrate that LG 83-6-05 is an inhibitor of the cardiac sodium current. The substance acts concentration-dependently and belongs to the most potent sodium-channel blocking agents known. It could be shown that the whole-cell sodium inward current is blocked in a frequency-dependent manner (phasic block) and that the steady-state inactivation curve of the sodium current is shifted significantly towards negative potentials, indicating a considerable tonic block at the resting membrane potential. The time constant of the recovery from inactivation of the sodium current as estimated from voltage-clamp experiments is prolonged by a factor of up to 290 (holding potential -110 mV, 2 mumol/l). This prolongation is voltage dependent, faster release from block occurring at more negative potentials. The open state probability of the single cardiac sodium channel is reduced in a frequency-dependent manner, whereas its current amplitude remains unchanged during the influence of the substance. The number of channels not available for opening is increased considerably with increasing stimulus frequency. These findings suggest stabilization of the inactivated state of the ionic channel by drug binding.

Action Potentials↗
PubMed1989

The cardiac sodium channel shows a regular substate pattern indicating synchronized activity of several ion pathways instead of one.

Cardiac sodium channel substates were induced by using different gating modifiers, namely S-DPI 201-106 (s), toxin II from Anemonia sulcata (a), veratridine (v) and mixtures of these agents (s + v, a + v). Current ratios (normalized substate currents), slope conductances, reversal potentials and saturation characteristics were evaluated for the individual channel substates. The results can be summarized as follows: (i) Current ratios fell into a pattern of six equidistant values (I to VI) irrespective of the modification applied (0.20, 0.34, 0.51, 0.69, 0.85, 1.00). Slope conductances, determinable for substates II, V and VI (4.8, 11.7 and 14.0, respectively), are also consistent with six conductance substates which are integer multiples of a smallest conductance (state I). (ii) The permeability ratio PNa+/PK+ (i.e., reversal potential of substate currents) of the sodium channel was conserved both for different modifications, i.e., by s, a, s + v and a + v, and for the different substates (at least for II, IV and VI) observed for each modification. (iii) Sodium binding to the channel is substate independent. Analysis of slope conductances of states II and VI for three sodium chloride concentrations (71.5, 140 and 303 mM) revealed different maximal conductances (geVImax = 2.9.geIImax) but similar apparent affinities for sodium (KNa + VI = 286 mM; KNa + II = 303 mM). These findings are shown to seriously challenge the commonly unquestioned conception that 'single-current events' reflect ion passage through only one single pathway. The alternative view, that not one pore, but either six or three pores with synchronized gating ('oligochannel') underlie 'single-channel events', is shown to readily account for the observed substate properties and appears not to contradict known properties of 'the sodium channel'. This fundamentally new view of the sodium channel aims to invoke further efforts to distinguish between conceptually distinct models of structure-function relationships for a variety of channels which show multiple substates and conserved ion selectivity.

Animals↗
PubMed1988

Calcitonin gene-related peptide: release by capsaicin and prolongation of the action potential in the guinea-pig heart.

The mechanisms underlying the stimulatory effects of capsaicin on the contractility of the guinea-pig heart were studied in vitro. Capsaicin (10(-7) to 10(-5) M) caused an increased overflow of immunoreactive material, suggesting release of calcitonin gene-related peptide (CGRP)-and neurokinin A (NKA)-like immunoreactivity (-LI), but not of neuropeptide Y (NPY)-LI from the isolated Langendorff-perfused whole heart. The capsaicin-induced release was calcium-dependent. During exposure to capsaicin, the heart rate was increased, while the contractile force was reduced. In addition to releasing CGRP and NKA-LI, potassium (60 mM) also increased the overflow of NPY-LI. The potassium-induced release of peptides was less calcium-dependent than the response to capsaicin. Considerably higher tissue levels of CGRP-LI were found in the atria (about 30 pmol g-1) than in the ventricles (about 10 pmol g-1). In experiments on the right atria using transmembrane action-potential recordings of myocytes, CGRP induced a prolongation of the action potential concomitantly with an increase in rate and contractile force, which was similar to the effect of noradrenaline. Furthermore, CGRP increased the contractile force and relaxation velocity of the electrically stimulated atria. Capsaicin (10(-7) M) also increased the duration of the atrial action potential. In conclusion, CGRP-like material is released by capsaicin from the isolated guinea-pig heart. Both CGRP and capsaicin prolong the plateau phase of the action potential of atrial myocytes. Therefore, the present data give further evidence that CGRP release from sensory nerves within the heart underlies the cardiostimulatory actions of capsaicin.

Action Potentials↗
PubMed1987

A mechanistic interpretation of the action of toxin II from Anemonia sulcata on the cardiac sodium channel.

Cardiac sodium channels, modified by Anemonia sulcata toxin II, have been analyzed by the patch-clamp method. The open state of the modified sodium channels proved to be prolonged highly significantly and reopening from a closed state denoted c*-state frequently occurred, interrupted by silent periods, denoted i*-state. Activation from the c*-state was apparently not affected by toxin action, whereas activation from the i*-state was markedly prolonged. Upon higher depolarizations toxin-induced sodium channels disappeared and this behaviour has been attributed to dissociation of the toxin from the channel by use of a special pulse-protocol. The onset of the toxin effect on the action potential proved to depend on stimulation, and it is concluded that the toxin binds preferentially to the open (o)-state. Taking together the results, a kinetic scheme is suggested for action of the toxin on the cardiac sodium channel.

Action Potentials↗
PubMed1987

Analysis of single channel currents with a microprocessor based device.

Data evaluation of single channel currents obtained from artificial black lipid membranes and with the patch clamp method is an important part of every single channel study, but it is a time consuming part often exceeding the time for experimentation and recording by far. We describe here a microprocessor based device, which allows the experimentator to analyse in a simple way the distribution of current levels in a single channel trace (amplitude-histogram analysis of single channel currents) either online, or offline. Current levels are sampled at a constant frequency of 6 kHz and the relative frequencies of occurrence of these current levels are displayed as a histogram on the screen of an analog or digital storage oscilloscope. The data reducing algorithm of this analyser eliminates the requirement of large amounts of mass storage that normally is needed for digital amplitude-histogram analysis of single channel recordings. Examples of evaluation for both a voltage operated cation-channel and a blockage of a potassium channel by tetraethylammoniumchloride (TEA) are given.

Animals↗
PubMed1985

Single voltage-dependent and outward rectifying K+-channels in isolated rat heart cells.

Studies on single K+-channel currents recorded from isolated rat heart muscle cells, in which early repolarization is known to be exceptionally fast, are reported here. A K+-channel which is blocked by TEA (tetraethylammonium) from the inside only has been found. The total open time of the channel, measured in steady-state after activation, indicated outward rectifying properties. The single channel conductance increases with depolarization from 25 pS at -70 mV to 75 pS at + 70 mV. Selectivity of the channel has also been measured and it was found that only Rb+ and K+ can permeate the channel, whereas the permeability (P) for Li+, Na+, Cl-, Mg2+, and Ca2+ is less than 0.05 times PK+. Ba2+ and CS+ block the channel activity. These results clearly demonstrate the existence of K+-selective outward rectifying conductance pathways in rat ventricular myocytes.

Animals↗