Potassium current

Both erythromycin and clarithromycin have been reported to cause QT prolongation and the cardiac arrhythmia torsade de pointes in humans, however direct evidence documenting that these drugs produce this effect by blocking human cardiac ion channels is lacking. The goal of this study was to test the hypothesis that these macrolide antibiotics significantly block the delayed rectifier current (IKr) encoded by HERG (the human ether-a-go-go-related gene) at drug concentrations, temperature and ionic conditions mimicking those occurring in human subjects. Potassium currents in HEK 293 cells stably transfected with HERG were recorded using a whole cell voltage clamp method. Exposure of cells to erythromycin reduced the HERG encoded potassium current in a concentration dependent manner with an IC50 of 38.9 +/- 1.2 microM and Hill Slope factor of 0.4 +/- 0.1. Clarithromycin produced a similar concentration-dependent block with an IC50 of 45.7 +/- 1.1 microM and Hill Slope factor of 1.0 +/-...

Mathematical models play a crucial role in the deciphering of complex systems both before and after a clear hypothesis exists for the internal mechanism of the system. For example. when little or nothing is known about the internal processes of a system. it may be possible to summarize the results of a large number and diversity of experiments in a few lines of mathematics by fitting the results with carefully chosen equations. This approach is usually pursued in the hope that the data will be accurately fitted by a familiar set of equations and that they in turn will suggest a possible internal mechanism. A well known example of this form of mathematical modelling in ion transport is the analvsis bv Hodgkin and Huxlev (1952) of the voltage clamp records from the squid giant axon . Resolution of the currents into components due to specific ions was possible through a variety of experiments. but it was not
possible to be specific about a physico-chemical mechanism as a basis for a kinetic analysis of the individual ion conductances. Hodgkin and Huxley chose equations that would represent the data with reasonable accuracy and that would be simple enough to facilitate further theoretical calculations.

SUMMARY AND CONCLUSIONS 1. We study the propagation and dynamics of spindle waves in thal amic slices by developing and analyzing a model of recipro-cally coupled populations of excitatory thalamocortical (TC) neu-rons and inhibitory thalamic reticular (RE) neurons. ...

There has been an increasing interest in compounds that modulate potassium ion channels (K + -channels) since they can be developed as important therapeutic agents against ischemic heart diseases. Of the diverse family of K + - channels, the voltage-gated potassium channel Kv1.3 constitutes an attractive target for the selective suppression of effec- tor memory T cells in autoimmune diseases.

We present a new algorithm (SATCHMO) that simultaneously estimates a tree and generates a set of multiple sequence alignments given a set of protein sequences. Alignments are constructed for each node in the tree. These alignments predict the structurally conserved elements of the sequences in a subtree and are therefore of different lengths, and represent different amino acid preferences, at different nodes. Hidden Markov Models (HMMs) are also generated for each node and are used to determine branching order, to align sequences and to predict structurally alignable regions. In experiments on the BAliBASE benchmark alignment database, SATCHMO is shown to perform comparably to ClustalW and the UCSC SAM HMM software. Results using SATCHMO to identify protein domains are demonstrated on potassium channels, with implications for the mechanism by which tumor necrosis factor alpha affects potassium current.

Background Periventricular leukomalacia (PVL) is a frequent complication of preterm delivery. Proinflammatory cytokines, such as interferon-γ (IFN-γ) and tumor necrosis factor α (TNF-α) released from astrocytes and microglia activated by infection or ischemia have previously been shown to impair survival and maturation of oligodendrocyte progenitors and could thus be considered as potential factors contributing to the generation of this disease. The first goal of the present study was to investigate whether exposure of oligodendrocyte precursors to these cytokines arrests the maturation of ion currents in parallel to its effects on myelin proteins and morphological maturation. Secondly, in the search for agents, that can protect differentiating oligodendrocyte precursor cells from cytokine-induced damage we investigated effects of coapplications of corticosteroids with proinflammatory cytokines on the subsequent survival and differentiation of oligodendrocyte progenitor cells. Metho...

The principal cells of the chick tangential nucleus are second-order sensory neurons that participate in the three-neuron vestibulo-ocular and vestibulocollic reflexes. In postnatal animals, second-order vestibular neurons fire repetitively on depolarization. Previous studies have shown that, although this is an important feature for normal reflex function, it is only acquired gradually during embryonic development. Whereas at 13 embryonic days (E13) the principal cells accommodate after firing a single spike, at E16 a few principal cells repetitively can fire multiple action potentials on depolarization. Finally, in the hatchling, the vast majority of principal cells is capable of nonaccommodating firing on depolarization. As a first step in understanding the mechanisms underlying developmental change in excitability of these second-order vestibular neurons, we analyzed the outward potassium currents and their role in accommodation, using brainstem slices at E16. The principal cell...

Rat superior cervical ganglion (SCG) neurons express low- threshold noninactivating M-type potassium channels (IK(M)), which can be inhibited by activation of M1 muscarinic recep- tors. This inhibition occurs via pertussis toxin-insensitive G-proteins belonging to the Gaq family (Caulfield et al., 1994). We have used DNA plasmids encoding antisense sequences against the 39 untranslated regions of Ga subunits (antisense plasmids) to

Rat superior cervical ganglion (SCG) neurons express lowthreshold noninactivating M-type potassium channels (I K(M)), which can be inhibited by activation of M1 muscarinic receptors. This inhibition occurs via pertussis toxin-insensitive G-proteins belonging to the G � q family (Caulfield et al., 1994). We have used DNA plasmids encoding antisense sequences against the 3 � untranslated regions of G � subunits (antisense plasmids) to investigate the specific G-protein subunits involved in muscarinic inhibition of I K(M). These antisense plasmids specifically reduced levels of the target G-protein 48 hr after intranuclear injection. In cells depleted of G � q, muscarinic inhibition of I K(M) was attenuated compared both with uninjected neurons and with neurons injected with an inappropriate G � oA antisense plasmid. In contrast, depletion of G � 11 protein did not alter I K(M) inhibition. To determine whether the � or ��

Transient potassium currents distinctively affect firing properties, particularly in regulating the latency before repetitive firing. Pyramidal cells of the dorsal cochlear nucleus (DCN) have two transient potassium currents, I Kif and I Kis, fast and slowly inactivating, respectively, and they exhibit firing patterns with dramatically variable latencies. They show immediate repetitive firing, or only after a long latency with or

Gamma (30-80 Hz) and beta (12-30 Hz) oscillations such as those displayed by in vitro hippocampal (CA1) slice preparations and by in vivo neocortical EEGs often occur successively, with a spontaneous transition between them. In the gamma rhythm, pyramidal cells fire together with the interneurons, while in the beta rhythm, pyramidal cells fire on a subset of cycles of the interneurons. It is shown that gamma and beta rhythms have different properties with respect to creation of cell assemblies. In the presence of heterogeneous inputs to the pyramidal cells, the gamma rhythm creates an assembly of firing pyramidal cells from cells whose drive exceeds a threshold. During the gamma to beta transition, a slow outward potassium current is activated, and as a result the cell assembly vanishes. The slow currents make each of the pyramidal cells fire with a beta rhythm, but the field potential of the network still displays a gamma rhythm. Hebbian changes of connections among the pyramidal c...

The Kv4 subfamily of voltage-gated potassium channels is responsible for the transient A-type potassium current that operates at subthreshold membrane potentials to control membrane excitability. Arachidonic acid was shown recently to modulate both the peak amplitude and kinetics of the hippocampal A-current. However, in Xenopus oocytes, arachidonic acid only inhibited the peak amplitude of Kv4 current without modifying its kinetics. These results suggest the existence of Kv4 auxiliary subunit(s) in native cells. We report here a K-channel interacting protein (KChIP)-dependent kinetic modulation of Kv4.2 current in Chinese hamster ovary cells and Kv4.2 and Kv4.3 currents in Xenopus oocytes by arachidonic acid at physiological concentrations. This concentration-dependent effect of arachidonic acid resembled that observed in cerebellar granule neurons and was fully reversible. Other fatty acids, including a nonhydrolyzable inhibitor of both lipooxygenase and cyclooxygenase, 5,8,11,14-...

There are two distinct voltage-dependent K+ currents in muscle fibers freshly isolated from the human flatworm parasite S. mansoni. Present is a delayed rectifier current with a tau act of 17 msec and tau inact > 3 sec. The delayed rectifier is very resistant to steady-state inactivation, with over 40% of the current non-inactivating, and over 15 sec required for the maximum inactivation of the other portion. The current is resistant to block by extracellular tetraethylammonium, is half-blocked by 10 mM 4-aminopyridine, and is insensitive to dendrotoxin. Also present is an "A" current with tau act < 1 msec and tau inact < 15 msec. The "A" current, like the delayed rectifier current, is resistant to block by external tetraethylammonium and is insensitive to dendrotoxin. Three micromoles of 4-aminopyridine produce a half-blockade of the "A" current. These two K+ currents are very similar to a delayed rectifier and "A" currents that have...

A slow muscarinic EPSP, accompanied by an increase in membrane input resistance, can be elicited in hippocampal CA1 pyramidal cells in vitro by electrical stimulation of cholinergic afferents in the slice preparation. Associated with the slow EPSP is a blockade of calcium-activated potassium afterhyperpolarizations (AHPs) (Cole and Nicoll, 1984a). In this study a single-electrode voltage clamp was used to examine the currents affected by activation of muscarinic receptors, using either bath application of carbachol or electrical stimulation of the cholinergic afferents. The 3 main findings of this study are that (1) of the 2 calcium-activated potassium currents (termed IAHP and IC) in hippocampal pyramidal cells, only IAHP is sensitive to carbachol; (2) IAHP is approximately 10-fold more sensitive to carbachol than is another muscarine-sensitive current, IM; and (3) neither blockade of IAHP nor of IM can account for the production of the slow EPSP. Rather, the slow EPSP appears to b...

BK channels modulate cell firing in excitable cells in a voltage-dependent manner regulated by fluctuations in free cytosolic Ca(2+) during action potentials. Indeed, Ca(2+)-independent BK channel activity has ordinarily been considered not relevant for the physiological behaviour of excitable cells. We employed the patch-clamp technique and selective BK channel blockers to record K(+) currents from bovine chromaffin cells at minimal intracellular (about 10 nM) and extracellular (free Ca(2+)) Ca(2+) concentrations. Despite their low open probability under these conditions (V(50) of +146.8 mV), BK channels were responsible for more than 25% of the total K(+) efflux during the first millisecond of a step depolarisation to +20 mV. Moreover, BK channels activated about 30% faster (τ = 0.55 ms) than the rest of available K(+) channels. The other main source of fast voltage-dependent K(+) efflux at such a low Ca(2+) was a transient K(+) (I(A)-type) current activating with V (50) = -14.2 m...

We investigated the effects of H 2 O 2-induced oxidative stress on the delayed-rectifier current (IK DR), neuronal physiological and morphological properties. Measurements were obtained from hippocampal CA1 neurons in control solution and from ...

Whole-cell voltage-clamp techniques were used to study the effects of the protein kinase C (PKC) activators phorbol esters and OAG on Ca and K currents in differentiated neurons acutely dissociated from adult hippocampus and in tissue-cultured neurons from fetal hippocampus. PKC activators had selective depressant effects on K currents, with persistent currents (IK and IK-Ca) being reduced and transient current (IA) being unaffected. In both cell types we recorded both high-voltage-activated, noninactivating (L-type) and high-voltage-activated, rapidly inactivating (N-type) Ca current. A low-voltage-activated, rapidly inactivating (T-type) Ca current was also recorded in tissue-cultured neurons but not in acutely dissociated neurons. PKC activators markedly reduced N-type current with less effect on L-type and no effect on T-type Ca current. Effects of PKC activators could be reversed with washing or with application of PKC inhibitors H-7 or polymyxin-B, an effect that could not be ...