Slow Potassium Channel Research Articles

Alterations of gating parameters by neutral substitutions of transmembrane Leu52 of slow potassium channel.

Alterations of gating parameters by neutral substitutions of transmembrane Leu52 of slow potassium channel.

Oxytocin actions on voltage-dependent ionic channels in pregnant rat uterine smooth muscle cells.

The effects of oxytocin, a uterotonic polypeptide hormone, on the voltage-dependent slow calcium, fast sodium, and potassium channel currents were studied using whole-cell voltage clamp of freshly isolated cells from late pregnant (18-21 day) rat myometrium. The calcium current was rapidly inhibited by oxytocin (about 25% inhibition at 20 nM) in a dose-dependent manner, and this inhibitory effect was completely reversible by washout. However, inhibition was not observed when barium was used as the charge carrier. Sodium current and potassium current were not modified by oxytocin, thus sodium and potassium currents may not play important roles in oxytocin-induced augmentation of uterine contraction. It is concluded that oxytocin stimulates uterine contraction by mechanisms other than augmentation of the voltage-dependent calcium current, e.g., by release of Ca from sarcoplasmic reticulum (by inositol triphosphate) or by activation of a receptor-operated Ca channel. The inhibition of the slow calcium current may be induced by the elevation of [Ca]i.

Alteration of channel activities and gating by mutations of slow ISK potassium channel.

ISK is a small membrane protein consisting of 129-130 amino acid residues with a single putative transmembrane domain and induces a very slow voltage-dependent K+ channel activity in the Xenopus oocyte expression system. We investigated the nature and structure-function relation of ISK by examining the effects of various mutations of ISK on the K+ channel activities measured in Xenopus oocytes. Deletion and truncation of the ISK protein indicated that the 63-amino acid sequence covering a transmembrane domain is sufficient for eliciting a K+ channel activity characteristic of ISK. Amino acid substitutions at a total of 31 positions within and surrounding the transmembrane domain caused different effects on the channel activity. A channel activity was enhanced by substitution of leucine with isoleucine at position 52 within the transmembrane domain, and the kinetic analysis of this mutation indicated that the enhancement of the channel activity is due to an alteration of a gating property of the ISK protein and thus supported the view that ISK forms an integral part of the K+ channel itself. The substitutions at many positions of the membrane-following region produced drastic reduction of the channel activity, and this is in marked contrast to the lack of effects of amino acid substitutions at the membrane-preceding region. Thus, the cytoplasmic portion immediately following the transmembrane domain plays a crucial role in inducing the channel activity of ISK.

Effects of a slow potassium permeability on repetitive activity of the frog node of ranvier

Adding a potassium permeability with slow kinetics to the Frankenhaeuser-Huxley equations describing action potential generation at a frog node of Ranvier has a twofold effect on the maintained repetitive firing the model can show. If the contribution of the slow to the total potassium permeability is increased, the maintained discharge frequency for a given stimulating current experiences a decrease. On the other hand, addition of the slow channel narrows the range of currents for which the model can generate repetitive activity. If as little as 6.2% of the total potassium permeability are provided by the slow channels, the Frankenhaeuser-Huxley equations completely lose the ability to show maintained firing. The introduction of the slow potassium current abolishes especially repetitive activity at low values of stimulating current. This effect is so marked that the minimal discharge frequency the model can maintain increases with increasing contribution of the slow channel. Therefore, an important purpose of the slow potassium channel present at the frog nodal membrane could consist of preventing the node of Ranvier from generating consistent firing on its own.

Heterogeneous distribution of fast and slow potassium channels in myelinated rat nerve fibres.

1. Potassium currents were measured in voltage-clamped single myelinated rat nerve fibres before and after paranodal demyelination with 0.2% pronase or 0.2% lysolecithin added to the external solution. Sodium currents were blocked by 300 nM-tetrodotoxin. For the purpose of comparison, intact frog nerve fibres were also investigated. 2. Our results suggest the existence of at least two distinct types of K+ channels in the intact node of Ranvier, one with slow and another with fast gating kinetics, in the ratio 4:1. 3. In the rat nodal membrane, slow K+ channels have voltage-dependent time constants of K+ deactivation with tau n = 68 ms at E = -105 mV and tau n = 26 ms at E = -150 mV at 20 degrees C. The activation curve of the slow K+ conductance is sigmoid with an inflexion point at -60 mV. This means that about 35% of the slow K+ channels are in the open state at the resting potential of -77 mV. Slow K+ channels could be blocked by 10 mM-tetraethylammonium chloride, but were insensitive to 4-aminopyridine. 4. After paranodal demyelination the ratio of fast to slow K+ channels increased from 17 to 83%. As in the frog (Dubois, 1981 alpha), the population of fast K+ channels in the rat may consist of two different subgroups, both of which can be blocked by 4-aminopyridine. 5. Demyelination was accompanied by an increase in the capacity current which was used to estimate the exposed membrane area. The density of slow and fast K+ channels was calculated from the quotient of the steady-state K+ conductance to membrane area. The density of the slow K+ channels is maximal in the nodal membrane and decreases to 1/31 in the internode. By contrast, the distribution of the fast K+ channels differs, their density being maximal in the paranode and decreasing to one-sixth in the node and internode.

Depolarization changes the mechanism of accommodation in rat and human motor axons.

1. We have previously studied accommodation in rat and human motor axons by testing excitability with combinations of long and short current pulses. We found that normally polarized axons accommodate slowly and partially (over about 50 ms) to subthreshold depolarizing currents, and that the principal mechanism is the activation of slow potassium channels (Bostock & Baker, 1988). To understand the response of human nerves to ischaemia, we have now extended these observations to axons already depolarized before the testing currents were applied. 2. Rat ventral root axons were depolarized by passing continuous currents or by raising the extracellular potassium concentration. Human forearm nerves were depolarized by ischaemia, induced by inflating a sphygmomanometer cuff on the upper arm. Depolarized rat and human motor axons accommodated much more rapidly and completely than normally polarized axons (e.g. accommodation in rat axons was 50% complete within 2 ms at about 15 mV depolarized to rest). 3. The fast component of accommodation in depolarized rat fibres was not blocked by tetraethylammonium ions or 4-aminopyridine, was not accompanied by a conductance or potential change, and had a time constant of 1.7 ms at 30 degrees C. It was attributed to inactivation of closed sodium channels. 4. In depolarized rat fibres exhibiting fast accommodation, a brief rise in excitability was seen at the break of an anodal current. Our prediction that human motor axons would show anode-break excitation during ischaemia was readily confirmed. 5. The results are discussed in relation to Hill's (1936) mathematical description of accommodation in nerve, and it is concluded that his description is only applicable to depolarized axons.

Patch-clamp studies of slow potential-sensitive potassium channels in longitudinal smooth muscle cells of rabbit jejunum.

1. The patch-clamp technique was used to study single channel currents in membrane patches of longitudinal smooth muscle cells of rabbit jejunum dispersed by collagenase treatment. Recordings were made from both cell-attached and isolated patches.2. The predominant unit currents observed were outward at membrane potentials positive to the potassium equilibrium potential (E(K)) and they were rapidly and reversibly blocked by tetraethylammonium (TEA). Their size varied as E(K) was changed but was not noticeably affected by changing E(Na), E(Cl) or E(Ca); it was little altered in calcium-free EGTA solution. Thus, these currents apparently result mainly, if not exclusively, from the movements of potassium ions through channels insensitive to the calcium ion concentration. The present study describes the properties of these potassium channels.3. The unit conductance varied slightly with potential in most experiments; around zero potential it was about 50 pS. The conductance was dependent upon the potassium, but not the calcium, gradient. Sub levels of conductance of about two-thirds and, less commonly, one-third of the fully conducting channel state were sometimes seen.4. Membrane patches were studied which showed one to about twelve levels of outward current which were presumed to result from the opening of up to twelve channels having the same characteristics. The probability of channel open state varied with membrane potential, increasing in the potential range -40 to +40 mV. Channel openings were rare negative to -40 mV. No inward currents through these potassium channels were observed as openings were not seen at membrane potentials negative to E(K).5. When the probability of channel opening was low, channel openings occurred in bursts which could be separated by several seconds. Analysis of the openings of a single channel revealed that open times and short closed times were exponentially distributed with mean durations of 15-45 ms and about 6 ms at zero potential. In some patches regular cyclical openings of several channels occurred. In other patches openings of individual channels appeared to be independent events as they were reasonably fitted by a binomial distribution.6. Following a step change from negative potentials, where channels were closed, to more positive potentials, channel openings increased during a period of 10 s to reach a steady state. No evidence of inactivation was observed.7. These results suggest the existence of a population of potential-sensitive potassium-selective ion channels in the smooth muscle cell membrane which are closed at the resting membrane potential and which open upon depolarization with slow (seconds) kinetics; these may be involved in the slow potential (wave) activity of this muscle.

The Cole-Moore effect in nodal membrane of the frog Rana ridibunda: evidence for fast and slow potassium channels.

The K conductance (gK) kinetics were studied in voltage-clamped frog nodes (Rana ridibunda) in double-pulse experiments. The Cole-Moore translation for gK--t curves associated with different initial potentials (E) was only observed with a small percentage of fibers. The absence of the translation was found to be caused by the involvement of an additional, slow, gK component. This component cannot be attributed to a multiple-state performance of the k channel. It can only be accounted for by a separate, slow K channel, the fast channel being the same as the n4 K channel in R. pipiens. The slow K channel is characterized by weaker sensitivity to TEA, smaller density, weaker potential (E) dependence, and somewhat more negative E range of activation than the fast K channel. According to characteristics of the slow K system, three types of fibers were found. In Type I fibers (most numerous) the slow K channel behaves as and n4 HH channel. In Type II fibers (the second largest group found) the slow K channel obeys the HH kinetics within a certain E range only; beyond this range the exponential decline of the slow gK component is preceded by an E-dependent delay, its kinetics after the delay being the same as those in Type I fibers. In Type III fibers (rare) the slow K channel is lacking, and it is only in these fibers that the Cole-Moore translation of the measured gK--t curves can be observed directly. The physiological role of the fast and slow K channel in amphibian nerves is briefly discussed.

The spatial distribution of ionic conductances in normal and axotomized motorneurons

In a previous paper, a model was developed to study and contrast the relations between cell geometry and repetitive firing in large and small motorneurons. This model is extended and revised in the present paper and used to analyze certain experimental findings in normal and axotomized motorneurons. (1) For normal motorneurons, in order to reproduce the experimentally observed difference in interspike voltage trajectory (concave vs convex) in the primary and secondary ranges respectively of the current-firing rate curve, it is necessary to localize the major part of a slow potassium conductance on the motorneuron dendrites. This slow potassium conductance leads to the afterhyperpolarization following an action potential. (2) In order to activate the slow potassium conductance after an action potential, we have postulated a calcium conductance in the motorneuron membrane, as suggested by the work of Barrett and Barrett, and Krnjevid and Lisiewicz. In normal motorneurons, the proximal dendrites must display a certain amount of sodium conductance (g Na) in order to activate the calcium conductance (g Ca) sufficiently to ‘trigger’ the slow potassium channel. (3) With the above mechanism for calcium and slow potassium conductances, the model shows that the firing rate in the primary range is mainly determined by the decay of the slow potassium conductance which generates the long afterhyperpolarization. In the secondary range, this slow potassium conductance is essentially constant between spikes, so that the firing rate is determined by fast potassium recovery. (4) The model is further able to display several features of the partial responses seen in axotomized motorneurons: for example, their all-or-nothing character following different levels of synaptic excitation, and the hump on the trailing part of an action potential. The model achieves this when a patch of membrane with an increased sodium and fast potassium conductance (a ‘hot spot’) is localized on a dendritic branch about 0.4–0.5 or more space constants away from the soma. (5) For an axotomized motorneuron, we were able to obtain simultaneously the correct current-firing rate curve (a single, high-slope region), and also the experimentally observed convex interspike trajectories, by manipulating slow potassium and ‘hot spot’ g Na parameters; this could be done by moving the ‘hot spots’ distally (to 0.6–0.7 space constants from the somas With a ‘hot spot’ close to the soma, shifting the Hodgkin-Huxley rate functions α m and α h (which lowers the voltage threshold) tended to accentuate the amount of primary range present; this was not observed with ‘hot spots’ located more distally. (6) An important variable in determining the nature of repetitive firing in both normal and axotomized motorneurons is the height of the dendritic potential (in the region in which the afterhyperpolarization is developed) immediately following the soma action potential.