Voltage Of Half-maximal Inactivation Research Articles

Effects of neomycin on high-threshold Ca 2+ currents and tetrodotoxin-resistant Na + currents in rat dorsal root ganglion neuron

High-threshold Ca 2+ channels and tetrodotoxin-resistant Na + channels are highly expressed in small dorsal root ganglion neurons. In acutely isolated rat dorsal root ganglion neurons, the effects of neomycin, one of the aminoglycoside antibiotics, on high-threshold Ca 2+ currents and tetrodotoxin-resistant Na + currents were examined using whole-cell patch recording. We showed for the first time that neomycin dose-dependently inhibited peak high-threshold Ca 2+ currents and peak tetrodotoxin-resistant Na + currents with half-maximal inhibitory concentrations at 3.69 μM ( n=20) and 1213.44 μM ( n=25), respectively. Inactivation properties of high-threshold Ca 2+ currents and activation properties of tetrodotoxin-resistant Na + currents were also affected by neomycin with reduction of excitability of small dorsal root ganglion neurons. Half-maximal inactivation voltage of high-threshold Ca 2+ currents was −45.56 mV before and −50.46 mV after application of neomycin ( n=10). Half-maximal activation voltage of tetrodotoxin-resistant Na + currents was −19.93 mV before and −11.19 mV after administration of neomycin ( n=15). These results suggest that neomycin can inhibit high-threshold Ca 2+ currents and tetrodotoxin-resistant Na + currents in small dorsal root ganglion neurons, which may contribute to neomycin-induced peripheral and central analgesia.

HKChIP2 is a functional modifier of hKv4.3 potassium channels: cloning and expression of a short hKChIP2 splice variant.

The Ca(2+) independent transient outward K(+) current (I(to1)) in the heart is responsible for the initial phase of repolarization. The hKv4.3 K(+) channel alpha-subunit contributes to the I(to1) current in many regions of the human heart. Consistently, downregulation of hKv4.3 transcripts in heart failure and atrial fibrillation is linked to reduction in I(to1) conductance. The recently cloned KChIP family of calcium sensors has been shown to modulate A-type potassium channels of the Kv4 K(+) channel subfamily. We describe the cloning and tissue distribution of hKChIP2, as well as its functional interaction with hKv4.3 after expression in Xenopus oocytes. Furthermore, we isolated a short splice variant of the hKChIP2 gene (hKCNIP2), which represents the major hKChIP2 transcript. Northern blot analyses revealed that hKChIP2 is expressed in the human heart and occurs in the adult atria and ventricles but not in the fetal heart. Upon coexpression with hKv4.3 both hKChIP2 isoforms increased the current amplitude, slowed the inactivation and increased the recovery from inactivation of hKv4.3 currents. For the first time we analyzed the influence of a KChIP protein on the voltage of half-maximal inactivation of Kv4 channels. We demonstrate that the hKChIP2 isoforms shifted the half-maximal inactivation to more positive potentials, but to a different extent. By elucidating the genomic structure, we provide important information for future analysis of the hKCNIP2 gene in candidate disorders. In the course of this work we mapped the hKCNIP2 gene to chromosome 10q24. Heteromeric hKv4.3/hKChIP2 currents more closely resemble native epicardial I(to1), suggesting that hKChIP2 is a true beta-subunit of human cardiac I(to1). As a result hKChIP2 might play a role in cardiac diseases, where a contribution of I(to1) has been shown.

Serotonin Enhances Central Olfactory Neuron Responses to Female Sex Pheromone in the Male Sphinx MothManduca sexta

In the brain of the sphinx moth Manduca sexta, sex-pheromonal information is processed in a prominent male-specific area of the antennal lobe called the macroglomerular complex (MGC). Whole-cell patch-clamp recordings from identified projection (output) neurons in the MGC have shown that serotonin [5-hydroxytryptamine (5-HT)] increases both the excitability of MGC projection neurons and their responses to stimulation with pheromone. At least two types of voltage-activated potassium currents in these cells are modulated by 5-HT. 5-HT decreases the maximal conductance of a transient potassium current (I(A)) and shifts its voltage for half-maximal inactivation to more negative potentials without affecting the half-maximal voltage for activation. This reduces the "window current" between the voltage activation and inactivation curves, decreasing the tonically active I(A) near the resting potential and causing the cell to depolarize. 5-HT's effect in this case is to decrease both the transient and resting K(+) conductance by modulating the same channel (I(A)). 5-HT also decreases the maximal conductance of a sustained potassium current [I(K(V))] without affecting its voltage dependence. Using HPLC, we show also that levels of 5-HT in the antennal lobes fluctuate significantly over a 24 hr period. Interestingly, 5-HT levels are highest at times when the moths are most active. We suggest that by controlling the responsiveness of antennal-lobe projection neurons to olfactory stimuli, 5-HT will have significant impact on the performance of odor-dependent behaviors.

Voltage-activated whole-cell K+ currents in lamina cells of the desert locust schistocerca gregaria

Voltage-dependent outward currents were studied in freshly dissociated somata of locust lamina cells. These currents were recorded in 142 somata using the whole-cell patch-clamp technique. By measuring the reversal potential at altered external [K+] and by replacing internal K+ with Cs+, we determined that the outward currents were carried by K+. The outward currents consist of a transient A-type K+ current (KA) and a delayed-rectifier-like K+ current (KD). Amongst the cells studied, we observed two distinct groups of cells. The most obvious difference between the two groups is that in group I cells the total outward current is dominated by KA (KA/KD=12.5), whereas in group II cells KA makes a smaller contribution (KA/KD=2.1). Furthermore, in cells of group I, the KA current shows a steeper voltage-dependence of activation, where VG50 is -29.9 mV and s is 11.9 (N=22), and inactivation, where VI50 is -84.5 mV and s is -6.3 (N=18), compared with the KA current in cells of group II: VG50=-7.9 mV; s=26.6 (N=36) and VI50=-68.4 mV; s=-7.5 (N=21) (VG50 is the voltage at which the whole-cell conductance G is half-maximally activated, VI50 is the voltage of half-maximal inactivation and s is the slope of the voltage-dependence). The transient KA current in group I cells decayed mono-exponentially. The decay of the KA current in group II cells was fitted with a double-exponential curve and was significantly faster than in group I cells. In contrast to the large differences in KA currents, the KD currents appeared to be quite similar in the two groups of cells.

An intermediate conductance K + channel in the cell membrane of mouse intestinal smooth muscle

Single channel currents were recorded from cell-attached and inside-out patches in smooth muscle cells of the mouse ileum in order to identify TEA-sensitive Ca 2+-dependent K + channels. Cells were bathed in high-K + (150 mM) solution with [Ca 2+] buffered to 80–150 nM with EGTA and patch pipettes were filled with low-K + (2.5 mM) physiological solution. Two distinct TEA-sensitive unitary outward current levels were identified at a holding potential ( V h) of 0 mV, corresponding to intermediate conductance (IK, ∼40 pS) and large conductance (BK, >200 pS) K + channels. The open probability ( P o) of IK channels increased with depolarization, the voltage for half-maximal activation averaging +12 mV in 80 nM Ca bath 2+. Raising the [Ca 2+] in the high-K + solution from 80 nM to 150 nM increased the P o of IK channels at V h=0 mV from 0.078 to 0.21. Likewise, the open probability of BK channels at 0 mV was increased from 0.003 to 0.026. Unlike BK channels, IK channels inactivated with maintained depolarization with a voltage for half-maximal inactivation of −66 mV. IK channels were blocked by 2–5 mM external TEA and were sensitive to both charybdotoxin (100 nM) and apamin (500 nM). Our results suggest that IK channels contribute significantly to the Ca 2+-dependent K + conductance in visceral smooth muscle.

Modulation of Cardiac Sodium Current by α1-stimulation and Volatile Anesthetics

Alpha1-adrenoceptor stimulation is known to produce electrophysiologic changes in cardiac tissues, which may involve modulations of the fast inward Na+ current (I(Na)). A direct prodysrhythmic alpha1-mediated interaction between catecholamines and halothane has been demonstrated, supporting the hypothesis that generation of halothane-epinephrine dysrhythmias may involve slowed conduction, leading to reentry. In this study, we examined the effects of a selective alpha1-adrenergic receptor agonist, methoxamine, on cardiac I(Na) in the absence and presence of equianesthetic concentrations of halothane and isoflurane in single ventricular myocytes from adult guinea pig hearts. I(Na) was recorded using the standard whole-cell configuration of the patch-clamp technique. Voltage clamp protocols initiated from two different holding potentials (V(H)) were applied to examine state-dependent effects of methoxamine in the presence of anesthetics. Steady state activation and inactivation and recovery from inactivation were characterized using standard protocols. Methoxamine decreased I(Na) in a concentration- and voltage-dependent manner, being more potent at the depolarized V(H). Halothane and isoflurane interacted synergistically with methoxamine to suppress I(Na) near the physiologic cardiac resting potential of -80 mV. The effect of methoxamine with anesthetics appeared to be additive when using a V(H) of -110 mV, a potential where no Na+ channels are in the inactivated state. Methoxamine in the absence and presence of anesthetics significantly shifted the half maximal inactivation voltage in the hyperpolarizing direction but had no effect on steady-state activation. The present results show that methoxamine (alpha1-adrenergic stimulation) decreases cardiac Na+ current in a concentration- and voltage-dependent manner. Further, a form of synergistic interaction between methoxamine and inhalational anesthetics, halothane and isoflurane, was observed. This interaction appears to depend on the fraction of Na+ channels in the inactivated state. (Key words: Anesthetics, volatile: halothane; isoflurane; methoxamine. Patch clamp: whole-cell configuration; sodium current; ventricular guinea pig myocytes.)

On the sodium and potassium currents of a human neuroblastoma cell line.

1. The patch-clamp method was applied to the study of ionic currents activated by depolarization of undifferentiated IMR-32 human neuroblastoma cells. Whole-cell sodium and potassium currents and single potassium ion channel currents from cell-attached patches were investigated. 2. Cells had a mean resting potential of -38 mV and mean input resistance of 1.6 G omega. Single action potentials were evoked under current clamp during the injection of depolarizing currents. 3. A voltage-dependent inward sodium current was observed which reversed at +44 mV. A Boltzmann fit to the activation curve gave a half-maximal activation voltage of -41.6 mV and a 'slope' of 3.9 mV. The steady-state inactivation curve had a half-maximal inactivation voltage of -81 mV and a 'slope' of 9.7 mV. 4. The time-dependent activation and inactivation of the current displayed classical Hodgkin-Huxley kinetics. Values for the time constants tau m and tau h of 0.16 and 0.63 ms were calculated for a voltage jump from -80 to -10 mV; tau m and tau h decreased as the step potential was changed from -30 to +20 mV. 5. Outward currents were activated in bathing solutions substantially free of anions and could thus be attributed to potassium ions. The tail current reversed in direction on repolarization to -60 mV when the potassium concentration in the bathing solution was increased from 6 to 30 mM. When the bathing solution contained 145 mM-potassium, and the patch pipette, 95 mM, a depolarization to -10 mV from a holding potential of -60 mV evoked an inward current. 6. Outward currents were examined by using voltage pulses which depolarized the cell to -20 mV, or more positive values, from a holding potential of -80 mV and by pulses which depolarized the cell to 0 mV, or to positive values, from a holding potential of -30 mV. A Boltzmann fit of typical activation data gave a half-maximal activation voltage of 17 mV and a 'slope' of 14 mV. 7. The time course of the rising phase of the current was described by a function of the form A(1-exp[-(t-delta t)/tau]), where delta t varied between 1 and 4 ms and tau varied between 4 and 27 ms, decreasing with increasing depolarization. There was no evidence for a fast transient component. 8. The amplitude of outward currents was reduced by extracellular calcium ions, cobalt ions, tetraethylammonium and 4-aminopyridine.(ABSTRACT TRUNCATED AT 400 WORDS)