SEARCH · PubMed Health

Results for “Potassium Channel Blockers”

Explore indexed PubMed citations for clinical trials, systematic reviews and public health research. Read source abstracts and follow each citation to its original PubMed record.

Quote a phrase for an exact phrase match. Source license links do not imply unrestricted reuse.

At least 289 records · Page 16Linked to original sources

PubMed1996

Effect of capsaicin and analogues on potassium and calcium currents and vanilloid receptors in Xenopus embryo spinal neurones.

1. The potassium current in embryo spinal neurones of Xenopus consists of at least two kinetically distinct components with overlapping voltage-dependencies of activation. We investigated whether capsaicin might specifically block these components in acutely dissociated neurones from stage 37/38 embryos by use of standard patch clamp techniques. 2. Capsaicin caused a time-dependent block of both the slow and fast components of the potassium current. The concentration-dependence was described by the Hill equation with a KD of 21 microM and a coefficient of 1.5 (n = 9-11 at each concentration). Differences between the observed and fitted values were not significant at the 5% level (chi(2) = 2.80, 6 degrees of freedom). 3. Capsaicin did not affect the time course or voltage-sensitivity of activation, but the steady-state block was voltage-dependent. The block could be relieved by hyperpolarization, and the rate of the removal of block was voltage- and time-dependent. The time constant for the blocking reaction was also voltage-dependent for voltage steps below +30 mV, but above this level it was voltage-independent. These results suggest that capsaicin blocks potassium channels by an open channel mechanism. 4. Other derivatives of vanillin, such as capsazepine, resiniferatoxin, and piperine also blocked potassium channels. Capsazepine and resiniferatoxin caused a greater block than similar concentrations of capsaicin, and in the case of capsazepine, the block was also clearly time-dependent. 5. Capsaicin and capsazepine also blocked calcium currents in a time-dependent manner. Fitting the Hill equation to the averaged data gave a KD of 43.5 microM, and a coefficient of 1.35 (n = 11 at each concentration). The fitted values were not significantly different from the observed means at the 5% level (chi(2) = 12.1, 6 degrees of freedom). 6. Six out of 29 Rohon-Beard sensory neurones responded to capsaicin with an inward current that appeared to be similar to the capsaicin activation of mammalian C sensory neurones. This response saturated at 10 microM capsaicin.

Animals↗

PubMed1994

On the mechanism of 4-aminopyridine action on the cloned mouse brain potassium channel mKv1.1.

1. This study used the whole-cell patch clamp technique to investigate the mechanism of action of the K+ channel blocker 4-aminopyridine (4-AP) on the cloned K+ channel mouse Kv1.1 (mKv1.1) expressed in Chinese hamster ovary cells. 2. Cells transfected with mKv1.1 expressed a non-inactivating, delayed rectifier-type K+ current. 4-AP induced a dose-, voltage- and use-dependent block of mKv1.1. 3. 4-AP blockade of mKv1.1 was similar whether 4-AP was administered extracellularly (IC50 = 147 microM) or intracellularly (IC50 = 117 microM). 4. Inclusion of the first twenty amino acids of the N-terminus sequence of the Shaker B K+ channel ('inactivation peptide') in the patch electrode transformed mKv1.1 into a rapidly inactivating current. The time constant of decay for the modified current was dependent on the concentration of inactivation peptide, and under these conditions extracellular 4-AP had a reduced potency (IC50 values of 471 and 537 microM for 0.5 and 2 mg ml-1 inactivation peptide, respectively). 5. A permanently charged analogue of 4-AP, 4-aminopyridine methiodide (4-APMI), was found to block mKv1.1 when applied inside the cell, but was without effect when administered externally. 6. Decreasing the intracellular pH (pHi) to 6.4 caused an increase in 4-AP potency (IC50 = 76 microM), whereas at pHi 9.0, the 4-AP potency fell (IC50 = 295 microM). Conversely, increasing extracellular pH (pHo) to 9.0 caused an increase in 4-AP potency (IC50 = 93 microM), whereas at pHo 6.4, 4-AP potency decreased (IC50 = 398 microM). 7. Taken together, these findings support the hypotheses that the uncharged form of 4-AP crosses the membrane, and that it is predominantly the cationic form which acts on mKv1.1 channels intracellularly, possibly at or near to the binding site for the inactivation peptide.

4-Aminopyridine↗

PubMed2000

A subset of periglomerular neurons in the rat accessory olfactory bulb may be excited by GABA through a Na(+)-dependent mechanism.

The periglomerular (PG) cells of the accessory olfactory bulb (AOB) are GABAergic interneurons which receive input from the vomeronasal sensory neurons and form dendrodendritic synapses with each other and with mitral cells. Their electrophysiological properties have not been investigated. We have developed a novel method of isolating PG cells from the AOB, and used the whole-cell patch and gramicidin-perforated patch clamp techniques to measure their basic electrophysiological characteristics and their response to GABA. PG cells were found to be excitable neurons with voltage-gated Na(+) and K(+) currents, though it was very difficult to get PG cells to fire an action potential. The voltage-gated Na(+) currents of PG cells activate at more positive potentials than those of typical CNS neurons. PG cells respond to GABA with currents in which GABA(A) receptors play a significant role. A subset ( approximately 40%) of PG cells respond to GABA with currents which have unusually high reversal potentials, indicating that GABA may be excitatory to these neurons. This phenomenon cannot be explained entirely by elevation of intracellular chloride concentrations, and is dependent on the presence of extracellular sodium.

Action Potentials↗

PubMed2002

Inhibition of cardiac Na+ current by primaquine.

The electrophysiological effects of the anti-malarial drug primaquine on cardiac Na(+) channels were examined in isolated rat ventricular muscle and myocytes. In isolated ventricular muscle, primaquine produced a dose-dependent and reversible depression of dV/dt during the upstroke of the action potential. In ventricular myocytes, primaquine blocked I(Na)(+) in a dose-dependent manner, with a K(d) of 8.2 microM. Primaquine (i) increased the time to peak current, (ii) depressed the slow time constant of I(Na)(+) inactivation, and (iii) slowed the fast component for recovery of I(Na)(+) from inactivation. Primaquine had no effect on: (i) the shape of the I - V curve, (ii) the reversal potential for Na(+), (iii) the steady-state inactivation and g(Na)(+) curves, (iv) the fast time constant of inactivation of I(Na)(+), and (v) the slow component of recovery from inactivation. Block of I(Na)(+) by primaquine was use-dependent. Data obtained using a post-rest stimulation protocol suggested that there was no closed channel block of Na(+) channels by primaquine. These results suggest that primaquine blocks cardiac Na(+) channels by binding to open channels and unbinding either when channels move between inactivated states or from an inactivated state to a closed state. Cardiotoxicity observed in patients undergoing malaria therapy with aminoquinolines may therefore be due to block of Na(+) channels, with subsequent disturbances of impulse conductance and contractility.

Action Potentials↗

PubMed1996

Sodium and potassium currents of type I spiral ganglion cells from rat.

Ion channel activity of acutely dissociated type I spiral ganglion cells isolated from rats was investigated using the whole-cell variation of the patch clamp technique. Tetrodotoxin-sensitive sodium current and tetraethylammonium-sensitive potassium current were recorded. With a holding potential of -80 mV, peak sodium currents were generated by depolarizations to membrane potentials more positive than -50 mV. Potassium currents were elicited at membrane potentials more positive than -40 mV. Some cells which did not exhibit sodium current exhibited delayed rectifier potassium currents. Steady-state sodium channel inactivation curve yielded a slope of 12 mV and a half-inactivated potential of -83 mV. Recovery from inactivation of the sodium channel proceeded with a fast and slow time course; recovery began as early as 0.8 ms and proceeded with a time constant of 7.5 ms. It is concluded that type I spiral ganglion cells are endowed with sodium and potassium channels which are responsible for generation and propagation of auditory nerve action potentials.

Action Potentials↗

PubMed2000

Characterization of mibefradil block of the human heart delayed rectifier hKv1.5.

The goal of this study was to analyze the effects of mibefradil on a human cardiac K(+) channel (hKv1.5) stably expressed in Chinese hamster ovary cells using the whole-cell configuration of the patch-clamp technique. Mibefradil inhibited in a concentration-dependent manner the hKv1.5 current with a K(D) value of 0.78 +/- 0.05 microM and a Hill coefficient of 0.97 +/- 0.06. Block induced by mibefradil was voltage dependent, consistent with a value of electrical distance of 0.13. The apparent association (k) and dissociation (l) rate constants measured at +50 mV were found to be 7.3 +/- 0.5 x 10(6) M(-1).s(-1) and 4.3 +/- 0.1 s(-1), respectively. Block increased rapidly between -20 and +10 mV, coincident with channel opening and suggested an open channel block mechanism, which was confirmed by a slower deactivation time course resulting in a "crossover" phenomenon when tail currents recorded under control conditions and in the presence of mibefradil were superimposed. Shifts toward negative potentials of the maximum conductance and the activation curve were observed, confirming the voltage dependence of block. Mibefradil induced a significant use-dependent block when trains of depolarization at frequencies between 0.02 and 2 Hz were applied. In the presence of mibefradil, recovery of inactivation was faster than under control conditions, suggesting that mibefradil might compete with the inactivation gate of hKv1.5. These results indicate that mibefradil blocks hKv1.5 channels in a concentration-, voltage-, time- and use-dependent manner and the concentrations needed to observe these effects are in the therapeutic range.

Animals↗

PubMed1998

Role of potassium channels in the nitrergic nerve stimulation-induced vasodilatation in the guinea-pig isolated basilar artery.

1. We studied the effects of various K+ channel blockers on the vasodilator responses of guinea-pig isolated basilar arteries to nitrergic nerve stimulation, the nitric oxide (NO) donor sodium nitroprusside (SNP), and the membrane permeable guanosine-3',5'-cyclic monophosphate (cyclic GMP) analogue 8-bromo-cyclic GMP (8-Br-cyclic GMP). 2. In endothelium-denuded preparations which were contracted with prostaglandin F2alpha (1 microM), electrical field stimulation (EFS, 10 Hz for 30 s) produced a vasodilatation which was totally blocked by the nitric oxide synthase (NOS) inhibitor N(G)-nitro-L-arginine methyl ester L-NAME; 100 microM) (n=3) and by the selective NO-sensitive guanylate cyclase inhibitor 1H-[1,2,4]oxadiazolo[4,3,-a]quinoxalin-1-one (ODQ; 1 microM) (n=4). The vasodilator response to SNP (100 nM) was not reduced by L-NAME but was abolished by ODQ (1 microM) (n=4). 3. EFS-elicited vasodilatation was partly but significantly reduced by the non-selective K+ channel blockers tetraethylammonium (TEA, 1 and 3 mM) and 4-aminopyridine (4-AP, 3 mM), and by the large-conductance calcium-activated K+ channel (K(Ca) channel) blockers charybdotoxin (ChTX, 150 nM) and iberiotoxin (IbTX, 30 and 100 nM). In contrast, the ATP-sensitive K+ channel (K(ATP) channel) blocker glibenclamide (1-10 microM) and the small-conductance K(Ca) channel blocker apamin (100-500 nM) did not affect EFS-induced vasodilatation. 4. The vasodilator response elicited by SNP (10-100 nM) was significantly reduced by TEA (3 mM) and ChTX (150 nM) but not by apamin (500 nM) or glibenclamide (1 microM). The vasodilatation elicited by 8-Br-cyclic GMP (100 microM) was also reduced by TEA (3 mM) and ChTX (150 nM). 5. The results indicate that the vasodilatations induced by nitrergic nerve stimulation and the NO donor SNP in endothelium-denuded guinea-pig basilar artery depend on the formation of intracellular cyclic GMP. The increased cyclic GMP level activates large-conductance K(Ca) channels which partly mediate the vasodilator response. Neither K(ATP) channels nor apamin-sensitive small-conductance K(Ca) channels are involved in nitrergic transmitter-mediated vasodilatation.

Animals↗

PubMed2006

Potassium channels-mediated vasorelaxation of rat aorta induced by resveratrol.

Resveratrol, a phenolic substance present in grapes and a variety of medical plants, has been reported to induce vasorelaxation, however the mechanisms are uncertain. In this paper we investigate the possible participation of K(+) channels in the endothelium-independent vasodilatation of rat aorta induced by resveratrol. Resveratrol induced concentration-dependent relaxation of rings with endothelium and without endothelium. We used different potassium channel inhibitors to determine whether the K(+) channels mediated endothelium-independent relaxation of rat aorta induced by resveratrol. Highly selective blocker of ATP-sensitive K(+) channels, glibenclamide, as well as non-selective blockers of K(+) channels, tetraethylammonium, did not block resveratrol-induced relaxation of rat aortic rings. Charybdotoxin, a blocker of calcium-sensitive K(+) channels did not affect the resveratrol-induced relaxation. 4-Aminopiridine, non-selective blocker of voltage-gated K(+) (Kv) channels, and margatoxin that inhibits Kv1 channels abolished relaxation of rat aortic rings induced by resveratrol. In conclusion, we have shown that resveratrol potently relaxed rat aortic rings with denuded endothelium. It seems that 4-aminopiridine and margatoxin-sensitive K(+) channels located in the smooth muscle of rat aorta mediated this relaxation.

4-Aminopyridine↗

PubMed1996

Oxygen-induced constriction of rabbit ductus arteriosus occurs via inhibition of a 4-aminopyridine-, voltage-sensitive potassium channel.

The ductus arteriosus is a vital fetal structure allowing blood ejected from the right ventricle to bypass the pulmonary circulation in utero. Closure of the ductus arteriosus at birth, essential for postnatal adaptation, is initiated by an increase in oxygen (O2) tension. We recently demonstrated the presence of O2-sensitive potassium channels in the fetal and adult pulmonary circulation which regulate vascular tone in response to changes in O2 tension. In this study, we assessed the cellular mechanisms underlying O2-induced constriction of the ductus arteriosus in late-gestation fetal rabbits. We report that O2 reversibly inhibits a 58-pS voltage- and 4-aminopyridine-sensitive potassium channel, causing membrane depolarization, an increase in intracellular calcium through L-type voltage-gated calcium channels, and constriction of the ductus arteriosus. We conclude that the effector mechanism for O2 sensing in the ductus arteriosus involves the coordinated action of delayed rectifier potassium channels and voltage-gated calcium channels.

4-Aminopyridine↗

PubMed2005

Systematic administration of iptakalim, an ATP-sensitive potassium channel opener, prevents rotenone-induced motor and neurochemical alterations in rats.

Our previous studies revealed that iptakalim, a novel ATP-sensitive potassium channel opener, has a significant neuroprotective function against ischemia in vivo or rotenone-induced neurotoxicity in vitro. To investigate the potential pharmaceutical benefit of ATP-sensitive potassium channel openers on neurodegenerative diseases, we studied the effects of iptakalim and diazoxide, a selective mitochondrial ATP-sensitive potassium channel opener, on the rotenone-induced nigrostriatal degeneration in rats. Iptakalim (1.5 mg/kg/day, orally) or diazoxide (1.5 mg/kg/day, orally) alone was administered to rats for 3 days, and then for 4 weeks was used daily with an injection of rotenone (2.5 mg/kg/day, subcutaneously) 1 hr later each time. The results showed that rotenone-infused rats exhibited parkinsonian symptoms and had dopamine depletion in the striatum and substantia nigra. Pretreatment with iptakalim or diazoxide prevented rotenone-induced catalepsy and the reduction of striatum dopamine contents. Moreover, iptakalim and diazoxide reduced the enzymatic activities and mRNA levels of inducible nitric oxide synthase elicited by chronic administration of rotenone. These neuroprotective effects of iptakalim and diazoxide were abolished by 5-hydroxydecanoate, a selective mitochondrial ATP-sensitive potassium channel blocker. In conclusion, our data suggested that mitochondrial ATP-sensitive potassium channels might play a key role in preventing both parkinsonian symptoms and neurochemistry alterations induced by rotenone in rats. The selective activation of mitochondrial ATP-sensitive potassium channels may provide a new therapeutic strategy for prevention and treatment of neurodegenerative disorders such as Parkinson's disease.

ATP-Binding Cassette Transporters↗

PubMed2004

Sodium-activated potassium conductance participates in the depolarizing afterpotential following a single action potential in rat hippocampal CA1 pyramidal cells.

The depolarizing afterpotential (DAP) following an action potential increases the excitability of a neuron. Mechanisms related to the DAP following an antidromic or current-induced spike were studied in CA1 pyramidal cells by whole-cell recordings in hippocampal slices in vitro. In DAP-holding voltage curves, the DAP at 10 ms after the spike peak (DAP10) was extrapolated to reverse at about -50 mV. Increase of extracellular K(+) concentration increased DAP and neuronal bursting. DAP10 reversal potential shifted positively with an increase in [K(+)](o) and with the blockade of K(+) conductance using pipettes filled with Cs(+). Similarly, extracellular tetraethylammonium (TEA; 10 mM), 4-aminopyridine (3-10 mM) increased DAP and shifted the DAP10 reversal potential to a depolarizing direction. Decrease of [Ca(2+)](o) did not alter DAP significantly, suggesting a nonessential role of Ca(2+) in the DAP. Perfusion of tetrodotoxin (TTX; 0.1-1 microM) and replacement of extracellular Na(+) by choline(+) suppressed both spike height and DAP simultaneously. Replacement of extracellular Na(+) by Li(+) increased DAP and spike bursts, and caused a positive shift of the DAP10 reversal potential. It is suggested that Li(+) increased DAP by blocking an Na(+)-activated K(+) current. In summary, multiple K(+) conductances are normally active during the DAP following a single action potential.

4-Aminopyridine↗

PubMed2001

Dendritic coincidence detection of EPSPs and action potentials.

We describe a mechanism for coincidence detection mediated by the interaction between backpropagating action potentials and EPSPs in neocortical pyramidal neurons. At distal dendritic locations, appropriately timed EPSPs or oscillations could increase the amplitude of backpropagating action potentials by three- to fourfold. This amplification was greatest when action potentials occurred at the peak of EPSPs or dendritic oscillations and could lead to somatic burst firing. The increase in amplitude required sodium channel activation but not potassium channel inactivation. The temporal characteristics of this amplification are similar to those required for changes in synaptic strength, suggesting that this mechanism may be involved in the induction of synaptic plasticity.

4-Aminopyridine↗

PubMed1996

Subthreshold membrane resonance in neocortical neurons.

1. Using whole cell recording techniques, we studied subthreshold and suprathreshold voltage responses to oscillatory current inputs in neurons from the sensorimotor cortex of juvenile rats. 2. Based on firing patterns, neurons were classified as regular spiking (RS), intrinsic bursting (IB), and fast spiking (FS). The subthreshold voltage-current relationships of RS and IB neurons were rectifying whereas FS neurons were almost ohmic near rest. 3. Frequency response curves (FRCs) for neurons were determined by analyzing the frequency content of inputs and outputs. The FRCs of most neurons were voltage dependent at frequencies below, but not above, 20 Hz. Approximately 60% of RS and IB neurons had a membrane resonance at their resting potential. Resonant frequencies were between 0.7 and 2.5 Hz (24-26 degrees C) near -70 mV and usually increased with hyperpolarization and decreased with depolarization. The remaining RS and IB neurons and all FS neurons were nonresonant. 4. Resonant neurons near rest had a selective coupling between oscillatory inputs and firing. These neurons selectively fired action potentials when the frequency of the swept-sine-wave (ZAP) current input was near the resonant frequency. However, when these neurons were depolarized to -60 mV, spike firing was associated with many input frequencies rather than selectively near the resonant frequency. 5. We examined three subthreshold currents that could cause low-frequency resonance: IH, a slow, hyperpolarization-activated cation current that was blocked by external Cs+ but not Ba2+; IIR, an instantaneously activating, inwardly rectifying K+ current that was blocked by both Cs+ and Ba2+; and INaP, an quickly activating, inwardly rectifying persistent Na+ current that was blocked by tetrodotoxin (TTX). Voltage-clamp experiments defined the relative steady state activation ranges of these currents. IIR (activates below -80 mV) and INaP (activates above -65 mV) are unlikely to interact with each other because their activation ranges never overlap. However, both currents may interact with IH, which activated variably at potentials between -50 and -90 mV in different neurons. 6. We found that IH produces subthreshold response. Consistent with this, subthreshold resonance was blocked by external Cs+ but not Ba2+ or TTX. Application of Ba2+ enlarged FRCs and resonance at potentials below -80 mV, indicating that IK,ir normally attenuates resonance. Application of TTX greatly diminished resonance at potentials more depolarized than -65 mV, indicating that INaP normally amplifies resonance at these potentials. 7. The ZAP current input may be viewed as a model of oscillatory currents that arise in neocortical neurons during synchronized activity in the brain. We propose that the frequency selectivity endowed on neurons by IH may contribute to their participation in synchronized firing. The voltage dependence of the frequency-selective coupling between oscillatory inputs and spikes may indicate a novel mechanism for controlling the extent of low-frequency synchronized activity in the neocortex.

Animals↗

PubMed1998

Ca2+ and K+ currents regulate accommodation and firing frequency in guinea pig bronchial ganglion neurons.

Intracellular microelectrode recordings were obtained from neurons located in adult guinea pig bronchial parasympathetic ganglia in situ to determine the calcium and potassium currents regulating repetitive action potential activity and firing rates by these neurons. Neurons in these ganglia respond to prolonged suprathreshold depolarizing current steps with either a burst of action potentials at the onset of the stimulus (accommodating or phasic neurons) or repetitive action potentials throughout the stimulus (nonaccommodating or tonic neurons). Instantaneous and adapted firing rates during prolonged threshold and suprathreshold stimuli were lower in tonic than in phasic neurons, indicating a longer interspike interval between repetitive action potentials in tonic neurons. In tonic neurons, blockade of A-type current with 4-aminopyridine increased accommodation; 4-aminopyridine or apamin decreased the interspike interval in tonic neurons. Calcium-free buffer, cadmium ions, or omega-conotoxin GVIA also increased accommodation in tonic neurons but did not affect the interspike interval; nifedipine or verapamil did not affect the tonic firing pattern. Accommodation in phasic neurons could be decreased by a conditioning hyperpolarization step of the resting potential, which could be subsequently blocked by 4-aminopyridine or calcium-free buffer. Accommodation in phasic neurons could also be decreased by apamin or barium ions: the repetitive action potentials observed during these treatments could be reversed by cadmium ions or calcium-free buffer. These results indicate that tonic and phasic neurons in guinea pig bronchial parasympathetic ganglia have similar types of calcium currents, but potassium channels may ultimately regulate the accommodation pattern, the firing rate, and, consequently, the output from these neurons.

4-Aminopyridine↗

PubMed2001

Inhibition of glial Na+ and K+ currents by tamoxifen.

Tamoxifen (tmx) is a non-steroidal triphenylethylene derivative that is predominantly known as a competitive antagonist at the estrogen receptor and is used in the treatment of breast cancer. Recent studies suggest that tamoxifen is also beneficial in the treatment of brain metastases and primary brain tumors. Tmx accumulates in brain and its concentration can be up to 46-fold higher than in serum. Therefore, astrocytes may be exposed to tmx in vivo. We use the whole-cell patch-clamp technique to examine the effects of tmx on voltage-dependent cation currents in rat cortical cultures. Using biophysical and pharmacological methods, we isolate sustained and transient outward potassium currents (I(KS) and I(KT), respectively), inwardly rectifying potassium currents (I(KIR)), and transient inward sodium currents (I(Na)). We show that that TTX-sensitive I(Na) is completely inhibited by 10 microm tmx within 5 min. Similarly, tmx blocks I(KS), but does not inhibit I(KT) or I(KIR) at these concentrations. Tmx effects are irreversible with 10 min wash. Interestingly, the currents sensitive to tmx are important in growth control of glial cells (MacFarlane & Sontheimer, 1997). Therefore, we examine cytotoxic and proliferative effects of tmx. Tmx (10 microm) is not cytotoxic as judged by trypan blue exclusion. However, incubation with tmx significantly reduces cell proliferation as examined by 3[H]-thymidine uptake.

Animals↗

PubMed2003

Calcium-activated transient outward chloride current and phase 1 repolarization of swine ventricular action potential.

OBJECTIVE: It is unknown whether 4-aminopyridine- (4-AP-) sensitive transient outward K(+) current (I(to1)) and/or Ca(2+)-activated transient outward Cl(-) current (I(Ca.Cl) or I(to2)) contribute(s) to phase 1 repolarization of pig ventricular action potential (AP). The purpose of the present study was to determine ionic contribution of the phase 1 repolarization of AP in pig ventricle. METHODS: We used whole-cell patch techniques to record APs and membrane currents, and Western immunoblotting analysis to detect expression of I(to1) protein (Kv4.2 or Kv4.3) in pig ventricular myocytes. RESULTS: A transient outward current (I(to)) was activated upon depolarization voltage steps to between -10 and +60 mV from -50 mV in pig ventricular cells, and the I(to) was resistant to 4-AP application, but sensitive to the inhibition by ryanodine (10 micromol/l) and the Ca(2+) channel blockade, and the Cl(-) channel blocker 4,4'-diisothiocyanostilben-2,2'disulfonic acid (DIDS, 150 micromol/l). The current was diminished by external Cl(-) (Cl(-)(o)) replacement and showed a 'bell-shaped' I-V relationship at room temperature, typical of I(to2). No difference in I(to2) was observed in the regional cells from epicardium, midmyocardium, and endocardium of left ventricle. APs showed significant phase 1 and 'spike and dome' in pig ventricular myocytes. The phase 1 and 'spike and dome' of APs were not affected by 4-AP (3 mmol/l), but abolished by replacing Cl(-)(o) and by application of 100 micromol/l DIDS, suggesting I(to2) contribution. Western immunoblotting analysis showed no evidence for the expression of 4-AP-sensitive I(to1) channel protein (Kv4.2 or Kv4.3) in pig ventricular cells. CONCLUSION: The results indicate that 4-AP-sensitive I(to1) is not expressed, and only Ca(2+)-activated I(to2) is present in pig cardiac cells, which contributes importantly to the phase 1 repolarization of ventricular APs in this species.

4,4'-Diisothiocyanostilbene-2,2'-Disulfonic Acid↗

PubMed2006

Pharmacological Profile of U-37883A, a Channel Blocker of Smooth Muscle-Type ATP-Sensitive K Channels.

U-37883A (PNU-37883A, guanidine; 4-morpholinecarboximidine-N-1-adamantyl-N'-cyclohexyl hydrochloride) was originally developed as a potential diuretic with specific binding in kidney and vascular smooth muscle rather than in brain or pancreatic beta cells. U-37883A inhibits ATP-sensitive K(+) channels (K(ATP) channels) in vascular smooth muscle at submicromolar concentrations whilst even at high concentrations (> or =10 microM) it has no inhibitory effect at pancreatic, cardiac or skeletal K(ATP) channels. Thus, it is generally thought that U-37883A is a selective inhibitor of vascular smooth muscle K(ATP) channels. Approximately one decade ago, K(ATP) channels were cloned and found to consist of at least two subunits: an inwardly-rectifying K(+) channel six family (K(ir)6.x; K(ir)6.1 and K(ir)6.2) which forms the ion conducting pore and a modulatory sulphonylurea receptor (SUR.x; SUR1, SUR2A, and SUR2B) that accounts for several pharmacological properties. It is generally believed that different combinations of K(ir)6.x and SUR.x determine the molecular properties of K(ATP) channels. Thus, K(ir)6.2/SUR1 channel represents the pancreatic beta-cell K(ATP) channel, K(ir)6.2/SUR2A channel is thought to represent the cardiac K(ATP) channel, whereas K(ir)6.1/SUR2B channel is likely to represent the vascular smooth muscle K(ATP) channel. Recent molecular studies have shown that U-37883A selectively suppresses the activity of recombinant K(ATP) channels which contain K(ir)6.1 subunits in the channel pore unit. It was thus thought that U-37883A was a selective pharmacological tool which could be used to investigate the activity of vascular smooth muscle K(ATP) channels. However, due to its multiple pharmacological actions on several ion channels and poor tissue selectivity, U-37883A should not be viewed as a selective blocker of smooth muscle K(ATP) channels.

Adamantane↗

PubMed2002

Hypoxic vasorelaxation inhibition by organ culture correlates with loss of Kv channels but not Ca(2+) channels.

We (Thorne GD, Shimizu S, and Paul RJ. Am J Physiol Cell Physiol 281: C24-C32, 2001) have recently shown that organ culture for 24 h specifically inhibits relaxation to acute hypoxia (95% N(2)-5% CO(2)) in the porcine coronary artery. Here we show similar results in the porcine carotid artery and the rat and mouse aorta. In the coronary artery, part of the inability to relax to hypoxia after organ culture is associated with a concomitant loss in ability to reduce intracellular Ca(2+) concentration ([Ca(2+)](i)) during hypoxia (Thorne GD, Shimizu S, and Paul RJ. Am J Physiol Cell Physiol 281: C24-C32, 2001). To elucidate the mechanisms responsible for the loss of relaxation to hypoxia, we investigated changes in K(+) and Ca(2+) channel activity and gene expression that play key roles in [Ca(2+)](i) regulation in vascular smooth muscle (VSM). Reduced mRNA expression of O(2)-sensitive K(+) channels (Kv1.5 and Kv2.1) was shown by reverse transcriptase-polymerase chain reaction in the rat aorta. In contrast, no change in other expressed voltage-gated K(+) channels (Kv1.2 and Kv1.3) or Ca(2+) channel subtypes was found. Modified K(+) channel expression is supported by functional evidence indicating a reduced response to general K(+) channel activation, by pinacidil, and to specific voltage-dependent K(+) (Kv) channel blockade by 4-aminopyridine. In conclusion, organ culture decreases expression of specific Kv channels. These changes are consistent with altered mechanisms of VSM contractility that may be involved in Ca(2+)-dependent pathways of hypoxia-induced vasodilation.

Animals↗