Biomedical subjects

Masayasu Hiraoka

Publications and source records attributed to Masayasu Hiraoka.

At least 19 recordsLinked to original sources

PubMed2006

Hypotonic stress enhances slope conductivity of ATP-sensitive K+ channels activated pharmacologically.

The aim of this paper was to find out the effects of hypotonic stress on the slope conductivity of ATP-sensitive K(+) channels. Using patch clamp technique, rilmakalim and pinacidil as the activators and glibenclamide as an inhibitor of mentioned channels, we have demonstrated that short hypotonic challenge doubled slope conductivity of exploring channels by augmentation of their open probability.

Adenosine Triphosphate↗

PubMed2006

Human cardiac ryanodine receptor mutations in ion channel disorders in Japan.

Catecholaminergic polymorphic ventricular tachycardia (CPVT) is characterized by adrenergic induced bidirectional or polymorphic ventricular tachycardias. Some of CPVT families were reported to be associated with cardiac ryanodine receptor gene (RyR2) mutations. However, association between RyR2 and other arrhythmogenic disorders is not clarified. In this study, we analyzed 83 Japanese patients including patients with long-QT syndrome, Brugada syndrome, idiopathic ventricular fibrillation, arrhythmogenic right ventricular cardiomyopathy and CPVT. Genetic screening of RyR2 revealed 3 distinct mutations among 4 families with CPVT (75% of incidence). However, no mutation was found in other groups. This is the first report to demonstrate prevalence of RyR2 mutations in various arrhythmogenic disorders in Japan. RyR2 mutations were detected frequently in CPVT but not in other diseases.

Arrhythmias, Cardiac↗

PubMed2006

Effects of low-dose quinidine on ventricular tachyarrhythmias in patients with Brugada syndrome: low-dose quinidine therapy as an adjunctive treatment.

Quinidine is suggested as an effective agent to suppress ventricular fibrillation (VF) in the Brugada syndrome by inhibiting transient outward K(+) current (Ito) leading to the reduction and abbreviation of the disparity of repolarization in the right ventricular outflow region and ST segment elevation in the right precordial leads of electrocardiogram. We sought to assess the efficacy of low-dose (300-600 mg) quinidine sulfate on the prevention of ventricular fibrillation induction by programmed electrical stimulation (PES) and spontaneous ventricular fibrillation episodes during the subsequent follow-up period. Electrophysiologic study was performed in 14 patients with the Brugada syndrome (14 men, mean age 50 +/- 11 years, range 32-75) before and during the treatment with low-dose quinidine and evaluated the efficacy of the drug therapy. Ventricular fibrillation was induced in all the patients by programmed electrical stimulation at baseline. After oral quinidine administration (300 mg or 600 mg/d), programmed electrical stimulation was repeated. Ventricular fibrillation induction was prevented in 6 of 14 patients (44%). Serum quinidine concentration was higher in the patients with suppressed VF induction than those without (1.88 +/- 0.44 versus 1.31 +/- 0.43 microg/ml, respectively). After programmed electrical stimulation, 9 of 14 patients (64%), in whom four had implantable cardioverter defibrillator implantation, continued to receive quinidine. During a mean follow-up period of 31 months on quinidine, no side effects except one with diarrhea were observed (12.5%). There were no ventricular fibrillation recurrences in 3 of the 9 patients, who had frequent implantable cardioverter defibrillator discharges due to ventricular fibrillation attacks before treatment with quinidine. Low-dose quinidine has a potential as an adjunctive therapy for patients of the Brugada syndrome with frequent implantable cardioverter defibrillator discharges.

Adult↗

PubMed2006

Metabolic pathways for ion homeostasis and persistent Na(+) current.

ATP supply in heart cells is preserved by a number of different mechanisms to maintain a constant level of ATP concentration. ADP, phosphocreatine, inorganic phosphate, and cAMP-activated kinases are effectively involved in ATP supply in abnormal conditions such as ischemia. Intracellular Na(+) and Ca(2+) concentrations in the heart cells are maintained at low levels through the operation of ion transport across the plasma membrane, such as Na(+)-K(+) pumps, as well as Na(+)-Ca(2+), and Na(+)-H(+) exchangers. The activity of the latter two exchanger mechanisms depends on their expression levels and the concentration gradients of Na(+) and Ca(2+) across the membrane. These exchangers may interact, and both are strongly regulated by intracellular Na(+), which is maintained by the Na(+)-K(+) pump, utilizing ATP as an energy source. The mitochondria and the sarcoplasmic reticulum are the organelles responsible for intracellular Ca(2+) stores and release sites for maintaining very low cytoplasmic concentration of Ca(2+). Ischemia disrupts the delicate interactions of these transport mechanisms. This may cause intracellular Na(+) and Ca(2+) accumulation, which can cause decreased contractility and electrical instability. Further, a persistent (noninactivating) Na(+) current may be enhanced during ischemia, and this can contribute to action potential prolongation, and the development of early afterdepolarizations. The Na(+) influx via the persistent Na(+) current may induce further Na(+) and Ca(2+) accumulation in the cells.

Adenosine Triphosphate↗

PubMed2005

A novel mutation in FKBP12.6 binding region of the human cardiac ryanodine receptor gene (R2401H) in a Japanese patient with catecholaminergic polymorphic ventricular tachycardia.

Catecholaminergic polymorphic ventricular tachycardia (CPVT) is an autosomal dominant inherited disorder characterized by adrenergic induced polymorphic ventricular tachycardias and associated with sudden cardiac death. The human cardiac ryanodine receptor gene (RyR2) was linked to CPVT. A 20-year-old male was referred to our hospital because of recurrent syncope after physical and emotional stress. Routine cardiac examinations including catheterization revealed no structural abnormality. Exercise on treadmill induced premature ventricular contraction in bigeminy and bidirectional ventricular tachycardia was induced during isoproterenol infusion. Beta-blocking drug was effective in suppressing the arrhythmias. We performed genetic screening by PCR-SSCP method followed by DNA sequencing, and a novel missense mutation R2401H in RyR2 located in FKBP12.6 binding region was identified. This mutation was not detected in 190 healthy controls. Since FKBP12.6 plays a critical role in Ca channel gating, the R2401H mutation can be expected to alter Ca-induced Ca release and E-C coupling resulting in CPVT. This is the first report of RyR2 mutation in CPVT patient from Asia including Japan.

Adrenergic beta-Agonists↗

PubMed2005

Spontaneous T wave alternans in a patient with Brugada syndrome--responses to intravenous administration of class I antiarrhythmic drug, glucose tolerance test, and atrial pacing.

Spontaneous T wave alternans in Brugada syndrome. A 43-year-old man with an episode of syncope showed ECG patterns of coved-type ST elevation in leads V1-V3 and right bundle branch block pattern. The patient had spontaneous T wave alternans at baseline, and T wave alternans diminished with distinct development of ST elevation after administration of Na+ channel blocker, and during oral glucose load and atrial pacing. Na+ channel mutation may contribute to the genesis of his ECG changes.

Adult↗

PubMed2005

Modulation of ICa-L by alpha1-adrenergic stimulation in rat ventricular myocytes.

We found when L-type calcium current (ICa-L) was recorded with the perforated patch-clamp method in rat ventricular myocytes that bath application of phenylephrine (with propranolol) evoked a biphasic response characterized by an initial transient suppression followed by a sustained potentiation. The transient suppression occurred 30-60 s after phenylephrine perfusion and reached peak inhibition at approximately 2 min. The biphasic modulation of ICa-L was also elicited by methoxamine, and the effects of phenylephrine were blocked by prazosin, indicating that the responses were mediated through alpha1-adrenoceptors. Pretreatment of cells with H7 (100 micromol/L), a broad-spectrum protein kinase inhibitor that inhibits both protein kinase C and A, eliminated potentiation but did not affect transient suppression. The transient suppression occurred concurrently with the acceleration of the fast component of ICa-L inactivation. Depletion of intracellular Ca2+ stores by ryanodine plus caffeine or thapsigargin eliminated the transient suppression. When ICa-L was recorded with whole-cell patch-clamp and with 0.05 mmol/L EGTA in the pipette solution to allow intracellular Ca2+ to fluctuate, phenylephrine evoked a transient suppression as in the perforated patch recordings. Heparin, a specific blocker of IP3 (inositol 1,4,5-trisphosphate) receptors, eliminated the phenylephrine-induced transient suppression of ICa-L when added to the pipette solution. Intensive chelation of intracellular Ca2+ by 5 mmol/L BAPTA (1,2-bis(2-aminophenoxy)ethane-N,N,N',N'-tetraacetic acid) in the pipette solution also eliminated the phenylephrine-induced transient suppression of ICa-L. We conclude that transient increase in the concentration of intracellular calcium ([Ca2+]i) caused by Ca2+ release from intracellular stores underlies the transient suppression of ICa-L, whereas the potentiation of ICa-L is a result of activation of protein kinases.

1-(5-Isoquinolinesulfonyl)-2-Methylpiperazine↗

PubMed2005

Reversible left ventricular dysfunction (takotsubo cardiomyopathy) with deep negative T waves due to possible cardiac sympathetic denervation.

A case of characteristic left ventricular contraction disorder representing takotsubo cardiomyopathy after exercise is reported. A 58-year-old Japanese woman with no history of cardiac disease was admitted for chest oppression after exercise. A diagnosis of myocardial infarction was suspected because of her symptom, and the electrocardiographic and echocardiographic findings. Coronary angiography revealed normal arteries, while left ventriculography demonstrated asynergy with apical akinesis and basal hyperkinesis. Deep negative T waves were observed on the electrocardiogram on the second hospital day and sustained for over a month. Repeated measurements of heart rate viability and 123I-metaiodobenzylguanidine scintigraphy showed transient dysfunction of the cardiac autonomic nervous system. From these findings, it was concluded that cardiac autonomic denervation plays a pivotal role in the etiology of takotsubo cardiomyopathy.

Cardiomyopathies↗

PubMed2004

Truncated KCNQ1 mutant, A178fs/105, forms hetero-multimer channel with wild-type causing a dominant-negative suppression due to trafficking defect.

We identified a novel mutation Ala178fs/105 missing S3-S6 and C-terminus portions of KCNQ1 channel. Ala178fs/105-KCNQ1 expressed in COS-7 cells demonstrated no current expression. Co-expression with wild-type (WT) revealed a dominant-negative effect, which suggests the formation of hetero-multimer by mutant and WT. Confocal laser microscopy displayed intracellular retention of Ala178fs/105-KCNQ1 protein. Co-expression of the mutant and WT also increased intracellular retention of channel protein compared to WT alone. Our findings suggest a novel mechanism for LQT1 that the truncated S1-S2 KCNQ1 mutant forms hetero-multimer and cause a dominant-negative effect due to trafficking defect.

Adolescent↗

PubMed2004

Functional characterization of a trafficking-defective HCN4 mutation, D553N, associated with cardiac arrhythmia.

Hyperpolarization-activated cyclic nucleotide-gated channel 4 gene HCN4 is a pacemaker channel that plays a key role in automaticity of sinus node in the heart, and an HCN4 mutation was reported in a patient with sinus node dysfunction. Expression of HCN4 in the heart is, however, not confined to the sinus node cells but is found in other tissues, including cells of the conduction system. On the other hand, mutations in another cardiac ion channel gene, SCN5A, also cause sinus node dysfunction as well as other cardiac arrhythmias, including long QT syndrome, Brugada syndrome, idiopathic ventricular fibrillation, and progressive cardiac conduction disturbance. These observations imply that HCN4 abnormalities may be involved in the pathogenesis of various arrhythmias, similar to the SCN5A mutations. In this study, we analyzed patients suffering from sinus node dysfunction, progressive cardiac conduction disease, and idiopathic ventricular fibrillation for mutations in HCN4. A missense mutation, D553N, was found in a patient with sinus node dysfunction who showed recurrent syncope, QT prolongation in electrocardiogram, and polymorphic ventricular tachycardia, torsade de pointes. In vitro functional study of the D553N mutation showed a reduced membranous expression associated with decreased If currents because of a trafficking defect of the HCN4 channel in a dominant-negative manner. These data suggest that the loss of function of HCN4 is associated with sinus nodal dysfunction and that a consequence of pacemaker channel abnormality might underlie clinical features of QT prolongation and polymorphic ventricular tachycardia developed under certain conditions.

Amino Acid Sequence↗

PubMed2004

Block of HERG current expressed in HEK293 cells by the Na+-channel blocker cibenzoline.

A Na(+)-channel blocker, cibenzoline, blocks the delayed rectifier potassium current ( I(k)), but its detailed action on the rapidly activating component ( I(kr)) of I(k) encoded by the human ether-a-go-go-related gene ( HERG) has not been clarified. We examined the effects of cibenzoline on stably expressed HERG current in HEK293 cells recorded by the patch-clamp technique of whole-cell configuration. Cibenzoline blocked HERG current expressed in HEK293 cells with IC(50) = 3.7 +/- 0.963 micro M and Hill coefficient = 0.74 +/- 0.12. Voltage-depended activation was shifted in a negative direction by cibenzoline. No block or minor block was induced at test depolarization of -40 to -30 mV, and the block increased with depolarization reaching a plateau at 0 mV without a further increase at positive voltages. Voltage-dependent activation of HERG currents became faster at negative test voltages but there were no changes at positive voltages after cibenzoline. No frequency-dependent block of HERG tail current by cibenzoline after equilibration was noted between 1.33 and 0.2 Hz. Steady-state inactivation of the HERG current was shifted in a negative direction by approximately 8 mV but the time constants of fast inactivation were little affected by cibenzoline. Cibenzoline blocks the I(kr)-like current reconstituted by HERG clone transfection with an IC(50) value comparable to therapeutic concentrations. Cibenzoline has a preferential affinity, at least, to the open state of the HERG channel with a rapid access to the binding site.

Action Potentials↗

PubMed2004

Functional expression of Ca2+ signaling pathways in mouse embryonic stem cells.

Mouse embryonic stem (mES) cells have the potential to differentiate into all types of cells, but the physiological properties of undifferentiated mES cells, including Ca2+ signaling systems, are not fully understood. In this study, we investigated Ca2+ signaling pathways in mES cells by using confocal Ca2+ imaging systems, patch clamp techniques and RT-PCR. The stimulations with ATP and histamine (His) induced a transient increase of intracellular Ca2+ concentration ([Ca2+]i), which were prevented by the pretreatment of 2-amino-ethoxydiphenyl borate (2-APB), a blocker for inositol-1,4,5-triphosphate receptors (InsP3Rs). The application of caffeine (Caff) or ryanodine (Ry) did not change [Ca2+]i. When stores were depleted with Ca2+ -ATPase blocker, thapsigargin (TG), or histamine, the capacitative Ca2+ entry (CCE) was observed. In whole cell patch clamp mode, store-operated Ca2+ currents could be recorded in cells treated with histamine and thapsigargin. On the other hand, voltage-operated Ca2+ channels (VOCCs) could not be elicited. The application of blockers for plasma membrane Ca2+ pump (PMCAs) (carboxeosin or caloxin2A1) induced a large increase of [Ca2+]i. When the Na+/Ca2+ exchangers (NCXs) were blocked by Na+ free solution or KBR7943, [Ca2+]i was also elevated. Using RT-PCR, mRNAs for InsP3Rs type-1, -2, and -3, PMCA-1 and -4, NCX-1, -2, and -3 could be detected. From these results, we conclude that Ca2+ release from ER is mediated by InsP3Rs in mES cells before differentiation and Ca2+ entry through plasma membrane is mainly mediated by the store-operated Ca2+ channels (SOCs). For the Ca2+ extrusion systems, both NCXs and PMCAs play important roles for maintaining the low level of [Ca2+]i.

Animals↗

PubMed2004

Changes in body surface potential distributions induced by isoproterenol and Na channel blockers in patients with the Brugada syndrome.

BACKGROUND: The characteristics of unique ECG findings in the Brugada syndrome have not been well explained. METHODS: To clarify their characteristics and mechanisms, body surface maps (BSM) were recorded from patients with the Brugada syndrome (13 cases; a mean age of 48 years) before and after administration of isoproterenol (ISP) or Na channel blockers (12 cases). RESULTS: ST elevation in V1-V3 was decreased by 0.1 mV or more after ISP infusion in 8 of 11 cases and elevated after Na channel blockers in 8 of 12. In ventricular activation time (VAT) isochronal map, delayed conduction was noted on upper anterior chest in 11 and on anterior left chest in two. Delayed conduction areas were decreased by ISP and expanded by Na channel blockers. QRST isointegral map showed normal findings in baseline with minimal changes after ISP or Na channel blockers. Activation recovery interval (ARI) isochronal map showed prolonged area on upper anterior chest in baseline, being reduced by ISP and expanded by Na channel blockers. ARI dispersion (ARI-d), defined as difference between the maximum and minimum value of ARI, was larger in Brugada patients than that of normal subjects in baseline, and decreased after ISP and increased after Na channel blockers. CONCLUSION: ST elevation in the Brugada syndrome is primarily caused by abnormality in depolarization rather than in repolarization. BSM can provide better information to clarify a mechanism of ECG changes adding its diagnostic value for this unique syndrome.

Adrenergic beta-Agonists↗

PubMed2004

Novel C-terminus frameshift mutation, 1122fs/147, of HERG in LQT2: additional amino acids generated by frameshift cause accelerated inactivation.

OBJECTIVE: The function of the C-terminus region of the human ether-a-go-go related gene (HERG) has not been well characterized except for its involvement in trafficking. To understand further the role of C-terminus region, we performed a functional analysis of a novel frameshift mutation (1122fs/147) identified in a Japanese long QT syndrome 2 (LQT2) patient who had recurrent episodes of syncope. METHODS: Wild type (WT) and mutant HERG plasmids were transfected into human embryonic kidney (HEK-293) cells, and whole-cell current was recorded by the patch-clamp technique. Confocal microscopy was performed to examine the membrane distribution of channel protein using a green fluorescent protein tagged to the N-terminus of HERG. RESULTS: The mutant 1122fs/147 alone could express current, but reduced density by 74% of control. No dominant negative effect was noted with co-expression of WT and 1122fs/147. Activation and deactivation time constants were not changed, while inactivation was accelerated in 1122fs/147 compared to WT, and V(1/2) of steady-state inactivation curve shifted by 11 mV in the negative direction. Current density of 1123stop mutant revealed 49% reduction compared to WT and showed no shift in steady-state inactivation. Confocal microscopy revealed reduced protein expression on the cell surface both in 1122fs/147 and 1123stop mutants compared to WT. CONCLUSION: Frameshift mutation at the C-terminus region with additional 147 amino acids evoked a loss of function of the HERG channel. A negative shift in steady-state inactivation induced by the additional 147 amino acids and trafficking defect contribute to a reduced current amplitude of 1122fs/147.

Adolescent↗

PubMed2004

Resolution of abnormal body surface maps in children with atrial septal defect after intracardiac repair.

INTRODUCTION: The genesis of repolarization abnormalities of ECG waveforms in atrial septal defect (ASD), which typically is characterized by right ventricular (RV) volume overload, has not been explored, particularly in association with postoperative hemodynamic improvement. The aim of this study was to evaluate the effects of reduced RV overload after ASD closure on depolarization and repolarization abnormalities on body surface maps (BSMs). METHODS AND RESULTS: BSMs of 14 children with ASD were recorded preoperatively and at early postoperative (1-6 months) and late postoperative (>9 months) stages. BSMs of 31 age-matched healthy children were studied as normal controls. Before intracardiac repair, QRS isopotential maps of children with ASD showed delayed RV breakthrough and subsequent rightward enlargement of the positive area with a maximum shifting to the right. Delayed conduction of the RV, particularly at the outflow tract area, was noted. The preoperative QRST isointegral maps exhibited the two-maximum pattern reflecting repolarization abnormality. The delayed appearance of breakthrough and delayed RV conduction on the QRS isopotential maps persisted from the preoperative to the late postoperative stage, whereas the two-maximum pattern on the QRST isointegral maps normalized to the one-dipole pattern at an early stage after repair. CONCLUSION: Abnormal repolarization parameters in ASD patients showed rapid improvement postoperatively, despite the persistence of depolarization abnormalities. Therefore, the two-maximum pattern on the QRST isointegral maps indicates a primary T wave change due to hemodynamic RV volume overload.

Adolescent↗

PubMed2004

Unexpected mexiletine responses of a mutant cardiac Na+ channel implicate the selectivity filter as a structural determinant of antiarrhythmic drug access.

Gating properties of Na(+) channels are the critical determinants for the state-dependent block by class I antiarrhythmic drugs; however, recent site-directed mutagenesis studies have shown that the Na(+) channel selectivity filter region controls drug access to and dissociation from the binding site. To validate these observations, we have exploited a naturally occurring cardiac Na(+) channel mutation, S1710L, located next to the putative selectivity filter residue of domain 4, and evaluated the pharmacological properties to mexiletine using whole-cell, patch-clamp recordings. Consistent with the large negative shift of steady-state inactivation and the enhanced slow inactivation, the S1710L channel showed greater mexiletine tonic block than wild-type (WT) channel. In contradiction, S1710L showed attenuated use-dependent block by mexiletine and accelerated recovery from block, suggesting that the drug escape though the external access path is facilitated. Extracellularly applied QX-314, a membrane-impermeant derivative of lidocaine, elicited significantly enhanced tonic block in S1710L similar to mexiletine. However, recovery from internally applied QX-314 was accelerated by 4.4-fold in S1710L compared with WT. These results suggest that the drug access to and dissociation from the binding site through the hydrophilic path are substantially altered. Moreover, K(+) permeability was 1.9-fold increased in S1710L, verifying that the mutated residue is located in the ion-conducting pore. We propose that the Na(+) channel selectivity filter region is a structural determinant for the antiarrhythmic drug sensitivity in addition to gating properties of the indigenous Na(+) channels that govern the state-dependent drug block.

Amino Acid Substitution↗

PubMed2004

The effects of K+ channels modulators terikalant and glibenclamide on membrane potential changes induced by hypotonic challenge of guinea pig ventricular myocytes.

Contribution of inward rectifier K(+) currents (I(K1)) and ATP-sensitive K(+) currents (I(KATP)) to membrane potential changes of ventricular myocytes appearing during hypotonic challenge is unclear. We used here the whole cell patch clamp technique, voltage and current clamp modes, to record membrane potentials and ionic currents in isolated guinea pig ventricular myocytes under isotonic or hypotonic perfusion. The difference in osmolarity between iso- and hypotonic solutions was about 100 mOsm. Exposure to hypotonic solution for 60 s induced initial prolongation of action potential duration at 90% of repolarization (APD(90)) (from 176 +/- 10 to 189 +/- 11 ms, P<0.05, n = 13). Further perfusion for the next 300 s shorthened APD(90) to 135 +/- 9 ms (P<0.01, in comparison with control values, n = 13) and depolarized resting potential from -79.2 +/- 1.5 to -75.0 +/- 0.9 mV, (P<0.05, n = 13). Neither pretreatment with a blocker of I(K1) channels, terikalant at 10 microM, nor with a blocker of I(KATP) channels, glibenclamide at 1 microM, prevented the above-mentioned changes in membrane potential induced by hypotonic challenge when a pipette solution containing 5 mM ATP was used. Also, glibenclamide and terikalant did not affect the hypotonic-sensitive current, obtained by ramp or voltage-step protocols, respectively. Additionally, the current-voltage relationship (I-V curve) of the whole cell hypotonic-sensitive current shifted from an isotonic I-V curve in a parallel way. Our results indicate that I(K1) and I(KATP) do not participate in membrane potential changes induced by hypotonic solution at least in the guinea pig ventricular myocytes with sufficient intracellular ATP.

Animals↗

PubMed2004

Hypotonic stress increases efficacy of rilmakalim, but not pinacidil, to activate ATP-sensitive K(+) current in guinea pig ventricular myocytes.

The aim of this study was to investigate the influence of hypotonic challenge on the effects of potassium channel openers (PCO), rilmakalim and pinacidil, on activation of the ATP-sensitive K(+) current. The whole cell configuration of the patch-clamp technique was applied to guinea pig ventricular myocytes exposed to isotonic and hypotonic solutions. Difference in osmolarity was about 100 mOsm due to different mannitol concentrations. Rilmakalim, a second generation PCO [(3S,4R)-3-hydroxy-2,2-dimethyl-4-(2-oxo-l-pyrrolidinyl)-6-phenylsulfonylchromanhemihydrate], activated time-independent K(+) current in the isotonic solution with pD2 (-log EC(50)) = 6.42 +/- 0.12 and E(max) = 19.74 +/- 2,16 pA/pF, n = 7, at 0 mV. The effects of the cyanoguanidine compound pinacidil were similar to those of rilmakalim, but the action appeared slower and with about 600-fold less potency than the former. Efficacy of rilmakalim, but not pinacidil, was enhanced in hypotonic solution, with E(max) = 30.87 +/- 5.40 pA/pF (P<0.05, n = 7), and the current was completely inhibited by glibenclamide. Additionally, rilmakalim concentration-effects correlation coefficient (R) decreased from 0.96 to 0.86 and Hill's coefficient increased from 1.21 to 1.45. Pretreatment with phalloidin (20 microM), a cytoskeleton stabilizer, prevented an intensification of the effects of rilmakalim in hypotonic solution and returned R and Hill's coefficients to the control values. We conclude that osmotic stress increases efficacy of rilmakalim to activate K(ATP) channels in guinea pig ventricular myocytes due to the specific interaction with actin filaments.

ATP-Binding Cassette Transporters↗