Ventricular arrhythmogenesis following slowed conduction in heptanol-treated, Langendorff-perfused mouse hearts

Arrhythmogenic effects of slowed action potential conduction produced by the gap junction and sodium-channel inhibitor heptanol (0.1–2 mM) were explored in Langendorff-perfused mouse hearts. Monophasic action potential recordings showed that 2 mM heptanol induced ventricular tachycardia in the absence of triggered activity arising from early or after-depolarizations during regular 8 Hz pacing and programmed electrical stimulation (PES). It also increased activation latencies and ventricular effective refractory periods (VERPs), but did not alter action potential duration (APD), thereby reducing local critical intervals for re-excitation given by APD₉₀ − VERP. Bipolar electrogram recordings showed that 2 mM heptanol increased electrogram duration (EGD) and ratios of EGDs obtained at the longest to those obtained at the shortest S1S2 intervals studied during PES, suggesting increased dispersion of conduction velocities. These findings show, for the first time in the mouse heart, that slowed conduction induces reversible arrhythmogenic effects despite repolarization abnormalities expected to reduce arrhythmogenicity.     

Empirical correlation of triggered activity and spatial and temporal re-entrant substrates with arrhythmogenicity in a murine model for Jervell and Lange-Nielsen syndrome

KCNE1 encodes the β-subunit of the slow component of the delayed rectifier K⁺ current. The Jervell and Lange-Nielsen syndrome is characterized by sensorineural deafness, prolonged QT intervals, and ventricular arrhythmogenicity. Loss-of-function mutations in KCNE1 are implicated in the JLN2 subtype. We recorded left ventricular epicardial and endocardial monophasic action potentials (MAPs) in intact, Langendorff-perfused mouse hearts. KCNE1 ^−/− but not wild-type (WT) hearts showed not only triggered activity and spontaneous ventricular tachycardia (VT), but also VT provoked by programmed electrical stimulation. The presence or absence of VT was related to the following set of criteria for re-entrant excitation for the first time in KCNE1 ^−/− hearts: Quantification of APD₉₀, the MAP duration at 90% repolarization, demonstrated alterations in (1) the difference, ∆APD₉₀, between endocardial and epicardial APD₉₀ and (2) critical intervals for local re-excitation, given by differences between APD₉₀ and ventricular effective refractory period, reflecting spatial re-entrant substrate. Temporal re-entrant substrate was reflected in (3) increased APD₉₀ alternans, through a range of pacing rates, and (4) steeper epicardial and endocardial APD₉₀ restitution curves determined with a dynamic pacing protocol. (5) Nicorandil (20 μM) rescued spontaneous and provoked arrhythmogenic phenomena in KCNE1 ^−/− hearts. WTs remained nonarrhythmogenic. Nicorandil correspondingly restored parameters representing re-entrant criteria in KCNE1 ^−/− hearts toward values found in untreated WTs. It shifted such values in WT hearts in similar directions. Together, these findings directly implicate triggered electrical activity and spatial and temporal re-entrant mechanisms in the arrhythmogenesis observed in KCNE1 ^−/− hearts.     

Identification and functional characterization of a novel KCNE2 (MiRP1) mutation that alters HERG channel kinetics

Long-QT syndrome (LQTS) may cause syncope and sudden death due to cardiac tachyarrhythmia. Chromosome 7-linked LQTS (LQT2) has been correlated with mutations in the human ether-a-go-go-related gene (HERG). HERG forms voltage-gated K channels that may be associated with Mink-related peptide 1 (MiRP1), an auxiliary beta-subunit. The channels mediate currents that resemble native I(Kr). Mutations in the KCNE2 gene encoding MiRP1 may also cause LQTS. In this study, the frequency of mutations in KCNE2 of 150 unrelated LQTS patients without known genotype and of 100 controls was analyzed using single-strand conformation polymorphism analysis and direct sequencing. We identified a novel missense mutation, V65 M, in the KCNE2 gene of a 17-year-old female with syncope and LQTS. Expression studies in Chinese hamster ovary cells revealed that mutant and wild-type MiRP1 co-localized with HERG subunits and formed functional channels. However, mutant HERG/MiRP1(V65M) channels mediated currents with an accelerated inactivation time course compared with wild-type channels. The accelerated inactivation time course of HERG/MiRP1(V65M) channels may decrease I(Kr) current density of myocardial cells, thereby impairing the ability of myocytes to repolarize in response to sudden membrane depolarizations such as extrasystoles.     

Refractory dispersion promotes conduction disturbance and arrhythmias in a Scn5a +/? mouse model

Accentuated right ventricular (RV) gradients in action potential duration (APD) have been implicated in the arrhythmogenicity observed in Brugada syndrome in studies assuming that ventricular effective refractory periods (VERPs) vary in concert with APDs. The present experiments use a genetically modified mouse model to explore spatial heterogeneities in VERP that in turn might affect conduction velocity, thereby causing arrhythmias. Activation latencies, APDs and VERPs recorded during programmed S1S2 protocols were compared in RV and left ventricular (LV) epicardia and endocardia of Langendorff-perfused wild-type (WT) and Scn5a (+/-) hearts. Scn5a (+/-) and WT hearts showed similar patterns of shorter VERPs in RV than LV epicardia, and in epicardia than endocardia. However, Scn5a (+/-) hearts showed longer VERPs, despite shorter APD(90)s, than WT in all regions examined. The pro- and anti-arrhythmic agents flecainide and quinidine increased regional VERPs despite respectively decreasing and increasing the corresponding APD(90)s particularly in Scn5a (+/-) RV epicardia. In contrast, Scn5a (+/-) hearts showed greater VERP gradients between neighbouring regions, particularly RV transmural gradients, than WT (9.1 ± 1.1 vs. 5.7 ± 0.5 ms, p < 0.05, n = 12). Flecainide increased (to 21 ± 0.9 ms, p < 0.05, n = 6) but quinidine decreased (to 4.5 ± 0.5 ms, p < 0.05, n = 6) these gradients, particularly across the Scn5a (+/-) RV. Finally, Scn5a (+/-) hearts showed greater conduction slowing than WT following S2 stimuli, particularly with flecainide administration. Rather than arrhythmogenesis resulting from increased transmural repolarization gradients in an early, phase 2, reentrant excitation mechanism, the present findings implicate RV VERP gradients in potential reentrant mechanisms involving impulse conduction slowed by partial refractoriness.     

Emerging patterns of cardiac conduction in the chick embryo: waveform analysis with photodiode array-based optical imaging.

Major difficulties investigating the developing cardiac conduction system stem from that the embryonic heart is extremely small (< 2 mm) and cardiac activation is relatively rapid (< 8 msec). The objective of this study was to investigate the electrophysiology of the embryonic chick cardiac conduction system at periseptation stages with a photodiode array-based detection method of optical mapping capable of high spatial and temporal resolution. Previous work indicated that, in chicken embryos, a switch occurs in ventricular activation pattern from immature base-to-apex to mature apex-to-base pattern at the time of ventricular septation. It was our aim to map activation in more detail to identify the active pathway or pathways of atrioventricular conduction at these particular stages. Analysis of preseptated hearts (n = 10) showed that the latest atrial activation took place just above the site of the earliest ventricular activation at the ventral left ventricular base. Analysis of postseptated hearts (n = 11) showed apex-to-base conduction consistent with activation through the maturing His-Purkinje system. Evaluation of hearts during septation revealed a gradual transition of ventricular activation patterns rather than an abrupt "switch." External pacing of preseptated hearts revealed significant slowing of interventricular conduction compared with spontaneous beats (spontaneous, 61.7 cm/sec +/- 9 cm/sec vs. paced, 36.5 cm/sec +/- 10 cm/sec). The more detailed mapping revealed that, before septation, the pattern of activation of the ventricular myocardium is consistent with direct atrial-ventricular myocardial connections at the left lateral atrioventricular junction; however, functional evidence for a preferential conduction pathway within the ventricles was present before septation.     

Nifedipine and diltiazem suppress ventricular arrhythmogenesis and calcium release in mouse hearts

Ventricular arrhythmogenesis leading to sudden cardiac death remains responsible for significant mortality in conditions such as cardiac failure and the long-QT syndrome (LQTS). Arrhythmias may be accentuated by -adrenergic stimulation and, accordingly, the present study explored the possible effects of -adrenergic stimulation and L-type Ca²⁺ channel blockade on ventricular arrhythmogenesis and Ca²⁺ handling using the mouse heart as an experimental system. Studies in whole, Langendorff-perfused hearts using programmed electrical stimulation protocols adapted from clinical practice demonstrated sustained ventricular tachycardia following addition of 0.1 µM isoprenaline (n=15), whilst no arrhythmias were observed in the absence of the drug (n=15). Arrhythmias were suppressed by nifedipine or diltiazem pre-treatment (both 1 µM) (n=8 and 4 respectively) and were also induced by elevating external [Ca²⁺] (n=3). At the cellular level, 0.1 µM isoprenaline significantly increased normalized fluorescence (F/F₀) in field-stimulated fluo-3-loaded mouse ventricular myocytes imaged using confocal microscopy, reflecting increases in sarcoplasmic reticulum Ca²⁺ release (n=8). Elevated external [Ca²⁺] also increased F/F₀ (n=4) whilst 0.1 µM nifedipine or 0.1 µM diltiazem significantly decreased F/F₀ (n=13 and 6 respectively). Pre-treatment with 0.1 µM nifedipine or 0.1 µM diltiazem suppressed the increases in F/F₀ induced by 0.1 µM isoprenaline alone (n=14 and 6 respectively). The findings thus paralleled suppression of isoprenaline-induced arrhythmias seen with nifedipine or diltiazem at the whole-heart level. Taken together, the findings may have implications for the use of L-type Ca²⁺ channel blockade in conditions associated with -adrenergically driven ventricular arrhythmias such as cardiac failure and LQTS.     

Novel KCNE3 mutation reduces repolarizing potassium current and associated with long QT syndrome

Long QT syndrome (LQTS) is an inherited disease involving mutations in the genes encoding a number of cardiac ion channels and a membrane adaptor protein. Among the genes that are responsible for LQTS, KCNE1 and KCNE2 are members of the KCNE family of genes, and function as ancillary subunits of Kv channels. The third KCNE gene, KCNE3, is expressed in cardiac myocytes and interacts with KCNQ1 to change the channel properties. However, KCNE3 has never been linked to LQTS. To investigate the association between KCNE3 and LQTS, we conducted a genetic screening of KCNE3 mutations and single nucleotide polymorphisms (SNPs) in 485 Japanese LQTS probands using DHPLC‐WAVE system and direct sequencing. Consequently, we identified two KCNE3 missense mutations, located in the N‐ and C‐terminal domains. The functional effects of these mutations were examined by heterologous expression systems using CHO cells stably expressing KCNQ1. One mutation, p.R99λH was identified in a 76‐year‐old woman who suffered torsades de pointes (TdP) after administration of disopyramide. Another mutation, p.T4A was identified in a 16‐year‐old boy and 67‐year‐old woman. Although the boy carried another KCNH2 mutation, he was asymptomatic. On the other hand, the woman suffered from hypokalemia‐induced TdP. In a series of electrophysiological analyses, the KCNQ1(Q1)+KCNE3(E3)‐R99λH channel significantly reduced outward current compared to Q1+E3‐WT, though the current density of the Q1+E3‐T4A channel displayed no statistical significance. This is the first report of KCNE3 mutations associated with LQTS. Screening for variants in the KCNE3 gene is of clinical importance for LQTS patients. Hum Mutat 30, 557–563, 2009. © 2009 Wiley‐Liss, Inc.     

Criteria for arrhythmogenicity in genetically-modified Langendorff-perfused murine hearts modelling the congenital long QT syndrome type 3 and the Brugada syndrome

The experiments investigated the applicability of two established criteria for arrhythmogenicity in Scn5a+/Δ and Scn5a+/− murine hearts modelling the congenital long QT syndrome type 3 (LQT3) and the Brugada syndrome (BrS). Monophasic action potentials (APs) recorded during extrasystolic stimulation procedures from Langendorff-perfused control hearts and hearts treated with flecainide (1 μM) or quinidine (1 or 10 μM) demonstrated that both agents were pro-arrhythmic in wild-type (WT) hearts, quinidine was pro-arrhythmic in Scn5a+/Δ hearts, and that flecainide was pro-arrhythmic whereas quinidine was anti-arrhythmic in Scn5a+/− hearts, confirming clinical findings. Statistical analysis confirmed a quadratic relationship between epicardial and endocardial AP durations (APDs) in WT control hearts. However, comparisons between plots of epicardial against endocardial APDs and this reference curve failed to correlate with arrhythmogenicity. Restitution curves, relating APD to diastolic interval (DI), were then constructed for the first time in a murine system and mono-exponential growth functions fitted to these curves. Significant (P < 0.05) alterations in the DI at which slopes equalled unity, an established indicator of arrhythmogenicity, now successfully predicted the presence or absence of arrhythmogenicity in all cases. We thus associate changes in the slopes of restitution curves with arrhythmogenicity in models of LQT3 and BrS.     

The role of KCNQ1/KCNE1 K+ channels in intestine and pancreas: lessons from the KCNE1 knockout mouse

KCNE1 (IsK, minK) co-assembles with KCNQ1 (KvLQT1) to form voltage-dependent K(+) channels. Both KCNQ1 and KCNE1 are expressed in epithelial cells of gut and exocrine pancreas. We examined the role of KCNQ1/KCNE1 in Cl(-) secretion in small and large intestine and exocrine pancreas using the KCNE1 knockout mouse. Immunofluorescence revealed a similar basolateral localization of KCNQ1 in jejunum and colon of KCNE1 wild-type and knockout mice. Electrogenic Cl(-) secretion in the colon was not affected by gene disruption of KCNE1; in jejunum forskolin-induced short-circuit current was some 40% smaller but without being significantly different. Inhibition of KCNQ1 channels by 293B (IC(50) 1 micromol l(-1)) and by IKS224 (IC(50) 14 nmol l(-1)) strongly diminished intestinal Cl(-) secretion. In exocrine pancreas of wild-type mice, KCNQ1 was predominantly located at the basolateral membrane. In KCNE1 knockout mice, however, the basolateral staining was less pronounced and the distribution of secretory granules was irregular. A slowly activating and 293B-sensitive K(+) current was activated via cholinergic stimulation in pancreatic acinar cells of wild-type mice. In KCNE1 knockout mice this K(+) current was strongly reduced. In conclusion intestinal Cl(-) secretion is independent from KCNE1 but requires KCNQ1. In mouse pancreatic acini KCNQ1 probably co-assembled with KCNE1 leads to a voltage-dependent K(+) current that might be of importance for electrolyte and enzyme secretion.     

Spectrum of pathogenic mutations and associated polymorphisms in a cohort of 44 unrelated patients with long QT syndrome

Long QT syndrome (LQTS) is a rare and clinically heterogeneous inherited disorder characterized by a long QT interval on the electrocardiogram, increased risk of syncope and sudden death caused by arrhythmias. This syndrome is mostly caused by mutations in genes encoding various cardiac ion channels. The clinical heterogeneity is usually attributed to variable penetrance. One of the reasons for this variability in expression could be the coexistence of common single nucleotide polymorphisms (SNPs) on LQTS-causing genes and/or unknown genes. Some synonymous and nonsynonymous exonic SNPs identified in LQTS-causing genes may have an effect on the cardiac repolarization process and modulate the clinical expression of a latent LQTS pathogenic mutation. We report the molecular pattern of 44 unrelated patients with LQTS using denaturing high-performance liquid chromatography analysis of the KCNQ1, KCNH2, SCN5A, KCNE1 and KCNE2 genes. Forty-five disease-causing mutations (including 24 novel ones) were identified in this cohort. Most of our patients (84%) showed complex molecular pattern with one mutation (and even two for four patients) associated with several SNPs located in several LQTS genes.     

Reduced urinary bladder afferent conduction velocities in streptozocin diabetic rats.

Previous experiments in our laboratory have described the method used to measure the conduction velocity distribution of a selected group of fibers (Brain Res., 520 (1990) 83-89). We have applied this technique to the 2 month streptozotocin-diabetic rat. Glycosylated hemoglobin values measured at the time of death were 17.19 +/- 4.74% (diabetic, n = 8) and 4.07 +/- 0.74% (controls, n = 6). Diabetic bladders were thicker and heavier. The wet weights were 0.50 +/- 0.11 g (diabetic, n = 7) and 0.16 +/- 0.01 g (controls, n = 6). The conduction velocities of a total of 151 and 86 single afferent fibers were measured in the diabetic and control animals respectively. The conduction velocity distribution of the diabetics showed a shift towards slower speeds when compared to controls. The mean conduction velocities were 1.70 m/s for diabetics and 2.84 m/s for controls. The percent of units with conduction velocities greater than 2.5 m/s was 17.2 for diabetics and 36.0 for controls. This experiment demonstrates, for the first time, that diabetes causes a significant reduction of afferent conduction velocities in a functionally well-defined system.     

LQTS gene LOVD database

The Long QT Syndrome (LQTS) is a group of genetically heterogeneous disorders that predisposes young individuals to ventricular arrhythmias and sudden death. LQTS is mainly caused by mutations in genes encoding subunits of cardiac ion channels (KCNQ1, KCNH2, SCN5A, KCNE1, and KCNE2). Many other genes involved in LQTS have been described recently (KCNJ2, AKAP9, ANK2, CACNA1C, SCNA4B, SNTA1, and CAV3). We created an online database ( http://www.genomed.org/LOVD/introduction.html ) that provides information on variants in LQTS‐associated genes. As of February 2010, the database contains 1738 unique variants in 12 genes. A total of 950 variants are considered pathogenic, 265 are possible pathogenic, 131 are unknown/unclassified, and 292 have no known pathogenicity. In addition to these mutations collected from published literature, we also submitted information on gene variants, including one possible novel pathogenic mutation in the KCNH2 splice site found in ten Chinese families with documented arrhythmias. The remote user is able to search the data and is encouraged to submit new mutations into the database. The LQTS database will become a powerful tool for both researchers and clinicians. © 2010 Wiley‐Liss, Inc.     

DHPLC analysis of potassium ion channel genes in congenital long QT syndrome

Congenital long QT syndrome (LQTS) is electrocardiographically characterized by a prolonged QT interval and polymorphic ventricular arrhythmias (torsade de pointes). As a result of these arrhythmias, patients suffer from recurrent syncopes, seizures, or sudden death as the most dramatic event. Mutations in five genes, encoding cardiac ion channels, have been identified in LQTS. Two potassium-channel genes, KCNQ1 (LQT1) and KCNH2 (LQT2 or HERG), are frequently involved in LQTS. Potassium-channel defects account for approximately 50-60% of LQTS. As patients benefit from preventive medication, early detection of a genetic defect is desired to identify the family members at risk. Speed and sensitivity of mutation detection was improved by applying the denaturing high performance liquid chromatography (DHPLC) technique for analysis of the entire KCNQ1 and KCNH2 genes and the protein encoding part of the KCNE1 and KCNE2 genes. By using this methodology, seven missense mutations in the KCNQ1 gene and nine mutations (four missense, two nonsense, one insertion, and two deletions) in the KCNH2 gene have been identified in a total number of 32 index patients diagnosed with LQTS syndrome. We conclude that this method is suitable for rapid identification of LQT gene defects due to the combination of automation, high throughput, sensitivity, and short time of analysis.     

卡维地洛抑制大鼠起搏心肌细胞钙库超载诱导的钙释放*

 目的：探讨非选择性β受体阻滞剂卡维地洛对起搏心肌细胞钙库超载诱导钙释放（SOICR）的作用及其机制。方法：电刺激起搏大鼠单个心肌细胞并灌注异丙肾上腺素及咖啡因诱导钙库钙超载,从而触发肌浆网钙释放通道（兰尼碱受体2,RyR2）舒张期开放引起SOICR。应用钙离子荧光成像技术记录胞内钙浓度的实时变化。实验分为对照组、卡维地洛组、美托洛尔组、酚妥拉明组和硝苯地平组。结果：(1) 与基线刺激时比较,对照组细胞灌注异丙肾上腺素和咖啡因后,起搏细胞钙瞬变振幅显著增高（P<0.01）,SOICR的发生率显著增加（P<0.01）。(2) 在1~4 Hz起搏频率下卡维地洛组细胞SOICR发生率分别为2.00%、6.00%、10.00%和16.00%,均显著低于对照组（分别为43.59%、74.36%、87.18%和89.74%,均P<0.01）;卡维地洛对心肌细胞SOICR的抑制在不同起搏频率下无明显差异（P>0.05）。酚妥拉明、美托洛尔和硝苯地平组细胞与对照组细胞比较,SOICR发生率无差异（均P>0.05）。(3) 起搏细胞钙瞬变振幅的比较,各组间相比未见明显差异（P>0.05）;咖啡因峰值估测钙库钙总量的比较,各组间也无明显差异（P>0.05）。结论：卡维地洛可明显抑制起搏心肌细胞SOICR的发生,其作用机制可能是直接抑制RyR2的自发性开放而非源于对α1、β1受体和L型钙通道的阻滞作用。     

Decreased KCNE2 expression participates in the development of cardiac hypertrophy

AIM: To investigate whether KCNE2 participates in the development of pathological hypertrophy. METHODS: Bidirectional manipulations of KCNE2 expression were performed by adenoviral overexpression of KCNE2 or knockdown of KCNE2 with RNA interference in PE-induced neonatal rat ventricular myocytes. Then overexpression of KCNE2 in mouse model of left ventricular hypertrophy induced by transverse aortic constriction( TAC) by ultrasound microbubble-mediated gene transfer were used to detect the therapeutic function of KCNE2 in the development of hypertrophy. RESULTS: KCNE2 expression was significantly decreased in PE-induced hypertrophic cardiomyocytes and in hypertrophic hearts produced by TAC. Knockdown of KCNE2 in cardiomyocytes reproduced hypertrophy,whereas overexpression of KCNE2 attenuated PE-induced cardiomyocyte hypertrophy. Knockdown of KCNE2 increased calcineurin activity and nuclear NFAT protein level,and pretreatment with nifedipine or FK506 attenuated decreased KCNE2-induced cardiomyocyte hypertrophy. Overexpression of KCNE2 in heart by ultrasound microbubble-mediated gene transfer suppressed the development of hypertrophy and activation of calcineurin-NFAT and MAPK pathways in TAC mice. CONCLUSION: These findings demonstrate that cardiac KCNE2 expression is decreased and contributes to the development of hypertrophy via activation of calcineurin-NFAT and MAPK pathways.     

Long-range projections of Adelta primary afferents in the Lissauer tract of the rat

Electrical microstimulation has been used to activate fine myelinated primary afferents running within the Lissauer tract. Stimulation of the tract at the L2/L3 border produced antidromic volleys which were recorded on the dorsal roots of more caudal spinal segments. Antidromic volleys were present in all cases for roots as far caudal as the S2 segment (L3, n=12; L4, n=6; L5, n=6; L6, n=9; S1, n=3; S2, n=6; observations in a total of 15 rats). These fibres were collaterals of primary afferents with conduction velocities in the dorsal root of up to 17.3+/-2.3 ms(-1) (mean+/-S.D., n=6; range 14-20 ms(-1)). Conduction velocities within the Lissauer tract were slower; the fastest contributing fibres had conduction velocities of 9.2+/-2.2 ms(-1) (range 6-12 ms(-1)). Lesions of the Lissauer tract caudal to the stimulation site abolished the volleys on roots lying caudal to the lesion. Most previous works have suggested that primary afferents project in the Lissauer tract for only one or two spinal segments. The present study shows that some fibres project rostrally for up to seven spinal segments (L2-S2).     

Regulation of KCNE1-dependent K(+) current by the serum and glucocorticoid-inducible kinase (SGK) isoforms

The slowly activating K(+) channel subunit KCNE1 is expressed in a variety of tissues including proximal renal tubules, cardiac myocytes and stria vascularis of inner ear. The present study has been performed to explore whether the serum- and glucocorticoid-inducible kinase family members SGK1, SGK2, or SGK3 and/or protein kinase B (PKB) influence K(+) channel activity in Xenopus oocytes expressing KCNE1. cRNA encoding KCNE1 was injected with or without cRNA encoding wild-type SGK1, constitutively active (S422D)SGK1, inactive (K127 N)SGK1, wild-type SGK2, wild-type SGK3 or constitutively active (T308D,S473D)PKB. In oocytes injected with KCNE1 cRNA but not in water-injected oocytes a depolarization from -80 mV to -10 mV led to the appearance of a slowly activating K(+) current. Coexpression of SGK1,( S422D)SGK1, SGK2, SGK3 or (T308D,S473D)PKB but not (K127 N)SGK1 significantly stimulated KCNE1-induced current. The effect did not depend on Na(+)/K(+)-ATPase activity. KCNE1-induced current was markedly upregulated by coexpression of KCNQ1 and further increased by additional expression of (S422D)SGK1, SGK2, SGK3 or (T308D,S473D)PKB. In conclusion, all three members of the SGK family of kinases SGK1-3 and protein kinase B stimulate the slowly activating K(+) channel KCNE1/KCNQ1. The kinases may thus participate in the regulation of KCNE1-dependent transport and excitability.     

Non-uniformity of two-dimensional myocardial deformation in response to chronotropic and inotropic stimulation in cats

The degree of uniformity of myocardial deformation for cross-oriented segments in the anterior wall of the left ventricle is influenced both by loading conditions and by infusion of isoprenaline. The aim of this study was to clarify the chronotropic influence (atrial pacing) compared to the combined inotropic and chronotropic effects of isoprenaline on uniformity of contraction. In eight open-chest pentobarbitone-anaesthetized cats segment performance was measured by orthogonal sonomicrometry. Heart rate (HR) increased from control state, 199 ± 5 (mean ± SEM) beats min^-1, to 224 ± 6 and 227 ± 7 beats min^-1 during atrial pacing and isoprenaline infusion, respectively (P < 0.0005). Circumferential segment shortening remained unchanged during pacing but increased with isoprenaline (P < 0.0005). Longitudinal segments showed reduced shortening during pacing (P < 0.05), whereas shortening during isoprenaline infusion did not differ from control shortening. The ratio between shortening of longitudinal and circumferential segments, long/circ ratio, changed from 0.52 ± 0.13 in the control state to 0.36 ± 0.10 during pacing and 0.39 ± 0.11 with isoprenaline (P < 0.05). End-systolic pressure-length relations were unchanged by atrial pacing but showed leftward shifts during isoprenaline infusion. Myocardial tissue blood flow (microspheres) was unchanged during interventions. We conclude that both atrial pacing and isoprenaline infusion lead to a more non-uniform deformation in the anterior wall. However, non-uniformity during atrial pacing was primarily related to reduced longitudinal shortening whereas the non-uniformity during isoprenaline infusion was due to increased circumferential shortening.     

Attenuation of conduction delay by ischemic preconditioning reduces ischemia-induced ventricular arrhythmias

Ischemic preconditioning has been acknowledged as a powerful method of decreasing ischemic injury. However, the antiarrhythmic mechanism of ischemic preconditioning during ischemia is unclear. We studied the effects of ischemic preconditioning on arrhythmias and cardiac electrophysiology during ischemia in Langendorff rat hearts (n = 44). In the non-preconditioned group (PC(-); n = 24), the hearts underwent 5-min zero-flow global ischemia without any prior ischemic preconditioning. In the preconditioned group (PC(+); n = 20), the hearts were preconditioned by three cycles of 3-min zero-flow global ischemia and 5-min reperfusion before undergoing 5-min global ischemia. Ischemic preconditioning reduced the incidence of ischemia-induced arrhythmias (PC(-); 38.9%, PC(+): 8.3%, p < 0.05), shortened monophasic action potential duration (MAPD, P < 0.05), attenuated conduction delay (conduction time; PC(-): 234.2%, PC(+): 173.4%, P < 0.05) and increased the ventricular fibrillation threshold. Although the shortening of MAPD in PC(-) hearts was not influenced by the presence or absence of arrhythmias, conduction time prolongation at 3-min was more obvious in PC(-) hearts with arrhythmia than in PC(-) hearts without arrhythmia (PC(-) with arrhythmia: 220.2%, PC(-) without arrhythmia: 190.7%, P < 0.05). We concluded that ischemic preconditioning could protect the rat hearts from ischemia-induced arrhythmias and postulated that attenuation of conduction delay during ischemia might be an important factor in the antiarrhythmic action of ischemic preconditioning.