Background-Increased susceptibility to dilated cardiomyopathy has been observed in patients carrying mutations in the SCN5A gene, but the underlying mechanism remains unclear. In this study, we identified and characterized, both in vitro and clinically, an SCN5A mutation associated with familial progressive atrioventricular block of adult onset and dilated cardiomyopathy in a Chinese family. Methods and Results-Among 32 family members, 5 were initially diagnosed with atrioventricular block after age 30; 4 were studied, 3 of whom later developed dilated cardiomyopathy. We found a heterozygous single-nucleotide mutation resulting in an amino acid substitution (A1180V) in all studied patients and in 6 other younger unaffected members but not in 200 control chromosomes. When expressed with the ␤1 subunit, the mutated channels exhibited a Ϫ4.5-mV shift of inactivation with slower recovery leading to a rate-dependent Na ϩ current reduction and a moderate increase in late Na ϩ current. Clinical study revealed that although QRS duration decreased with increasing heart rate in noncarrier family members, this change was blunted in unaffected carriers whose ECG and heart function were normal. Resting corrected QT interval of unaffected carriers was significantly longer than that of noncarriers, even though it was still within the normal range. Conclusions-A1180V expresses a mild Na ϩ channel phenotype in vitro and a corresponding clinical phenotype in unaffected mutation carriers, implying that A1180V caused structural heart disease in affected carriers by disturbing Na

X-ray structures of the bacterial K ؉ channel KcsA have led to unparalleled progress in our understanding of ion channel structures. The KcsA channel has therefore been a prototypic model used to study the structural basis of ion channel function, including the gating mechanism. This channel was previously found to close at near-neutral intracellular pH (pHi) and to open at acidic pHi. Here, we report the presence of a previously unknown channel inactivation process that occurs after the KcsA channel is activated. In our experiments, mammalian cells transfected with a codonoptimized synthetic gene encoding the KcsA protein expressed K ؉ -selective channels that activated in response to a decrease in pHi. Using patch-clamp and rapid solution exchange techniques, we observed that the KcsA channels inactivated within hundreds of milliseconds after channel activation. At all tested pHs, inactivation always accompanied activation, and it was profoundly accelerated in the same pH range at which activation increased steeply. Recovery from inactivation was observed, and its extent depended on the pHi and the amount of time that the channel was inactive. KcsA channel inactivation can be described by a kinetic model in which pH i controls inactivation through pH-dependent activation. This heretofore-undocumented inactivation process increases the complexity of KcsA channel function, but it also offers a potential model for studying the structural correspondence of ion channel inactivation.gating ͉ ion channel inactivation ͉ recovery from inactivation ͉ synthetic gene ͉ model

SUMMARY The burbot (Lota lota) is a cold stenothermic fish species whose heart is adapted to function in the cold. In this study we use whole-cell voltage-clamp techniques to characterize the electrophysiological properties of burbot ventricular myocytes and to test the hypothesis that changes in membrane currents and intracellular Ca2+ cycling associated cold-acclimation in other fish species are routine for stenothermic cold-adapted species. Experiments were performed at 4°C, which is the body temperature of burbot for most of the year, and after myocytes were acutely warmed to 11°C, which is in the upper range of temperatures experienced by burbot in nature. Results on K+ channels support our hypothesis as the relative density of K-channel conductances in the burbot heart are similar to those found for cold-acclimated cold-active fish species. IK1 conductance was small (39.2±5.4 pS pF-1 at 4°C and 71.4±1.7 pS pF-1 at 11°C)and IKr was large (199±27 pS pF-1 at 4°C and 320.3±8 pS pF-1 at 11°C) in burbot ventricular myocytes. We found high Na+-Ca2+ exchange(NCX) activity (35.9±6.3 pS pF-1 at 4°C and 58.6±8.4 pS pF-1 at 11°C between -40 and 20 mV),suggesting that it may be the primary pathway for sarcolemmal (SL)Ca2+ influx in this species. In contrast, the density(ICa, 0.81±0.13 pA pF-1 at 4°C, and 1.35±0.18 pA pF-1 at 11°C) and the charge(QCa, 0.24±0.043 pC pF-1 at 4°C and 0.21±0.034 pC pF-1 at 11°C) carried by the l-type Ca2+ current was small. Our results on sarcolemmal ion currents in burbot ventricular myocytes suggest that cold stenothermy and compensative cold-acclimation involve many of the same subcellular adaptations that culminate in enhanced excitability in the cold.