Kassiri Zamaneh, Zobel Carsten, Nguyen The-Tin T, Molkentin Jeffery D, Backx Peter H
Department of Physiology, Heart and Stroke/Richard Lewar Center, University of Toronto, Toronto, Canada.
Circ Res. 2002 Mar 22;90(5):578-85. doi: 10.1161/01.res.0000012223.86441.a1.
Prolonged action potential duration (APD) and decreased transient outward K+ current (I(to)) as a result of decreased expression of K(v4.2) and K(v4.3) genes are commonly observed in heart disease. We found that treatment of cultured neonatal rat ventricular myocytes with Heteropoda Toxin3, a blocker of cardiac I(to), induced hypertrophy as measured using cell membrane capacitance and (3)H-leucine uptake. To dissect the role of specific I(to)-encoding genes in hypertrophy, I(to) was selectively reduced by overexpressing mutant dominant-negative (DN) transgenes. I(to) amplitude was reduced equally (by about 50%) by overexpression of DN K(v1.4) (K(v1.4)N) or DN K(v4.2) (either K(v4.2)N or K(v4.2)W362F), but only DN K(v4.2) prolonged APD duration (at 1 Hz) and induced myocyte hypertrophy. This hypertrophy was prevented by coexpressing wild-type K(v4.2) channels (K(v4.2)F) with the DN K(v4.2) genes, suggesting the hypertrophy is due to I(to) reduction and not nonspecific effects of transgene overexpression. The hypertrophy caused by reductions of K(v4.x)-based I(to) was associated with increased activity of the calcium-dependent phosphatase, calcineurin, and could be prevented by coinfection with Ad-CAIN, a specific calcineurin inhibitor. The hypertrophy and calcineurin activation induced by K(v4.2)N infection were prevented by blocking Ca2+ entry and excitability with verapamil or high [K+]o. Our studies suggest that reductions of K(v4.2/3)-based I(to) play a role in hypertrophy signaling by activation of calcineurin.
在心脏病中通常可观察到动作电位时程(APD)延长以及由于K(v4.2)和K(v4.3)基因表达降低导致的瞬时外向K +电流(I(to))减少。我们发现,用心脏I(to)阻滞剂异足蛛毒素3处理培养的新生大鼠心室肌细胞会诱导肥大,这可通过细胞膜电容和3H-亮氨酸摄取来测量。为了剖析特定I(to)编码基因在肥大中的作用,通过过表达突变显性负性(DN)转基因选择性降低I(to)。过表达DN K(v1.4)(K(v1.4)N)或DN K(v4.2)(K(v4.2)N或K(v4.2)W362F)可使I(to)幅度同等降低(约50%),但只有DN K(v4.2)延长了APD时程(在1 Hz时)并诱导心肌细胞肥大。通过将野生型K(v4.2)通道(K(v4.2)F)与DN K(v4.2)基因共表达可防止这种肥大,这表明肥大是由于I(to)降低而非转基因过表达的非特异性效应所致。基于K(v4.x)的I(to)减少所引起的肥大与钙依赖性磷酸酶钙调神经磷酸酶的活性增加有关,并且可通过与特异性钙调神经磷酸酶抑制剂Ad-CAIN共感染来预防。用维拉帕米或高[K +] o阻断Ca2 +内流和兴奋性可预防由K(v4.2)N感染诱导的肥大和钙调神经磷酸酶激活。我们的研究表明,基于K(v4.2/3)的I(to)减少通过激活钙调神经磷酸酶在肥大信号传导中起作用。