Nakamura T Y, Coetzee W A, Vega-Saenz De Miera E, Artman M, Rudy B
Department of Pediatrics, New York University Medical Center, New York 10016, USA.
Am J Physiol. 1997 Oct;273(4):H1775-86. doi: 10.1152/ajpheart.1997.273.4.H1775.
Current evidence suggests that members of the Kv4 subfamily may encode native cardiac transient outward current (I(to)). Antisense hybrid-arrest with oligonucleotides targeted to Kv4 mRNAs specifically inhibited rat ventricular I(to), supporting this hypothesis. To determine whether protein kinase C (PKC) affects I(to) by an action on these molecular components, we compared the effects of PKC activation on Kv4.2 and Kv4.3 currents expressed in Xenopus oocytes and rat ventricular I(to). Phorbol 12-myristate 13-acetate (PMA) suppressed both Kv4.2 and Kv4.3 currents as well as native I(to), but not after preincubation with PKC inhibitors (e.g., chelerythrine). An inactive stereoisomer of PMA had no effect. Phenylephrine or carbachol inhibited Kv4 currents only when coexpressed, respectively, with alpha1C-adrenergic or M1 muscarinic receptors (this inhibition was also prevented by chelerythrine). The voltage dependence and inactivation kinetics of Kv4.2 were unchanged by PKC, but small effects on the rates of inactivation and recovery from inactivation of native I(to) were observed. Thus Kv4.2 and Kv4.3 proteins are important subunits of native rat ventricular I(to), and PKC appears to reduce this current by affecting the molecular components of the channels mediating I(to).
目前的证据表明,Kv4亚家族成员可能编码天然心脏瞬时外向电流(I(to))。用针对Kv4 mRNA的寡核苷酸进行反义杂交阻滞可特异性抑制大鼠心室I(to),支持了这一假说。为了确定蛋白激酶C(PKC)是否通过作用于这些分子成分来影响I(to),我们比较了PKC激活对非洲爪蟾卵母细胞中表达的Kv4.2和Kv4.3电流以及大鼠心室I(to)的影响。佛波醇12-肉豆蔻酸酯13-乙酸酯(PMA)抑制了Kv4.2和Kv4.3电流以及天然I(to),但在用PKC抑制剂(如白屈菜红碱)预孵育后则无此作用。PMA的无活性立体异构体没有影响。去氧肾上腺素或卡巴胆碱仅在分别与α1C-肾上腺素能或M1毒蕈碱受体共表达时才抑制Kv4电流(这种抑制也可被白屈菜红碱阻止)。PKC对Kv4.2的电压依赖性和失活动力学没有影响,但观察到对天然I(to)的失活速率和从失活状态恢复的速率有微小影响。因此,Kv4.2和Kv4.3蛋白是天然大鼠心室I(to)的重要亚基,PKC似乎通过影响介导I(to)的通道的分子成分来降低这种电流。