Jiangsu Province Key Laboratory of Anesthesiology, Xuzhou Medical College, Xuzhou 221002, China;Department of Pharmacology, New York Medical College, Valhalla, NY 10595; and.
Department of Pharmacology, New York Medical College, Valhalla, NY 10595; and.
Proc Natl Acad Sci U S A. 2014 Aug 12;111(32):11864-9. doi: 10.1073/pnas.1411705111. Epub 2014 Jul 28.
The renal phenotype induced by loss-of-function mutations of inwardly rectifying potassium channel (Kir), Kcnj10 (Kir4.1), includes salt wasting, hypomagnesemia, metabolic alkalosis and hypokalemia. However, the mechanism by which Kir.4.1 mutations cause the tubulopathy is not completely understood. Here we demonstrate that Kcnj10 is a main contributor to the basolateral K conductance in the early distal convoluted tubule (DCT1) and determines the expression of the apical Na-Cl cotransporter (NCC) in the DCT. Immunostaining demonstrated Kcnj10 and Kcnj16 were expressed in the basolateral membrane of DCT, and patch-clamp studies detected a 40-pS K channel in the basolateral membrane of the DCT1 of p8/p10 wild-type Kcnj10(+/+) mice (WT). This 40-pS K channel is absent in homozygous Kcnj10(-/-) (knockout) mice. The disruption of Kcnj10 almost completely eliminated the basolateral K conductance and decreased the negativity of the cell membrane potential in DCT1. Moreover, the lack of Kcnj10 decreased the basolateral Cl conductance, inhibited the expression of Ste20-related proline-alanine-rich kinase and diminished the apical NCC expression in DCT. We conclude that Kcnj10 plays a dominant role in determining the basolateral K conductance and membrane potential of DCT1 and that the basolateral K channel activity in the DCT determines the apical NCC expression possibly through a Ste20-related proline-alanine-rich kinase-dependent mechanism.
由内向整流钾通道(Kir)、Kcnj10(Kir4.1)功能丧失突变引起的肾表型包括盐耗、低镁血症、代谢性碱中毒和低钾血症。然而,Kir.4.1 突变导致小管病的机制尚不完全清楚。在这里,我们证明 Kcnj10 是早期远曲小管(DCT1)基底外侧 K 电导的主要贡献者,并决定了 DCT 中顶端 Na-Cl 共转运蛋白(NCC)的表达。免疫染色显示 Kcnj10 和 Kcnj16 表达在 DCT 的基底外侧膜上,膜片钳研究检测到 p8/p10 野生型 Kcnj10(+/+)(WT)小鼠 DCT1 的基底外侧膜上存在 40-pS K 通道。这种 40-pS K 通道在纯合 Kcnj10(-/-)(敲除)小鼠中不存在。Kcnj10 的破坏几乎完全消除了 DCT1 中的基底外侧 K 电导并降低了细胞膜电位的负性。此外,缺乏 Kcnj10 降低了基底外侧 Cl 电导,抑制了 Ste20 相关脯氨酸-丙氨酸丰富激酶的表达,并减少了 DCT 中的顶端 NCC 表达。我们得出结论,Kcnj10 在决定 DCT1 的基底外侧 K 电导和膜电位方面起着主导作用,DCT 中的基底外侧 K 通道活性可能通过 Ste20 相关脯氨酸-丙氨酸丰富激酶依赖性机制决定顶端 NCC 的表达。