Departamento de Fisiología, Facultad de Medicina, Universidad Nacional Autónoma de México, México DF, México.
Kidney Int. 2011 Oct;80(8):832-40. doi: 10.1038/ki.2011.230. Epub 2011 Jul 27.
Recent studies have identified Rhesus proteins as important molecules for ammonia transport in acid-secreting intercalated cells in the distal nephron. Here, we provide evidence for an additional molecule that can mediate NH3/NH4 excretion, the subtype 2 of the hyperpolarization-activated cyclic nucleotide-gated channel family (HCN2), in collecting ducts in rat renal cortex and medulla. Chronic metabolic acidosis in rats did not alter HCN2 protein expression but downregulated the relative abundance of HCN2 mRNA. Its cDNA was identical to the homolog from the brain and the protein was post-translationally modified by N-type glycosylation. Electrophysiological recordings in Xenopus oocytes injected with HCN2 cRNA found that potassium was transported better than ammonium, each of which was transported significantly better than sodium, criteria that are compatible with a role for HCN2 in ammonium transport. In microperfused rat outer medullary collecting duct segments, the initial rate of acidification, upon exposure to a basolateral ammonium chloride pulse, was higher in intercalated than in principal cells. A specific inhibitor of HCN2 (ZD7288) decreased acidification only in intercalated cells from control rats. In rats with chronic metabolic acidosis, the rate of acidification doubled in both intercalated and principal cells; however, ZD7288 had no significant inhibitory effect. Thus, HCN2 is a basolateral ammonium transport pathway of intercalated cells and may contribute to the renal regulation of body pH under basal conditions.
最近的研究已经确定了 Rh 蛋白是远端肾单位酸化分泌闰细胞中氨转运的重要分子。在这里,我们提供了证据表明另一种可以介导 NH3/NH4 排泄的分子,即超极化激活环核苷酸门控通道家族(HCN2)的亚型 2,在大鼠肾皮质和髓质的集合管中。大鼠慢性代谢性酸中毒不会改变 HCN2 蛋白的表达,但下调了 HCN2 mRNA 的相对丰度。其 cDNA 与大脑中的同源物相同,并且蛋白质通过 N 型糖基化进行翻译后修饰。在注射了 HCN2 cRNA 的非洲爪蟾卵母细胞中的电生理记录发现,钾的转运优于铵,而每种物质的转运都明显优于钠,这与 HCN2 在铵转运中的作用一致。在微灌注的大鼠外髓集合管段中,暴露于基底外侧氯化铵脉冲时,酸化的初始速率在闰细胞中比在主细胞中更高。HCN2 的特异性抑制剂(ZD7288)仅在对照大鼠的闰细胞中降低酸化。在慢性代谢性酸中毒的大鼠中,闰细胞和主细胞的酸化速率均增加了一倍;然而,ZD7288 没有显著的抑制作用。因此,HCN2 是闰细胞的基底外侧铵转运途径,可能有助于在基础条件下调节肾脏的 pH 值。