Department of Anatomy, Ewha Womans University, Seoul, Korea.
Am J Physiol Renal Physiol. 2013 Apr 1;304(7):F972-81. doi: 10.1152/ajprenal.00550.2012. Epub 2013 Jan 16.
The ammonia transporter family member, Rh B Glycoprotein (RhBG/Rhbg), is essential for ammonia transport by the rodent kidney, but in the human kidney mRNA but not protein expression has been reported. Because ammonia transport is fundamental for acid-base homeostasis, the current study addressed RhBG expression in the human kidney. Two distinct RhBG mRNA sequences have been reported, with different numbers of consecutive cytosines at nt1265 and thus encoding different carboxy-tails. Sequencing the region of difference in both human kidney and liver mRNA showed eight sequential cytosines, not seven as in some reports. Knowing the correct mRNA sequence for RhBG, we then assessed RhBG protein expression using antibodies against the correct amino acid sequence. Immunoblot analysis demonstrated RhBG protein expression in human kidney and immunohistochemistry identified basolateral RhBG in connecting segment (CNT) and the cortical and outer medullary collecting ducts. Colocalization of RhBG with multiple cell-specific markers demonstrated that that CNT cells and collecting duct type A intercalated cells express high levels of RhBG, and type B intercalated cells and principal cells do not express detectable RhBG. Thus, these studies identify the correct mRNA and thus protein sequence for human RhBG and show that the human kidney expresses basolateral RhBG protein in CNT, type A intercalated cells, and non-A, non-B cells. We conclude that RhBG can mediate an important role in human renal ammonia transport.
氨转运体家族成员 RhB 糖蛋白(RhBG/Rhbg)对于啮齿动物肾脏中的氨转运至关重要,但在人类肾脏中仅报道了其 mRNA 表达,而未检测到蛋白表达。由于氨转运对于酸碱平衡至关重要,因此本研究旨在探讨人类肾脏中 RhBG 的表达情况。目前已报道了两种不同的 RhBG mRNA 序列,其在 nt1265 处的连续胞嘧啶数不同,因此编码的羧基尾部也不同。对人肾脏和肝脏 mRNA 差异区域进行测序,结果显示有 8 个连续的胞嘧啶,而不是一些报道中提到的 7 个。了解了 RhBG 的正确 mRNA 序列后,我们使用针对正确氨基酸序列的抗体来评估 RhBG 蛋白的表达情况。免疫印迹分析显示 RhBG 蛋白在人肾脏中表达,免疫组织化学鉴定了连接段(CNT)和皮质及外髓集合管中的 RhBG 基底外侧表达。RhBG 与多种细胞特异性标志物的共定位表明,CNT 细胞和集合管 A 型闰细胞高表达 RhBG,而 B 型闰细胞和主细胞则不表达可检测到的 RhBG。因此,这些研究确定了人类 RhBG 的正确 mRNA 序列和蛋白序列,并表明人肾脏中的 RhBG 蛋白在 CNT、A 型闰细胞和非 A、非 B 细胞中表达于基底外侧。我们得出结论,RhBG 可以在人类肾脏的氨转运中发挥重要作用。