Quentin Fabienne, Eladari Dominique, Cheval Lydie, Lopez Claude, Goossens Dominique, Colin Yves, Cartron Jean-Pierre, Paillard Michel, Chambrey Régine
Institut National de la Santé et de la Recherche Médicale Unité 356, Institut Fédératif de Recherche 58, Université Pierre et Marie Curie, Paris, France.
J Am Soc Nephrol. 2003 Mar;14(3):545-54. doi: 10.1097/01.asn.0000050413.43662.55.
Two nonerythroid homologs of the blood group Rh proteins, RhCG and RhBG, which share homologies with specific ammonia transporters in primitive organisms and plants, could represent members of a new family of proteins involved in ammonia transport in the mammalian kidney. Consistent with this hypothesis, the expression of RhCG was recently reported at the apical pole of all connecting tubule (CNT) cells as well as in intercalated cells of collecting duct (CD). To assess the localization along the nephron of RhBG, polyclonal antibodies against the Rh type B glycoprotein were generated. In immunoblot experiments, a specific polypeptide of Mr approximately 50 kD was detected in rat kidney cortex and in outer and inner medulla membrane fractions. Immunocytochemical studies revealed RhBG expression in distal nephron segments within the cortical labyrinth, medullary rays, and outer and inner medulla. RhBG expression was restricted to the basolateral membrane of epithelial cells. The same localization was observed in rat and mouse kidney. RT-PCR analysis on microdissected rat nephron segments confirmed that RhBG mRNAs were chiefly expressed in CNT and cortical and outer medullary CD. Double immunostaining with RhCG demonstrated that RhBG and RhCG were coexpressed in the same cells, but with a basolateral and apical localization, respectively. In conclusion, RhBG and RhCG are present in a major site of ammonia secretion in the kidney, i.e., the CNT and CD, in agreement with their putative role in ammonium transport.
血型Rh蛋白的两种非红细胞同源物RhCG和RhBG,与原始生物和植物中的特定氨转运体具有同源性,可能代表参与哺乳动物肾脏氨转运的新蛋白家族成员。与这一假设一致,最近有报道称RhCG在所有连接小管(CNT)细胞的顶端以及集合管(CD)的闰细胞中表达。为了评估RhBG在肾单位中的定位,制备了针对Rh B型糖蛋白的多克隆抗体。在免疫印迹实验中,在大鼠肾皮质以及外髓和内髓膜组分中检测到一条分子量约为50 kD的特异性多肽。免疫细胞化学研究显示,RhBG在皮质迷路、髓放线以及外髓和内髓的远端肾单位节段中表达。RhBG表达局限于上皮细胞的基底外侧膜。在大鼠和小鼠肾脏中观察到相同的定位。对显微切割的大鼠肾单位节段进行的RT-PCR分析证实,RhBG mRNA主要在CNT以及皮质和外髓集合管中表达。与RhCG的双重免疫染色表明,RhBG和RhCG在同一细胞中共表达,但分别定位于基底外侧和顶端。总之,RhBG和RhCG存在于肾脏氨分泌的主要部位,即CNT和CD,这与其在铵转运中的假定作用一致。