Immunocytochemical localization of pendrin in intercalated cell subtypes in rat and mouse kidney
Immunocytochemical localization of pendrin in intercalated cell subtypes in rat and mouse kidney
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DOI：
10.1152/ajprenal.00037.2002
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发表时间：
2002-10-01
影响因子：
4.2
通讯作者：
Nielsen, S
中科院分区：
文献类型：
--
作者：
Kim, YH;Kwon, TH;Nielsen, S
最近的研究表明，一种新的阴离子交换剂penddrin在哺乳动物集束管B型插层细胞的顶域表达。本研究的目的是：1)测定penddrin在小鼠和大鼠肾收集管和连接管中的表达和分布，并确定penddrin是否在非a -非b插层细胞中表达；2)利用免疫电镜确定penddrin在不同插层细胞群体中的细胞内定位。在大鼠和小鼠肾脏的免疫印迹中，制备了特异性识别pendrin的肽源亲和纯化抗体。免疫组织化学和共聚焦激光扫描显微镜显示，小鼠和大鼠B型插层细胞连接小管和集束管的顶端区域均存在penddrin。此外，在非a -非b插层细胞中观察到强的pendrin免疫染色。未见A型插层细胞标记。免疫电镜显示，在B型插层细胞和非a -非B型插层细胞的顶质膜和胞内囊泡中均可见pendrin；后者是通过在根尖质膜上存在H+- atp酶来鉴定的。本研究结果表明，pendrin和H+- atp酶在非a -非b插层细胞的顶质膜中均有表达，表明这些细胞既能分泌HCO3-，也能分泌质子。此外，在B型和非a -非B插层细胞的根尖质膜和根尖胞内囊泡中都存在penddrin，这表明HCO3-的分泌可能受penddrin在两个膜室之间的运输调节。
Recent studies have demonstrated that a novel anion exchanger, pendrin, is expressed in the apical domain of type B intercalated cells in the mammalian collecting duct. The purpose of this study was 1) to determine the expression and distribution of pendrin along the collecting duct and connecting tubule of mouse and rat kidney and establish whether pendrin is expressed in the non-A-non-B intercalated cells and 2) to determine the intracellular localization of pendrin in the different populations of intercalated cells by immunoelectron microscopy. A peptide-derived affinity-purified antibody was generated that specifically recognized pendrin in immunoblots of rat and mouse kidney. Immunohistochemistry and confocal laser scanning microscopy demonstrated the presence of pendrin in apical domains of all type B intercalated cells in mouse and rat connecting tubule and collecting duct. In addition, strong pendrin immunostaining was observed in non-A-non-B intercalated cells. There was no labeling of type A intercalated cells. Immunoelectron microscopy demonstrated that pendrin was located in the apical plasma membrane and intracellular vesicles of both type B intercalated cells and non-A-non-B cells; the latter was identified by the presence of H+-ATPase in the apical plasma membrane. The results of this study demonstrate that both pendrin and H+-ATPase are expressed in the apical plasma membrane of non-A-non-B intercalated cells, suggesting that these cells are capable of both HCO3- and proton secretion. Furthermore, the presence of pendrin in both the apical plasma membrane and the apical intracellular vesicles of type B and non-A-non-B intercalated cells suggests that HCO3- secretion may be regulated by trafficking of pendrin between the two membrane compartments.