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大鼠和小鼠肾脏闰细胞亚型中pendrin的免疫细胞化学定位

Immunocytochemical localization of pendrin in intercalated cell subtypes in rat and mouse kidney.

作者信息

Kim Young-Hee, Kwon Tae-Hwan, Frische Sebastian, Kim Jin, Tisher C Craig, Madsen Kirsten M, Nielsen Søren

机构信息

Department of Medicine, Hypertension and Transplantation, University of Florida, Gainesville 32610, USA.

出版信息

Am J Physiol Renal Physiol. 2002 Oct;283(4):F744-54. doi: 10.1152/ajprenal.00037.2002.

Abstract

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 HCO 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 HCO secretion may be regulated by trafficking of pendrin between the two membrane compartments.

摘要

最近的研究表明,一种新型阴离子交换蛋白——pendrin,在哺乳动物集合管B型闰细胞的顶端区域表达。本研究的目的是:1)确定pendrin在小鼠和大鼠肾脏集合管及连接小管中的表达和分布,并确定pendrin是否在非A非B型闰细胞中表达;2)通过免疫电子显微镜确定pendrin在不同类型闰细胞中的细胞内定位。制备了一种肽衍生的亲和纯化抗体,该抗体在大鼠和小鼠肾脏的免疫印迹中能特异性识别pendrin。免疫组织化学和共聚焦激光扫描显微镜显示,在小鼠和大鼠连接小管及集合管的所有B型闰细胞顶端区域存在pendrin。此外,在非A非B型闰细胞中观察到强烈的pendrin免疫染色。A型闰细胞无标记。免疫电子显微镜显示,pendrin位于B型闰细胞和非A非B型细胞的顶端质膜和细胞内囊泡中;后者通过顶端质膜中H(+) - ATP酶的存在来鉴定。本研究结果表明,pendrin和H(+) - ATP酶均在非A非B型闰细胞的顶端质膜中表达,提示这些细胞能够分泌HCO和质子。此外,pendrin在B型和非A非B型闰细胞的顶端质膜和顶端细胞内囊泡中的存在表明,HCO的分泌可能受pendrin在两个膜区室之间转运的调节。

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