Expression of the mouse Na-K-2Cl cotransporter, mBSC2, in the terminal inner medullary collecting duct, the glomerular and extraglomerular mesangium, and the glomerular afferent arteriole.
Expression of the mouse Na-K-2Cl cotransporter, mBSC2, in the terminal inner medullary collecting duct, the glomerular and extraglomerular mesangium, and the glomerular afferent arteriole.
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小鼠 Na-K-2Cl 协同转运蛋白 mBSC2 在终末内髓集合管、肾小球和肾小球外系膜以及肾小球传入小动脉中的表达。
DOI：
10.1172/jci118844
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发表时间：
1996
期刊：
影响因子：
--
通讯作者：
Delpire,E
中科院分区：
文献类型：
--
作者：
Kaplan,MR;Plotkin,MD;Brown,D;Hebert,SC;Delpire,E
Na-K-Cl 共转运在肾脏厚升肢的 NaCl 重吸收中发挥重要作用，而在内髓集合管 (IMCD) 中的作用不太明确。在哺乳动物肾脏中已鉴定出两种由不同基因编码的 Na-K-Cl 协同转运蛋白：BSC1/NKCC2 定位于 Henle 的顶端厚升肢，BSC2/NKCC1 是从小鼠 IMCD 细胞系 (mIMCD-3) 中分离出来的，但其定位尚未确定。在本研究中，我们生成了针对小鼠 BSC2/NKCC1 蛋白的多克隆抗体（抗 mBSC2），以便表征和定位小鼠肾脏中的该蛋白。使用亲和纯化的抗 mBSC2 进行的蛋白质印迹分析显示，蛋白双联体为 140 和 150 kD，其在肾乳头中最丰富，但在皮质和外髓质中也可见。在免疫前血清或用特异性抗原预吸收的抗mBSC2中未观察到140-150-kD条带。免疫定位证实 mBSC2 蛋白在终末 IMCD 节段的基底外侧表面表达，并在乳头表面上皮中表达。免疫荧光还揭示了 BSC2 蛋白出人意料地存在于肾小球旁传入小动脉、可能代表肾小球外系膜的肾小球旁结构以及整个肾小球系膜。
Na-K-Cl cotransport plays an important role in the kidney in NaCl reabsorption in the thick ascending limb of Henle and a less well defined role in the inner medullary collecting duct (IMCD). Two Na-K-Cl cotransporters encoded by different genes have been identified in the mammalian kidney: BSC1/NKCC2 which localizes to the apical thick ascending limb of Henle and BSC2/NKCC1 which was isolated from a mouse IMCD cell line (mIMCD-3) but its localization has not been determined. In this study we generated a polyclonal antibody (anti-mBSC2) against the mouse BSC2/NKCC1 protein in order to characterize and localize this protein in mouse kidney. Western blot analysis with affinity-purified anti-mBSC2 showed a protein doublet of 140 and 150 kD which was most abundant in the renal papilla but also seen in cortex and outer medulla. The 140-150-kD bands were not seen with preimmune serum or with anti-mBSC2 preabsorbed with specific antigen. Immunolocalization confirmed expression of mBSC2 protein on the basolateral surface of terminal IMCD segments and demonstrated expression in the papillary surface epithelium. Immunofluorescence also revealed the unexpected presence of the BSC2 protein at the juxtaglomerular afferent arteriole, in a juxtaglomerular structure probably representing the extraglomerular mesangium, and throughout the glomerular mesangium.