Identification of distinct subpopulations of intercalated cells in the mouse collecting duct.
Identification of distinct subpopulations of intercalated cells in the mouse collecting duct.
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DOI：
10.1681/asn.v72260
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发表时间：
1996-02
期刊：
影响因子：
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通讯作者：
P. Teng‐umnuay;J. Verlander;Weiping Yuan;C. Tisher;Kirsten M. Madsen
中科院分区：
文献类型：
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作者：
P. Teng‐umnuay;J. Verlander;Weiping Yuan;C. Tisher;Kirsten M. Madsen
在大鼠和兔的集合管中，已经描述了结构和功能上不同的间质细胞群。然而，对小鼠肾脏中的这些细胞知之甚少。本文研究了小鼠不同类型间充质细胞的超微结构和免疫学特征。用1%戊二醛或多聚甲醛-苦味酸固定剂活体保存正常雌性小鼠C57BL/6和IBR的肾脏，分别进行形态观察和光镜、电子显微镜免疫组织化学染色。用抗碳酸酐酶II、H+-ATPase和Band 3蛋白的抗体对IS切片进行亲和素-生物素-辣根过氧化物酶程序。用H+-ATPase和Band 3蛋白抗体对Lowicryl切片进行免疫金细胞化学染色。H+-ATPase和Band 3蛋白的共定位采用免疫金银增强双标记技术。H+-ATPase和碳酸氢酶II染色阳性的间质细胞占连接小管、皮质集合管和外髓集合管细胞总数的35%~40%。带3染色阳性的A型细胞分别占CNT、Ccd和OMCD细胞总数的16%、24%和33%。电子显微镜和免疫金细胞化学显示有三种不同的间质细胞群。A型细胞的顶端H+-ATPase和基底侧带3免疫反应阳性，在根尖表面下有明显的根尖微突起和特征性的管泡状结构，细胞质表面均覆盖有螺柱。在最初的集合管中，B型插入细胞最常见，但在CNT和早期的CD中也有表达。B型细胞顶端表面光滑，顶端质膜下无细胞器的灰色带，细胞内有小的无螺柱的胞质小泡。第三种类型的插入细胞仅见于CNT和最初的集合管，具有顶端和细胞质的H+-ATPase，但没有基侧带3蛋白。这种类型的细胞很大，有许多线粒体，细胞内到处都有包裹着螺柱的小泡。它类似于之前在大鼠身上描述的第三种类型的间质细胞。结论：在小鼠肾脏中存在三种不同的形态和免疫学类型的嵌合细胞。
Structurally and functionally distinct populations of intercalated cells have been described in the collecting duct of both rat and rabbit. However, little is known about these cells in the mouse kidney. The study presented here examines ultrastructural and immunological characteristics of different types of intercalated cells in the mouse. Kidneys of two strains of normal female mice, C57BL/6 and IBR, were preserved by in vivo perfusion with 1% glutaraldehyde or paraformaldehyde-picric acid fixatives and processed for morphological evaluation or light and electron microscopic immunohistochemistry, respectively. The avidin-biotin-horseradish peroxidase procedure was performed on was sections using antibodies against carbonic anhydrase II, H+ -ATPase and Band 3 protein. Immunogold cytochemistry was performed on Lowicryl sections using antibodies to H+ -ATPase and Band 3 protein. Colocalization of H+ -ATPase and Band 3 protein was performed by double labeling using an immunogold technique with silver enhancement. Intercalated cells identified by positive staining for H+ -ATPase and carbonic anhydrase II constituted 35% to 40% of all cells in the connecting tubule (CNT), cortical collecting duct (CCD), and outer medullary collecting duct (OMCD). Type A intercalated cells identified by positive Band 3 staining constituted 16%, 24%, and 33% of the total cell population in the CNT, CCD, and OMCD, respectively. Electron microscopy and immunogold cytochemistry demonstrated three distinct populations of intercalated cells. Type A intercalated cells with apical H+ -ATPase and basolateral Band 3 immunoreactivity were present in all segments examined, and had prominent apical microprojections and characteristic tubulovesicular structures beneath the apical surface, both coated with studs on the cytoplasmic face. Type B intercalated cells with basolateral and cytoplasmic H+-ATPase and no Band 3 immunoreactivity were most frequently observed in the initial collecting tubule, but were present also in the CNT and early CCD. Type B intercalated cells had a fairly smooth apical surface, a gray zone free of organelles beneath the apical plasma membrane, and small cytoplasmic vesicles without studs throughout the cell. A third type of intercalated cell with apical and cytoplasmic H+-ATPase, but no basolateral Band 3 protein, was observed exclusively in the CNT and the initial collecting tubule. This type of cell was large, with numerous mitochondria, and vesicles coated with studs were present throughout the cell. It resembled a third type of intercalated cell described previously in the rat. It is concluded that three morphologically and immunologically distinct types of intercalated cells are present in the mouse kidney.