Ojakian G K, Romain R E, Herz R E
Am J Physiol. 1987 Sep;253(3 Pt 1):C433-43. doi: 10.1152/ajpcell.1987.253.3.C433.
Monoclonal antibodies that recognize three distinct epitopes of a 23-kDa glycoprotein (gp23) on the plasma membrane of MDCK cells were used to study cell-surface polarity. Immunofluorescence microscopy of MDCK cells obtained from the American Type Culture Collection demonstrated that gp23 was nonpolarized in approximately 50% of the cells (on both apical and basolateral membranes), whereas, in the remaining cells, gp23 had a polarized distribution (basolateral only). This heterogeneous gp23 cell-surface staining was not observed in a variety of other MDCK sublines. Instead, gp23 was found to have a nonpolarized distribution on MDCK cells that produced monolayers with low transepithelial electrical resistances (less than 220 omega X cm2) and was localized only to the basolateral membrane of MDCK cell lines capable of generating considerably higher transepithelial electrical resistances (770-2,220 omega X cm2). Immunofluorescence and immunoelectron microscopy of dog, rat, and rabbit kidney demonstrated that gp23 is a nephron segment-specific glycoprotein localized only to the distal and collecting tubules. These observations provide further evidence for the heterogeneity of the MDCK cell line. They also support a proposal that the origin of MDCK cells is the renal distal nephron.
识别MDCK细胞质膜上一种23 kDa糖蛋白(gp23)三个不同表位的单克隆抗体被用于研究细胞表面极性。对从美国模式培养物集存库获得的MDCK细胞进行免疫荧光显微镜检查显示,约50%的细胞中gp23是非极性分布的(在顶端和基底外侧膜上均有),而在其余细胞中,gp23呈极性分布(仅在基底外侧膜)。在多种其他MDCK亚系中未观察到这种异质性的gp23细胞表面染色情况。相反,在产生低跨上皮电阻(小于220 Ω×cm²)单层的MDCK细胞上,gp23呈非极性分布,而在能够产生相当高跨上皮电阻(770 - 2220 Ω×cm²)的MDCK细胞系中,gp23仅定位于基底外侧膜。对犬、大鼠和兔肾脏的免疫荧光和免疫电子显微镜检查表明,gp23是一种肾单位节段特异性糖蛋白,仅定位于远端小管和集合小管。这些观察结果为MDCK细胞系的异质性提供了进一步证据。它们还支持了MDCK细胞起源于肾远端肾单位的观点。