受体Megalin和Cubilin在Opossum肾小管细胞的表达及定位

Localization of Megalin and Cubilin in Proximal Tubule Cells of Opossum Kidney

目的 检测肾小管细胞膜大分子糖蛋白受体Megalin和Cubilin在Opossum肾小管 (OK)细胞的表达及亚细胞分布。方法 采用Western印迹法、免疫荧光双标记法及免疫细胞化学双标记法。结果 受体Megalin及Cubilin在分离的OK细胞膜上有表达 ,荧光双标记显示两种受体十分相似地分布在细胞周边部 ,电镜下 ,标记金颗粒的两种受体相近地分布在细胞顶部胞质的细胞内吞系统。结论 两种受体在OK细胞上的分布特征与两种受体在体内肾近端小管小皮细胞的分布相似 ,提示OK细胞可成为研究两种受体介导可能的、未知的配体细胞内吞功能的有效细胞株 。

Objective The present study aims at detecting the colocalization of two giant glycoprotein receptors, Megalin and Cubilin, in cultured proximal tubule cells of opossum kidney (OK). Method The experiments were carried out by using Western blotting, immunofluoresence and immunocytochemistry techniques. Results The expression of both Megalin and Cubilin on purified membranes of OK cells was analyzed by Western blotting using polyclonal anti Megalin and Cubilin antibodies. The cellular and subcellular colocalization was demonstrated by a double labeling immunofluoresence and immunocytochemistry techniques, respectively. Both receptors were similarly distributed at cytoplasmic apical endocytic apparatus. Conclusion The present study demonstrates membrane associated receptor, Megalin and Cubilin, in OK cells and established their subcellular colocalization, thereby identifying a suitable model system to study the receptor receptor interaction to assist to uptake of their ligands.