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

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.