Cubilin- and megalin-mediated uptake of albumin in cultured proximal tubule cells of opossum kidney

Citation
Xy. Zhai et al., Cubilin- and megalin-mediated uptake of albumin in cultured proximal tubule cells of opossum kidney, KIDNEY INT, 58(4), 2000, pp. 1523-1533
Citations number
42
Language
INGLESE
art.tipo
Article
Categorie Soggetti
Urology & Nephrology","da verificare
Journal title
KIDNEY INTERNATIONAL
ISSN journal
0085-2538 → ACNP
Volume
58
Issue
4
Year of publication
2000
Pages
1523 - 1533
Database
ISI
SICI code
0085-2538(200010)58:4<1523:CAMUOA>2.0.ZU;2-B
Abstract
Background. Reabsorption of albumin from the glomerular filtrate occurs via receptor-mediated endocytosis in the proximal tubule. This process is init iated by binding of albumin in apical clathrin-coated pits, followed by end ocytosis and degradation in lysosomes. Although binding sites have been cha racterized by kinetic studies, the receptors responsible for the binding of albumin have not been fully identified. Two giant glycoproteins, cubilin a nd megalin, constitute important endocytic receptors localized to the kidne y proximal tubule. Methods. In the present study, we examined the colocalization of cubilin an d megalin in the endocytic pathway and the relationship between the uptake of albumin and the expression of cubilin and megalin in opossum kidney (OK) proximal tubule cells by immunocytochemistry and immunoblotting. Results. OK cells expressed both cubilin and megalin. The light microscope labeling patterns for cubilin and megalin were almost identical and were ma inly located at the surface area of the cells. Cubilin and megalin were als o shown to colocalize on cell surface microvilli, in coated pits, and in en docytic compartments at the electron microscope level. Endocytosed bovine s erum albumin (BSA) was identified exclusively in cells expressing megalin a nd cubilin. Uptake of BSA-FITC was saturable and inhibited by receptor-asso ciated protein (RAP) and by intrinsic factor-vitamin B-12 complex (IF-B-12) at high concentrations. Significant inhibition was also observed by specif ic antibodies to cubilin, and megalin and cubilin antisense oligonucleotide s likewise significantly reduced albumin uptake. Egg albumin did not affect the uptake of BSA. Conclusion. The present observations suggest that the two receptors cubilin and megalin are both involved in the endocytic uptake of albumin in renal proximal tubule cells.