The intracellular trafficking pathway of
albumin in podocytes remains controversial. We therefore analysed
albumin endocytosis through caveolae, subsequent transcytosis, and exocytosis. In Western blot and immunofluorescence analysis in vitro,
methyl-beta-cyclodextrin (MBCD) treatment significantly decreased the expression of
caveolin-1 and
albumin in cultured human podocytes after incubation with
albumin; additionally, MBCD interfered with
albumin endocytosis through caveolae in the experiment using Transwell plates. In the immunofluorescence analysis,
albumin was incubated with cultured human podocytes, and colocalisation analysis with organelles and cytoskeletons in the podocytes showed that
albumin particles colocalised with
caveolin-1 and
Fc-receptor but not
clathrin in endocytosis, colocalised with actin cytoskeleton but not microtubules in transcytosis, and colocalised with early endosomes and lysosomes but not
proteasome, endoplasmic reticulum, or Golgi apparatus. In the electron microscopic analysis of podocytes in
nephrotic syndrome model mice,
gold-labelled
albumin was shown as endocytosis, transcytosis, and exocytosis with caveolae. These results indicate the intracellular trafficking of
albumin through podocytes.
Albumin enters through caveolae with the
Fc-receptor, moves along actin, and reaches the early endosome, where some of them are sorted for lysosomal degradation, and others are directly transported outside the cells through exocytosis. This intracellular pathway may be a new aetiological hypothesis for
albuminuria.