title: Cadmium inhibits vacuolar H(+)-ATPase and endocytosis in rat kidney cortex.
abstract: The mechanism of cadmium (Cd)-induced damage in the mammalian proximal tubule that is manifested by defects in reabsorption of various compounds, is poorly understood. A vacuolar H(+)-ATPase (V-ATPase) in proximal tubule (PT) brush border and intracellular vesicles may be affected by Cd, and this may influence intracellular vesicle trafficking and reabsorption of the filtered proteins. We studied the effects of Cd on V-ATPase and endocytosis in rat renal PT in vivo and on acidification mechanisms in isolated renal cortical organelles in vitro. The V-ATPase activity in brush border membrane (BBM) from Cd-intoxicated rats was 40% lower compared to that in control animals. Immunofluorescence studies in cortical tissue sections and Western blot studies in BBM from Cd-treated rats showed a strongly decreased abundance of the 31 kDa and 70 kDa V-ATPase subunits. Functional studies in vivo showed a dramatically diminished endocytosis of fluorescein-labeled dextran in PT cells from Cd-treated animals, whereas morphological studies revealed a loss of endocytic invaginations and subapical vesicles in the same cells. In studies in vitro, Cd inhibited V-ATPase activity in a concentration- and time-dependent manner in both BBM and endocytic vesicles, whereas in endocytic vesicles, Cd inhibited ATP-driven intravesicular acidification and accelerated the dissipation of transmembrane pH gradients. We conclude that Cd may impair acidification in cell organelles by (a) causing a loss of V-ATPase protein in their limiting membranes, (b) inhibiting the intrinsic V-ATPase activity, and (c) dissipating the transmembrane pH gradient. This may inhibit endocytosis of filtered proteins and impair vesicle-mediated recycling of some membrane transporters, thus contributing to the loss of reabsorptive capacity of the PT.
authors.affiliation: Institute for Medical Research and Occupational Health, Zagreb, Croatia.
authors.forename: C M
authors.initials: CM
authors.lastname: Herak-Kramberger
authors.forename: D
authors.initials: D
authors.lastname: Brown
authors.forename: I
authors.initials: I
authors.lastname: Sabolić
mesh_terms: D000136:Acid-Base Equilibrium; D000818:Animals; D002104:Cadmium / Q000494:pharmacology*; D003911:Dextrans; D004705:Endocytosis / Q000187:drug effects*; D016650:Fluorescein-5-isothiocyanate / Q000031:analogs & derivatives; D007668:Kidney / Q000187:drug effects / Q000503:physiopathology; D007672:Kidney Cortex / Q000201:enzymology* / Q000502:physiology* / Q000648:ultrastructure; D008854:Microscopy, Electron; D006180:Proton-Translocating ATPases / Q000037:antagonists & inhibitors*; D051381:Rats; D017208:Rats, Wistar; D014617:Vacuoles / Q000201:enzymology*
chemical_list: D003911:Dextrans; C015219:fluorescein isothiocyanate dextran; D002104:Cadmium; D006180:Proton-Translocating ATPases; D016650:Fluorescein-5-isothiocyanate