Role of CFTR and ClC-5 in modulating vacuolar H-ATPase activity in kidney proximal tubule

Department of Physiology and Biophysics, University of São Paulo, São Paulo, Brazil.
Cellular Physiology and Biochemistry (Impact Factor: 2.88). 10/2010; 26(4-5):563-76. DOI: 10.1159/000322324
Source: PubMed


It has been widely accepted that chloride ions moving along chloride channels act to dissipate the electrical gradient established by the electrogenic transport of H(+) ions performed by H(+)-ATPase into subcellular vesicles. Largely known in intracellular compartments, this mechanism is also important at the plasma membrane of cells from various tissues, including kidney. The present work was performed to study the modulation of plasma membrane H(+)-ATPase by chloride channels, in particular, CFTR and ClC-5 in kidney proximal tubule.
Using in vivo stationary microperfusion, it was observed that ATPase-mediated HCO(3)(-) reabsorption was significantly reduced in the presence of the Cl(-) channels inhibitor NPPB. This effect was confirmed in vitro by measuring the cell pH recovery rates after a NH(4)Cl pulse in immortalized rat renal proximal tubule cells, IRPTC. In these cells, even after abolishing the membrane potential with valinomycin, ATPase activity was seen to be still dependent on Cl(-). siRNA-mediated CFTR channels and ClC-5 chloride-proton exchanger knockdown significantly reduced H(+)-ATPase activity and V-ATPase B2 subunit expression.
These results indicate a role of chloride in modulating plasma membrane H(+)-ATPase activity in proximal tubule and suggest that both CFTR and ClC-5 modulate ATPase activity.

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    • "Carcinoma renal murine (Renca) [16] and immortalized rat proximal tubule cells (IRPTC) [17] were kindly provided by Dr. Maria Helena Bellini (Institute of Energy and Nuclear Research, IPEN, São Paulo, Brazil) and Dr. Maria Oliveira de Souza (Department of Physiology and Biophysics, Institute of Biomedical Sciences, University of São Paulo, São Paulo, Brazil), respectively. Human umbilical vein endothelial cells (HUVEC -CRL 1730) were obtained from the American Type Culture Collection (Mannasa, VA, USA). "
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