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1 Renal Division, Department of Medicine, Emory University School of Medicine, Atlanta, Georgia, United States
2 Department of Physiology and Biophysics, Weill Medical College of Cornell University, New York, New York, United States
3 Wellcome Trust Genome Campus, The Wellcome Trust Sanger Institute, Cambridge, United Kingdom
4 Renal Division, Department of Medicine and Department of Physiology, Emory University School of Medicine, Atlanta, Georgia, United States
* To whom correspondence should be addressed. E-mail: vpech{at}emory.edu.
Pendrin (Slc26a4) localizes to type B and non-A, non-B intercalated cells in the distal convoluted tubule, the connecting tubule and the cortical collecting duct (CCD), where it mediates apical Cl-/HCO3- exchange. The purpose of this study was to determine if angiotensin II increases transepithelial net chloride transport, JCl, in mouse CCD through a pendrin-dependent mechanism. JCl and transepithelial voltage, VT, were measured in CCDs perfused in vitro from wild type and Slc26a4 null mice ingesting a NaCl-replete diet or a NaCl-replete diet and furosemide. In CCDs from wild type mice ingesting a NaCl-replete diet, VT and JCl were not different from zero either in the presence or absence of angiotensin II (10-8 M) in the bath. Thus, further experiments employed mice treated with the high NaCl diet and furosemide to upregulate pendrin expression in kidney. CCDs from furosemide-treated wild type mice had a lumen negative VT and absorbed Cl-. With angiotensin II in the bath, Cl- absorption doubled although VT did not become more lumen-negative. In contrast, in CCDs from furosemide-treated Slc26a4 null mice, Cl- secretion and a VT of ~0 were observed, neither of which changed with angiotensin II application. Inhibiting ENaC with benzamil abolished VT although JCl fell only ~50%. Thus substantial Cl- absorption is observed in the absence of an electromotive force. Attenuating apical anion exchange with the peritubular application of the H+-ATPase inhibitor, bafilomycin, abolished benzamil-insensitive Cl- absorption. In conclusion, angiotensin II increases transcellular Cl- absorption in the CCD through a pendrin- and H+-ATPase-dependent process.
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