Angiotensin II stimulates trafficking of NHE3, NaPi2, and associated proteins into the proximal tubule microvilli

1 Department of Cell and Neurobiology, University of Southern California Keck School of Medicine, Los Angeles, California; and ; 2 The Water and Salt Research Center, Department of Cell Biology, Institute of Anatomy, University of Aarhus, Aarhus, Denmark Submitted August 12, 2009 ; accepted in final form October 22, 2009 Angiotensin II (ANG II) stimulates proximal tubule (PT) sodium and water reabsorption. We showed that treating rats acutely with the angiotensin-converting enzyme inhibitor captopril decreases PT salt and water reabsorption and provokes rapid redistribution of the Na + /H + exchanger isoform 3 (NHE3), Na + /Pi cotransporter 2 (NaPi2), and associated proteins out of the microvilli. The aim of the present study was to determine whether acute ANG II infusion increases the abundance of PT NHE3, NaPi2, and associated proteins in the microvilli available for reabsorbing NaCl. Male Sprague-Dawley rats were infused with a dose of captopril (12 µg/min for 20 min) that increased PT flow rate 20% with no change in blood pressure (BP) or glomerular filtration rate (GFR). When ANG II (20 ng·kg –1 ·min –1 for 20 min) was added to the captopril infusate, PT volume flow rate returned to baseline without changing BP or GFR. After captopril, NHE3 was localized to the base of the microvilli and NaPi2 to subapical cytoplasmic vesicles; after 20 min ANG II, both NHE3 and NaPi2 redistributed into the microvilli, assayed by confocal microscopy and density gradient fractionation. Additional PT proteins that redistributed into low-density microvilli-enriched membranes in response to ANG II included myosin VI, DPPIV, NHERF-1, ezrin, megalin, vacuolar H + -ATPase, aminopeptidase N, and clathrin. In summary, in response to 20 min ANG II in the absence of a change in BP or GFR, multiple proteins traffic into the PT brush-border microvilli where they likely contribute to the rapid increase in PT salt and water reabsorption. hypertension; captopril Address for reprint requests and other correspondence: A. A. McDonough, Dept. of Cell and Neurobiology, Univ. of Southern California Keck School of Medicine, 1333 San Pablo St., BMT 403, Los Angeles, CA 90089 (e-mail: mcdonoug{at}usc.edu ).