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Am J Physiol Renal Physiol 287: F628-F638, 2004. First published May 18, 2004; doi:10.1152/ajprenal.00363.2003
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Basolateral ammonium transport by the mouse inner medullary collecting duct cell (mIMCD-3)

Mary E. Handlogten,1 Seong-Pyo Hong,1 Connie M. Westhoff,2,3 and I. David Weiner4,1

4Renal Section, North Florida/South Georgia Veterans Health System, 1Division of Nephrology, Hypertension, and Transplantation, University of Florida College of Medicine, Gainesville, Florida 32610-0224; 2Pathology and Laboratory Medicine, University of Pennsylvania, 3The American Red Cross Philadelphia, Pennsylvania 19104

Submitted 15 October 2003 ; accepted in final form 17 May 2004

The renal collecting duct is the primary site for the ammonia secretion necessary for acid-base homeostasis. Recent studies have identified the presence of putative ammonia transporters in the collecting duct, but whether the collecting duct has transporter-mediated ammonia transport is unknown. The purpose of this study was to examine basolateral ammonia transport in the mouse collecting duct cell (mIMCD-3). To examine mIMCD-3 basolateral ammonia transport, we used cells grown to confluence on permeable support membranes and quantified basolateral uptake of the radiolabeled ammonia analog [14C]methylammonia ([14C]MA). mIMCD-3 cell basolateral MA transport exhibited both diffusive and transporter-mediated components. Transporter-mediated uptake exhibited a Km for MA of 4.6 ± 0.2 mM, exceeded diffusive uptake at MA concentrations below 7.0 ± 1.8 mM, and was competitively inhibited by ammonia with a Ki of 2.1 ± 0.6 mM. Transporter-mediated uptake was not altered by inhibitors of Na+-K+-ATPase, Na+-K+-2Cl cotransporter, K+ channels or KCC proteins, by excess potassium, by extracellular sodium or potassium removal or by varying membrane potential, suggesting the presence of a novel, electroneutral ammonia-MA transport mechanism. Increasing the outwardly directed transmembrane H+ gradient increased transport activity by increasing Vmax. Finally, mIMCD-3 cells express mRNA and protein for the putative ammonia transporter Rh B-glycoprotein (RhBG), and they exhibit basolateral RhBG immunoreactivity. We conclude that mIMCD-3 cells express a basolateral electroneutral NH4+/H+ exchange activity that may be mediated by RhBG.

ammonia; Rh B-glycoprotein



Address for reprint requests and other correspondence: I. D. Weiner, Univ. of Florida College of Medicine, PO Box 100224, Gainesville, FL 32610-0224 (E-mail: weineid{at}ufl.edu)




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