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Am J Physiol Renal Physiol 291: F122-F128, 2006. First published January 31, 2006; doi:10.1152/ajprenal.00423.2005
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Urea flux across MDCK-mUT-A2 monolayers is acutely sensitive to AVP, cAMP, and [Ca2+]i

Elizabeth A. Potter, Gavin Stewart, and Craig P. Smith

Faculty of Life Sciences, The University of Manchester, Manchester, United Kingdom

Submitted 30 December 2005 ; accepted in final form 23 January 2006

In this study, we engineered a Madin-Darby canine kidney (MDCK) type I cell line to stably express the mouse urea transporter UT-A2. Monolayers of MDCK-mUT-A2 cells had a basal phloretin-inhibitable urea permeability of 8.4 x 10–6 ± 0.3 cm/s. Treatment of MDCK-mUT-A2 monolayers with AVP led to a rapid dose-dependent increase in trans-monolayer phloretin-inhibitable urea flux. The temporal pattern of response was markedly different from that observed for MDCK cells expressing rat UT-A1. Exposure of MDCK-mUT-A2 cells to either 10 µM forskolin or 250 µM 8-bromo cAMP also increased urea flux rate. Inclusion of the PKA inhibitor H89 (10 µM) had no effect on the forskolin-stimulated increase in urea flux across MDCK-mUT-A2 monolayers. Treatment with either 10 µM CPA or 1 mM ATP also caused an increase in UT-A2-mediated urea flux, although these responses where transient compared with those induced by AVP or elevated cAMP. Taken together, these results show for the first time that UT-A2 is acutely sensitive to AVP, cAMP, or increased intracellular calcium.

arginine vasopressin; intracellular calcium; urine concentration; urea transporter


Address for reprint requests and other correspondence: C. P. Smith, Faculty of Life Sciences, Univ. of Manchester, G.38, Stopford Bldg., Oxford Road, Manchester, M13 9PT, UK (e-mail: craig.smith{at}manchester.ac.uk)




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