Amiloride transport in rabbit renal brush-border membrane vesicles

S. H. Wright, T. M. Wunz

Research output: Contribution to journalArticle

17 Scopus citations

Abstract

Rabbit renal brush-border membrane vesicles (BBMV) were used to study amiloride transport across the luminal membrane of proximal tubular cells. An outwardly directed H+ gradient (pH(i) 6.0; pH(o) 7.5) stimulated 8 μM [14C]-amiloride uptake into BBMV and supported a transient 'active' accumulation of substrate consistent with the presence of an amiloride-H+ exchange process. Uptake was inhibited, in the presence or absence of a pH gradient, by 1 mM unlabeled amiloride or 20 mM tetraethylammonium (TEA). Amiloride transport was not directly affected by the presence of 100 mM Na+ in the extravesicular medium, suggesting that Na-H exchange did not mediate amiloride flux. Amiloride transport was a saturable process with a maximal flux (under pH gradient conditions) of 3 nmol·mg-1·min-1 and an apparent K(t) of 8 μM. TEA acted as a competitive inhibitor of this process with an apparent K(i) of ~80 μM, similar to the K(t) of TEA transport via the TEA-H+ exchanger. Likewise, amiloride acted as a competitive inhibitor of TEA uptake with an apparent K(i) of ~11 μM. Preloading BBMV with 1-2 mM TEA stimulated the rate of amiloride uptake and supported a transient active accumulation of amiloride. We conclude that amiloride and TEA are transported by a common pathway in BBMV, which involves a carrier-mediated exchange with H+ and which may play a role in the tubular secretion of these compounds.

Original languageEnglish (US)
Pages (from-to)F462-F468
JournalAmerican Journal of Physiology - Renal Fluid and Electrolyte Physiology
Volume256
Issue number3 (25/3)
DOIs
StatePublished - Jan 1 1989

ASJC Scopus subject areas

  • Physiology

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