AJP - Gastrointestinal and Liver Physiology, Vol 267, Issue 6 1004-G1011, Copyright © 1994 by American Physiological Society

ARTICLES

Na-K-Cl cotransport in villus and crypt cells from rat duodenum

C. M. McNicholas, C. D. Brown and L. A. Turnberg
Epithelial Membrane Research Centre, Hope Hospital, Salford, United Kingdom.

The aim of the present study was to examine the possibility that Cl- uptake into both villus and crypt epithelial cells of rat duodenum occurs via an electroneutral Na-K-Cl coupled-transport mechanism. Sheets of villus cells and whole crypts were isolated using a Ca2+ chelation technique combined with continuous vibration at low temperatures. Structurally intact, viable epithelia from defined regions along the villus-crypt axis were produced. Uptake of 86Rb+ (as a proxy for K+) into both villus and crypt cells appeared to depend on a coupled process, as evidenced by the inhibition of 86Rb+ uptake by bumetanide and by the removal of either Na+ or Cl- from the bathing media. We report an improved method of isolation of viable enterocytes from defined regions along the villus-crypt axis. We demonstrate the presence of Na-K-Cl cotransport in both villus and crypt duodenal enterocytes.

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