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Am J Physiol Cell Physiol 253: C721-C730, 1987;
0363-6143/87 $5.00
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AJP - Cell Physiology, Vol 253, Issue 5 C721-C730, Copyright © 1987 by American Physiological Society


ARTICLES

(Na + K + 2Cl) cotransport in cultured embryonic chick heart cells

S. Liu, R. Jacob, D. Piwnica-Worms and M. Lieberman
Department of Physiology, Duke University Medical Center, Durham, North Carolina 27710.

The coupled movements of Na, K, and Cl were studied in cultured chick embryonic heart cells using ion-selective microelectrodes. Movements of K and Cl in response to changes in extracellular [K] ([K]o) showed a furosemide-sensitive coupled process. The movement of Na was then studied. Lowering extracellular [Na] ([Na]o) to 27 mM caused a decrease in intracellular Cl activity (aicl). Upon restoring [Na]o to 143 mM, Cl was taken up against its electrochemical gradient (delta mu Cl). In Cl-free solution, cells lost Na against delta mu Na and simultaneously lost Cl. Upon restoring extracellular [Cl] ([Cl]o), Cl was taken up against delta mu Cl; this was accompanied by an uptake of Na. The Cl uptake was 4-acetamido-4'-isothiocyanostilbene-2,2'-disulfonic acid (SITS)-insensitive (0.1 mM) but inhibited by removing Nao. Both Cl and Na uptakes were potentiated by raising [K]o from 5.4 to 15 mM, and Na uptake was diminished by lowering [K]o to 1 mM. In all experiments, Cl and Na movements were furosemide (0.3 mM) or bumetanide-sensitive (0.1 mM). Removal of Nao, with resultant depletion of intracellular [Na] ([Na]i), blocked the furosemide or bumetanide-sensitive Cl loss or uptake upon exposure to zero or 133 mM [K]o + SITS (0.1 mM), respectively. These results suggest that cultured heart cells possess an electroneutral (Na + K + 2Cl) cotransport.


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