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Originally published In Press as doi:10.1074/jbc.M002831200 on July 10, 2000

J. Biol. Chem., Vol. 275, Issue 38, 29754-29760, September 22, 2000
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Cell Swelling-induced Translocation of Rat Liver Na+/Taurocholate Cotransport Polypeptide Is Mediated via the Phosphoinositide 3-Kinase Signaling Pathway*

Cynthia R. L. Webster, Christopher J. Blanch, Jeffrey Phillips, and M. Sawkat AnwerDagger

From the Department of Biomedical Sciences, Tufts University School of Veterinary Medicine, North Grafton, Massachusetts 01536

Cell swelling stimulates phosphoinositide 3-kinase (PI3K) and mitogen-activated protein kinase (MAPK) in hepatocytes, and the PI3K signaling pathway is involved in cAMP-mediated translocation of sinusoidal Na+/taurocholate (TC) cotransporter (Ntcp) to the plasma membrane. We determined whether cell swelling also stimulates TC uptake and Ntcp translocation via the PI3K and/or MAPK signaling pathway. All studies were conducted in isolated rat hepatocytes. Hepatocyte swelling induced by hypotonic media resulted in: 1) time- and medium osmolarity-dependent increases in TC uptake, 2) an increase in the Vmax of Na+/TC cotransport, and 3) wortmannin-sensitive increases in TC uptake and plasma membrane Ntcp mass. Hepatocyte swelling also induced wortmannin-sensitive activation of PI3K, protein kinase B, and p70S6K. Rapamycin, an inhibitor of p70S6K, inhibited cell swelling-induced activation of p70S6K but failed to inhibit cell swelling-induced stimulation of TC uptake. Because PD98095, an inhibitor of MAPK, did not inhibit cell swelling-induced increases in TC uptake, it is unlikely that the effect of cell swelling on TC uptake is mediated via the MAPK signaling pathway. Taken together, these results indicate that 1) cell swelling stimulates TC uptake by translocating Ntcp to the plasma membrane, 2) this effect is mediated via the PI3K, but not MAPK, signaling pathway, and 3) protein kinase B, but not p70S6K, is a likely downstream effector of PI3K.

* This work was supported in part by Grant DK-33436 from the National Institutes of Health.The costs of publication of this article were defrayed in part by the payment of page charges. The article must therefore be hereby marked "advertisement" in accordance with 18 U.S.C. Section 1734 solely to indicate this fact.

Dagger To whom correspondence should be addressed: Dept. of Biomedical Sciences, Tufts University School of Veterinary Medicine, 200 Westboro Rd., North Grafton, MA 01536. Tel.: 508-839-8788; Fax: 508-839-8787; E-mail: SANWER@INFONET.TUFTS.EDU.

Copyright © 2000 by The American Society for Biochemistry and Molecular Biology, Inc.
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