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Originally published In Press as doi:10.1074/jbc.M108472200 on September 11, 2001

J. Biol. Chem., Vol. 276, Issue 45, 41611-41619, November 9, 2001
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Ventricular Choline Transport
A ROLE FOR ORGANIC CATION TRANSPORTER 2 EXPRESSED IN CHOROID PLEXUS*

Douglas H. SweetDagger , David S. Miller, and John B. Pritchard

From the Laboratory of Pharmacology and Chemistry, NIEHS, National Institutes of Health, Research Triangle Park, North Carolina 27709

To determine whether organic cation transporter (OCT) family members might mediate choline transport in choroid plexus (CP), the handling of choline by cloned transporters and by intact CP isolated from the adult rat was investigated. Expression of OCT1 and OCT2 in Xenopus oocytes increased hemicholinium-3-sensitive choline uptake. In contrast, OCT3 did not mediate choline transport. Estimated Km values for choline in rOCT1-, rOCT2-, and hOCT2-expressing oocytes were 346 ± 50, 441 ± 67, and 102 ± 80 µM, respectively. Membrane potential was the major driving force for choline uptake in rat and human OCT2-expressing oocytes and in intact CP in vitro. Lowering of medium pH (6 versus 7.4) was equally effective at inhibiting choline uptake in CP, suggesting that there might be a non-OCT component of choline uptake that is responsive to an H+ gradient. However, choline efflux from CP was not stimulated by a trans-applied H+ gradient. Choline uptake by CP was Na+-independent with an estimated Km of 183 µM. Reverse transcriptase-polymerase chain reaction detected OCT2 and OCT3, but not OCT1, mRNA expression in CP. Transfection of intact CP with a rOCT2/green fluorescent protein fusion construct resulted in strong apical membrane fluorescence with no detectable signal in the basal and lateral plasma membranes. These data indicate that OCT2 mediates choline transport across the ventricular membrane of CP.

* 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 Medicine 0693, University of California San Diego, 9500 Gilman Dr., La Jolla, CA 92093-0693. Tel.: 858-822-3382; Fax: 858-822-3483; E-mail: dosweet@ucsd.edu.

Copyright © 2001 by The American Society for Biochemistry and Molecular Biology, Inc.
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