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Originally published In Press as doi:10.1074/jbc.M203803200 on May 13, 2002

J. Biol. Chem., Vol. 277, Issue 30, 26934-26943, July 26, 2002
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Impaired Organic Anion Transport in Kidney and Choroid Plexus of Organic Anion Transporter 3 (Oat3 (Slc22a8)) Knockout Mice*

Douglas H. SweetDagger , David S. Miller§, John B. Pritchard§, Yuko Fujiwara, David R. Beier||, and Sanjay K. NigamDagger **

From the Dagger  Departments of Pediatrics, Medicine (Division of Nephrology/Hypertension), and Cellular and Molecular Medicine, University of California, San Diego, La Jolla, California 92093, the § Laboratory of Pharmacology and Chemistry, NIEHS, National Institutes of Health, Research Triangle Park, North Carolina 27709, the  Division of Hematology/Oncology, Children's Hospital, Howard Hughes Medical Institute, Harvard Medical School, Boston, Massachusetts 02115, and the || Genetics Division, Brigham and Women's Hospital, Harvard Medical School, Boston, Massachusetts 02115

To begin to develop in vivo model systems for the assessment of the contributions of specific organic anion transporter (OAT) family members to detoxification, development, and disease, we carried out a targeted disruption of the murine organic anion transporter 3 (Oat3) gene. Surviving Oat3-/- animals appear healthy, are fertile, and do not exhibit any gross morphological tissue abnormalities. No Oat3 mRNA expression was detected in kidney, liver, or choroid plexus (CP) of Oat3-/- mice. A distinct phenotype manifested by a substantial loss of organic anion transport capacity in kidney and CP was identified. Uptake sensitive to inhibition by bromosulfophthalein or probenecid was observed for taurocholate, estrone sulfate, and para-aminohippurate in renal slices from wild-type mice, whereas in Oat3-/- animals transport of these substances was greatly reduced. No discernable differences in uptake were observed between hepatic slices from wild-type and Oat3-/- littermates, suggesting Oat3 does not play a major role in hepatic organic anion uptake. Cellular accumulation of fluorescein was reduced by ~75% in CP from Oat3-/- mice. However, capillary accumulation of fluorescein-methotrexate was unchanged, indicating the effects of Oat3 loss are restricted to the entry step and that Oat3 is localized to the apical membrane of CP. These data indicate a key role for Oat3 in systemic detoxification and in control of the organic anion distribution in cerebrospinal fluid.

* This work was supported by National Institutes of Health CHHD Grant R01-HD40011 (to S. K. N.) and a grant from the March of Dimes Foundation (to D. R. B.).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.

** To whom correspondence should be addressed: Depts. of Pediatrics, Medicine, and Cellular and Molecular Medicine, Division of Nephrology/Hypertension, University of California, San Diego, 9500 Gilman Dr. 0693, La Jolla, CA 92093. Tel.: 858-822-3482; Fax: 858-822-3483.

Copyright © 2002 by The American Society for Biochemistry and Molecular Biology, Inc.
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