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First published online November 4, 2005
Journal of Experimental Biology 208, 4305-4315 (2005)
Published by The Company of Biologists 2005
doi: 10.1242/jeb.01879

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Transepithelial urate transport by avian renal proximal tubule epithelium in primary culture

Paul L. Dudas^1,2, Ryan M. Pelis^1,3, Eldon J. Braun³ and J. Larry Renfro^1,*

¹ Department of Physiology and Neurobiology, University of Connecticut, Storrs, Connecticut 06269, USA
² Department of Internal Medicine, Yale University School of Medicine, New Haven, CT 06520, USA
³ Department of Physiology, University of Arizona, College of Medicine, Tucson, AZ 85724, USA

View larger version (17K): [in a new window]   Fig. 1. Representative plot of unidirectional and net urate fluxes across chick PTCs as a function of time. The unidirectional fluxes include the interstitial-to-luminal secretory flux (secretory) and luminal-to-interstitial reabsorptive flux (reabsorptive; shown negative to indicate direction). Net flux is the difference between unidirectional fluxes. (A) Paired controls; (B) probenecid (1 mmol l–1) added to interstitial and luminal sides at t=0. Fluxes approached steady state at t=1.0 h.

View larger version (16K): [in a new window]   Fig. 2. Unidirectional secretory, unidirectional reabsorptive, and net urate fluxes by chick PTCs following treatment with estrone sulfate (10 µmol l–1, 500 µmol l–1, or 2500 µmol l–1). Estrone sulfate was administered to the interstitial side of the epithelium only (at t=0). Data are presented as the percentage of the paired control secretory flux and are means ± S.E.M. of four (10 µmol l–1 and 500 µmol l–1) and 10 (2500 µmol l–1) preparations. *Significantly different from paired control flux (P<0.05, paired t-test).

View larger version (18K): [in a new window]   Fig. 3. Unidirectional secretory, unidirectional reabsorptive, and net PAH fluxes by chick PTCs following treatment with urate (330 µmol l–1), urate and estrone sulfate in combination (ES + UA), or estrone sulfate (ES, 500 µmol l–1) alone. All treatments were administered to the interstitial and luminal sides of the epithelium at t=–0.5 h. Data are presented as the percentage of the paired control secretory flux after 90 min exposure and are means ± S.E.M. of three preparations. *Significantly different from paired control flux (P<0.05, paired t-test).

View larger version (15K): [in a new window]   Fig. 4. Unidirectional secretory, unidirectional reabsorptive, and net urate fluxes by chick PTCs following treatment with methotrexate (500 µmol l–1) or MK571 (20 µmol l–1). Methotrexate (at t=–0.5 h) and MK571 (at t=0) were added to the interstitial or luminal sides of the epithelium. Data are presented as the percentage of the paired control secretory flux and are means ± S.E.M. of five preparations. *Significantly different from paired control flux (P<0.05, paired t-test).

View larger version (83K): [in a new window]   Fig. 5. RT-PCR to amplify chicken renal OAT1-like, OAT3-like, MRP2-like and MRP4-like cDNA. mRNA isolated from chick PTCs was reverse transcribed, and the resulting cDNA amplified using primers to chicken OAT1, OAT3, MRP2 and MRP4 (see Table 1). The RT-PCR products were run on a 2% agarose gel and stained with Gel-Star. The cDNA template was either present (+) or absent (–). No product was obtained when the cDNA template was omitted. Molecular size (base pairs) is indicated on the left.