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ORIGINAL RESEARCH ARTICLE

Ontogeny of Expression of Organic Anion Transporters 1 and 3 in Ovine Fetal and Neonatal Kidney

Charles E. Wood^*,1, Roderick Cousins^*, Daying Zhang^* and Maureen Keller-Wood

^* Department of Physiology and Functional Genomics, University of Florida College of Medicine; and Department of Pharmacodynamics, University of Florida College of Pharmacy, Gainesville, FL 32610

To whom requests for reprints should be addressed at ¹ Department of Physiology and Functional Genomics, P.O. Box 100274, University of Florida College of Medicine, Gainesville, FL 32610-0274 E-mail: cwood{at}phys.med.ufl.edu

Organic ions are excreted into the urine via the action of organic anion transporters (OATs). In adult kidney, both OAT1 and OAT3, both multispecific transporters, are abundant; OAT1 is a known transporter of para-aminohippurate (PAH) and OAT3 is a known transporter of sulfoconjugated estrogens. The present study was designed to test the hypotheses that the expression of both OAT1 and OAT3 are developmentally regulated and that the expression increases in late gestation. Fetal kidneys were collected at sacrifice of fetal sheep at 80, 100, 120, 130, and 145 days of gestation, as well as 1 day and 1 week after birth (n=4–5 per group). Renal tissue was separated into cortex and medulla and snap-frozen in liquid nitrogen for later extraction of mRNA. The expression levels of OAT1 and OAT3 were measured using real-time reverse transcriptase polymerase chain reaction (RT-PCR), with specific probes and primers designed in our laboratory. Cellular distribution of protein expression was identified using immunohistochemistry with commercially available antisera. The OAT1 and OAT3 mRNA in renal cortex was increased in the more mature animals. At 145 days of gestation, OAT1 mRNA abundance was increased and remained elevated postnatally. Compared with prenatal ages, OAT3 mRNA was increased postnatally. The expression of both transporters was not significantly changed as a function of development in the renal medulla. The protein expression of OAT1 and OAT3 was identified in tubular epithelium in renal cortex, although the immunoreactivity for OAT1 was greater than for OAT3. We conclude that there is a developmental pattern of expression of both OAT1 and OAT3 in ovine renal cortex, and that the pattern of expression suggests that the function of both transporters is likely to be greater starting in late gestation.

Key Words: OAT • fetal sheep • fetal kidney • qPCR