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Articles in PresS, published online ahead of print August 13, 2002
Am J Physiol Renal Physiol, 10.1152/ajprenal.00055.2002
Submitted on February 7, 2002
Accepted on August 7, 2002
1 Division of Nephrology, Department of Medicine, University of Cinnati, Cincinnati, OH, USA
2 Veterans Affairs Medical Center, Cincinnati, OH, USA; Division of Nephrology, Department of Medicine, University of Cinnati, Cincinnati, OH, USA
3 Division of Nephrology, Department of Medicine, University of Cinnati, Cincinnati, OH, USA; Veterans Affairs Medical Center, Cincinnati, OH, USA
* To whom correspondence should be addressed. E-mail: manoocher.soleimani{at}uc.edu.
The purpose of the present studies was to examine the renal distribution and functional properties of NBC4, the latest Na+:HCO3- cotransporter isoform to be identified. Zonal distribution studies in rat kidney by Northern hybridization and RT-PCR demonstrated that NBC4 is highly abundant in the outer medulla and cortex but is low in the inner medulla. Nephron segment distribution studies indicated that NBC4 in predominantly expressed in medullary and cortical thick ascending limb of Henle. Using specific primers based on published sequence (GenBank accession number AF 207661), a full length NBC4 variant was cloned from human liver and examined. The sequence of this variant (called NBC4e) is shorter by 86 aa vs. the published sequence. Xenopus oocytes injected with the full length NBC4e cRNA were compared to NBC1-expressing oocytes. Whereas exposure of NBC1-expressing oocytes to CO2/HCO3- resulted in immediate hyperpolarization, the NBC4-expressing oocytes did not show any alteration in membrane potential. NBC activity in oocytes, assayed as the Na-dependent, HCO3-mediated pHi recovery from acidosis, indicated that NBC4 is a DIDS-inhibitable, Na+:HCO3- cotransporter. We propose that NBC4 is expressed in the thick ascending limb of Henle and mediates cellular HCO3- uptake in this nephron segment.
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