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Articles in PresS, published online ahead of print June 4, 2002
Am J Physiol Renal Physiol, 10.1152/ajprenal.00263.2001
Submitted on August 20, 2001
Accepted on May 30, 2002
1 School of Biological Sciences, University of Manchester, Manchester, United Kingdom
2 Department of Biomedical Science, University of Sheffield, Sheffield, United Kingdom
* To whom correspondence should be addressed. E-mail: cpsmith{at}man.ac.uk.
Specialized transporter proteins that are the products of two closely related genes, UT-A (Slc14a2) and UT-B (Slc14a1), modulate the movement of urea across cell membranes. The aim of this study was to characterize the mouse variants of two major products of the UT-A gene, UT-A1 and UT-A2. Screening a mouse kidney inner medulla cDNA library yielded 4,047bp and 2,876bp cDNAs - the mouse homologues of UT-A1 and UT-A2. Northern analysis showed high levels of UT-A mRNAs in kidney medulla. UT-A transcripts were also present in testis, heart, brain and liver. Immunoblots with an antiserum raised to the 19 COOH-terminal amino acids of rat UT-A1 (L194) identified immunoreactive proteins in kidney, testis, heart, brain and liver and showed a complex pattern of differential expression. Relative to other tissues, kidney and brain had the highest levels of UT-A protein expression. In kidney sections, immunostaining with L194 revealed immunoreactive proteins in type 1 (short) and type 3 (long) thin descending limbs (tDLs) of the loop of Henle, and in the middle and terminal inner medullary collecting ducts (IMCDs). Expression in Xenopus oocytes showed that, characteristic of UT-A family members, the cDNAs encoded phloretin-inhibitable urea transporters. Acute application of protein kinase A agonists (cAMP/ forskolin/ IBMX) caused a significant increase in UT-A1 and UT-A3, but not UT-A2 mediated urea transport.
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