RESUMO
The identification and cloning of the urea transporter (UT) in papilla and upper pelvic epithelium of sheep kidney and the effect of a 5-week-lasting low protein diet on UT mRNAs expression in these structures are reported. Using degenerate primers we cloned by RT-PCR a 770-base pairs UT-A cDNA fragment. The deduced amino acid sequence shared 92% and 93% identity with UT-A2 protein from rabbit and rat, and from human, respectively. Quantification of UT-A mRNAs expression after LP diet was performed by quantitative RT-PCR using UT-A mutant cRNA. Compared to normal protein fed sheep, low protein diet was associated with a significant reduction of UT-A mRNA levels in pelvic epithelium (852+/-172 vs. 2024+/-260 molecules, P<0.01) and a tendency to its increase in papilla (7959+/-1741 vs. 5447+/-1040 molecules, NS). Functional studies confirmed that kidneys of low protein fed sheep increased their ability to reduce urea losses. The reduction of UT-A expression in the pelvic epithelium lining the outer medulla could be relevant for the renal conservation of urea in protein restricted sheep.