Expression of urea transporter (UT-A) mRNA in papilla and pelvic epithelium of kidney in normal and low protein fed sheep

Abstract

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.