Water transport mediated by murine urea transporters: implications for urine concentration mechanisms

• Main Authors: J. Kabutomori, N. Pina-Lopes, R. Musa-Aziz
• Format: Article
• Language: English
• Published: The Company of Biologists 2020-08-01
• Series: Biology Open
• Subjects: urea transporter; water transport; concentrated urine; lithobates oocytes; membrane permeability; renal physiology
• Online Access: http://bio.biologists.org/content/9/8/bio051805

Urea transporters (UTs) facilitate urea diffusion across cell membranes and play an important role in the urinary concentration mechanisms in the kidney. Herein, we injected cRNAs encoding for c-Myc-tagged murine UT-B, UT-A2 or UT-A3 (versus water-injected control) in Lithobates oocytes and evaluated oocyte surface protein expression with biotinylation and immunoblotting, urea uptake using [14C] counts and water permeability (Pf) by video microscopy. Immunoblots of UT-injected oocyte membranes revealed bands with a molecular weight consistent with that of a UT monomer (34 kDa), and UT-injected oocytes displayed significantly increased and phloretin-sensitive urea uptake and Pf when compared to day-matched control oocytes. Subtracting the water-injected urea uptake or Pf values from those of UT-injected oocytes yielded UT-dependent values*. We demonstrate for the first time that UT-A2 and UT-A3 can transport water, and we confirm that UT-B is permeable to water. Moreover, the [14C] urea*/Pf* ratios fell in the sequence mUT-B>mUT-A2>mUT-A3, indicating that UTs can exhibit selectivity to urea and/or water. It is likely that specific kidney regions with high levels of UTs will exhibit increased urea and/or water permeabilities, directly influencing urine concentration. Furthermore, UT-mediated water transport activity must be considered when developing UT-inhibitors as novel diuretics. This article has an associated First Person interview with the first author of the paper.