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J Biol Chem, Vol. 273, Issue 50, 33681-33691, December 11, 1998

Stimulation by in Vivo and in Vitro Metabolic Acidosis of Expression of rBSC-1, the Na⁺-K⁺(NH₄⁺)-2Cl Cotransporter of the Rat Medullary Thick Ascending Limb

Amel Attmane-Elakeb, David B. Mount^§, Valérie Sibella, Catherine Vernimmen, Steven C. Hebert^§, and Maurice Bichara

From  INSERM Unité 356, Physiologie et Endocrinologie Cellulaire et Moléculaire Rénale, Université Pierre et Marie Curie, 75006 Paris, France and the ^§ Division of Nephrology, Vanderbilt University, Nashville, Tennessee 37232-2372

To assess whether metabolic acidosis per se regulates rBSC-1, the rat medullary thick ascending limb (MTAL) apical Na⁺-K⁺(NH₄⁺)-2Cl cotransporter, rat MTALs were incubated for 16 h in an acid 1:1 mixture of Ham's nutrient mixture F-12 and Dulbecco's modified Eagle's medium. Cotransport activity was estimated in intact cells and membrane vesicles by intracellular pH and ²²Na⁺ uptake measurements, respectively; rBSC-1 protein was quantified by immunoblotting analysis and mRNA by quantitative reverse transcription-polymerase chain reaction. As compared with incubation at pH ~7.35, acid incubation (pH ~7.10) up-regulated by 35-100% rBSC-1 transport activity in cells and membrane vesicles, and rBSC-1 protein and mRNA abundance. In contrast, acid incubation did not alter alkaline phosphatase and Na⁺/K⁺-ATPase enzyme activities or -actin protein abundance. After 3 h of in vivo chronic metabolic acidosis (CMA) rBSC-1 mRNA abundance increased in freshly harvested MTALs, which was accompanied after 1-6 days of CMA with enhanced rBSC-1 protein abundance. These results demonstrate that both in vivo and in vitro CMA stimulate rBSC-1 expression, which would contribute to the adaptive increase in MTAL absorption and urinary excretion of NH₄⁺ in response to CMA.

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