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.

作者: Amel Attmane-Elakeb , David B. Mount , Valérie Sibella , Catherine Vernimmen , Steven C. Hebert

DOI: 10.1074/JBC.273.50.33681

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

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