Acid pH increases the stability of BSC1/NKCC2 mRNA in the medullary thick ascending limb

Chronic metabolic acidosis enhances the ability of the medullary thick ascending limb (MTAL) to absorb NH4+ at least in part by stimulating the mRNA and protein expression of BSC1/NKCC2., the MTAL apical Na+-K+(NH4+)-2Cl(-)co-transporter. For assessing the mechanism by which an acid pH enhances the BSC1 mRNA abundance, MTAL were harvested from adrenalectomized rats and incubated in control (pH 7.35) and acid (pH 7.10) 1:1 mixtures of Ham's nutrient mixture F-12 and DME. rBSCl mRNA abundance and gene transcription rate were quantified by quantitative reverse transcription-PCR and run-off assay, respectively. Acid incubation enhanced mRNA abundance within 4 h in whole cell (P < 0.02) but not in nucleus. BSC1 gene transcription rate was not affected by acid incubation. In contrast, under conditions in which gene transcription was blocked, rBSC1 mRNA decreased within 6 h by 38 +/- 11% in control but only by 15 +/- 15% in acid medium (P < 0.02), which represented an increase in the BSC1 mRNA half-life from approximately 7 to approximately 17 h. Furthermore, in a mouse TAL cell line, acid incubation for 16 h significantly increased (P < 0.02) the amount of BSC1 mRNA in cells transfected with the fulllength mBSC1 cDNA but not in cells transfected with a mBSC1 cDNA lacking the Y-UTR. These results demonstrate that acid pH enhances the stability of BSC1 mRNA probably by activating pathways that act on the AU-rich Y-UTR of BSC1 mRNA, which contributes to the renal response to metabolic acidosis.