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

J Am Soc Nephrol. 2003 Sep;14(9):2229-36. doi: 10.1097/01.asn.0000085023.73801.4a.

Abstract

Chronic metabolic acidosis enhances the ability of the medullary thick ascending limb (MTAL) to absorb NH(4)(+) at least in part by stimulating the mRNA and protein expression of BSC1/NKCC2, the MTAL apical Na(+)-K(+)(NH(4)(+))-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. rBSC1 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 full-length mBSC1 cDNA but not in cells transfected with a mBSC1 cDNA lacking the 3'-UTR. These results demonstrate that acid pH enhances the stability of BSC1 mRNA probably by activating pathways that act on the AU-rich 3'-UTR of BSC1 mRNA, which contributes to the renal response to metabolic acidosis.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Acidosis / genetics
  • Acidosis / physiopathology*
  • Animals
  • Cells, Cultured
  • Hydrogen-Ion Concentration
  • In Vitro Techniques
  • Kidney Medulla / physiopathology*
  • Kidney Tubules / physiopathology
  • Mice
  • RNA Stability / genetics*
  • RNA, Messenger / genetics*
  • Rats
  • Sodium-Potassium-Chloride Symporters / genetics*
  • Solute Carrier Family 12, Member 1
  • Transcription, Genetic / genetics

Substances

  • RNA, Messenger
  • Slc12a1 protein, mouse
  • Slc12a1 protein, rat
  • Sodium-Potassium-Chloride Symporters
  • Solute Carrier Family 12, Member 1