Hypertonicity stimulates transcription of gene for Na(+)-myo-inositol cotransporter in MDCK cells.
Myo-inositol is a major compatible osmolyte accumulated in the hypertonic renal medulla and in Madin-Darby canine kidney (MDCK) cells cultured in hypertonic media. Myo-inositol is taken up by MDCK cells on a Na(+)-coupled transporter whose activity increases sixfold 24 h after cells are switched to hypertonic medium. To investigate the mechanism of regulation of the cotransporter by hypertonicity, we used the cDNA encoding the canine Na(+)-myo-inositol cotransporter that we recently cloned to measure the abundance of the mRNA for the cotransporter and its rate of transcription after changes in osmolality. When MDCK cells were switched from isotonic to hypertonic medium, cotransporter mRNA abundance rose 10-fold in 16 h. Transcription of the cotransporter gene also rose and 16 h after the switch reached a peak approximately 15-fold that in isotonic cells. When cells were switched back to isotonic medium, mRNA abundance and transcription of the gene returned to isotonic levels in 8 h and transport rate reached isotonic levels in 48 h. Thus transcription appears to be the primary step in regulation of myo-inositol transport by hypertonicity.
Yamauchi
A
A
Department of Medicine, Johns Hopkins University School of Medicine, Baltimore, Maryland 21205.
Uchida
S
S
Preston
A S
AS
Kwon
H M
HM
Handler
J S
JS
D000818:Animals; D015152:Blotting, Northern; D002352:Carrier Proteins / Q000235:genetics*; D002460:Cell Line; D003470:Culture Media; D004285:Dogs; D006360:Heat-Shock Proteins / Q000235:genetics*; D006982:Hypertonic Solutions / Q000494:pharmacology*; D008565:Membrane Proteins; D009693:Nucleic Acid Hybridization; D012333:RNA, Messenger / Q000378:metabolism; D012260:Ribonucleases; D027981:Symporters; D014158:Transcription, Genetic / Q000187:drug effects*
D002352:Carrier Proteins; D003470:Culture Media; D006360:Heat-Shock Proteins; D006982:Hypertonic Solutions; D008565:Membrane Proteins; D012333:RNA, Messenger; D027981:Symporters; C073884:SLC5A3 protein, human; D012260:Ribonucleases