Localization and rapid regulation of Na+/myo-inositol cotransporter in rat kidney.
myo-inositol, a major compatible osmolyte in renal medulla, is accumulated in several kinds of cells under hypertonic conditions via Na+/myo-inositol cotransporter (SMIT). To investigate the physiological role of the SMIT, we sought to determine its localization by in situ hybridization and its acute regulation by NaCl and furosemide administration. Northern analysis demonstrated that SMIT is strongly expressed in the medulla and at low levels in the cortex of kidney. Intraperitoneal injection of NaCl rapidly induced SMIT mRNA in both the cortex and medulla, and furosemide completely abolished this induction. In situ hybridization revealed that SMIT it predominantly present in the medullary and cortical thick ascending limbs of Henle's loop (TALH) and macula densa cells. Less intense signals were seen in the inner medullary collecting ducts (IMCD). NaCl loading increased the signals throughout the TALH, and furosemide reduced the signals. SMIT in the IMCD is less sensitive to these kinds of acute regulation. Thus, the distribution pattern of SMIT does not correspond to the corticomedullary osmotic gradient, and SMIT in the TALH and macula densa cells is regulated very rapidly. These results suggest that SMIT expression in TALH may be regulated by intracellular and/or peritubular tonicity close to the basolateral membrane, which is supposed to be proportional to the magnitude of NaCl reabsorption.
Yamauchi
A
A
First Department of Medicine, Osaka University School of Medicine, Japan.
Miyai
A
A
Shimada
S
S
Minami
Y
Y
Tohyama
M
M
Imai
E
E
Kamada
T
T
Ueda
N
N
D000818:Animals; D001483:Base Sequence; D015152:Blotting, Northern; D002352:Carrier Proteins / Q000032:analysis / Q000096:biosynthesis / Q000378:metabolism*; D017931:DNA Primers; D005665:Furosemide / Q000494:pharmacology; D015870:Gene Expression / Q000187:drug effects; D006360:Heat-Shock Proteins / Q000032:analysis / Q000096:biosynthesis / Q000378:metabolism*; D017403:In Situ Hybridization; D007668:Kidney / Q000187:drug effects / Q000378:metabolism*; D007672:Kidney Cortex / Q000378:metabolism; D007679:Kidney Medulla / Q000378:metabolism; D007700:Kinetics; D008297:Male; D008565:Membrane Proteins; D008969:Molecular Sequence Data; D015345:Oligonucleotide Probes; D009928:Organ Specificity; D012333:RNA, Messenger / Q000032:analysis / Q000096:biosynthesis; D051381:Rats; D017208:Rats, Wistar; D027981:Symporters; D013997:Time Factors
D002352:Carrier Proteins; D017931:DNA Primers; D006360:Heat-Shock Proteins; D008565:Membrane Proteins; D015345:Oligonucleotide Probes; D012333:RNA, Messenger; D027981:Symporters; C073884:SLC5A3 protein, human; D005665:Furosemide