Differential upregulation of rat Na-K-Cl cotransporter, rBSC1, mRNA in the thick ascending limb of Henle in different pathological conditions

Differential upregulation of rat Na-K-Cl cotransporter, rBSC1, mRNA in the thick ascending limb of Henle in different pathological conditions. Na-Cl cotransport across the apical membrane of epithelial cells in the thick ascending limb of the loop of Henle (TAL) plays a major role in salt accumulati...

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Published in:Kidney international 1998-09, Vol.54 (3), p.877-888
Main Authors: Marumo, Rie, Kaizuma, Sakiko, Nogae, Shoji, Kanazawa, Masayuki, Kimura, Tokihisa, Saito, Takao, Ito, Sadayoshi, Matsubara, Mitsunobu
Format: Article
Language:English
Subjects:
Animals
ANP
Arginine Vasopressin - pharmacology
AVP
Biological and medical sciences
cardiac failure
Carrier Proteins - genetics
CHF
cotransport
dehydration
Dehydration - metabolism
Gene Expression Regulation
Heart Failure - metabolism
In Situ Hybridization
Investigative techniques of renal and urinary tract function
Investigative techniques, diagnostic techniques (general aspects)
Loop of Henle - metabolism
Male
Medical sciences
osmolality
Rats
Rats, Sprague-Dawley
RNA, Messenger - analysis
sodium transporter
Sodium-Potassium-Chloride Symporters
transcription
Up-Regulation
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Summary:Differential upregulation of rat Na-K-Cl cotransporter, rBSC1, mRNA in the thick ascending limb of Henle in different pathological conditions. Na-Cl cotransport across the apical membrane of epithelial cells in the thick ascending limb of the loop of Henle (TAL) plays a major role in salt accumulation for hypertonic medullary interstitium. The electroneutral, rat bumetanide-sensitive sodium transporter, rBSC1, is involved in this process. We studied the level of rBSC1 mRNA in dehydration and cardiac failure, since sodium transport in TAL may be enhanced in both conditions in spite of the difference in extracellular fluid accumulation. Male Sprague-Dawley rats were deprived of water for 24hours and myocardial infarction of about 40% of left ventricular circumference was induced in another group of rats that later developed congestive heart failure (CHF). Digoxigenin-labeled cRNA probe for rBSC1 was constructed using polymerase chain reaction (PCR), and Northern blot analysis was performed using RNAs from renal outer medulla. By inducing a point mutation at the middle of PCR product, we compared the amount of rBSC1 transcripts in the renal cortex using competitive PCR, since TAL represents a small fraction of the total cortical tissue. Northern analysis showed a significant increase in rBSC1 mRNA in the renal outer medulla of both dehydrated and CHF rats. In the renal cortex, however, the increase was noted only in CHF by competitive PCR. In situ hybridization using the riboprobe for northern analysis demonstrated that the transcript signal in dehydrated rats was intensified segmentally in TAL located in the inner stripe of outer medulla. Western analysis and immunohistochemistry using a specific antibody against rBSC1 confirmed the distinct segmental enhancement of apical protein expression in dehydration and diffuse enhanced expression in CHF. rBSC1 is differentially upregulated in different pathological conditions.
ISSN:0085-2538
1523-1755
DOI:10.1046/j.1523-1755.1998.00051.x