Na+/H+-exchanger 3 activity and its gene in the spontaneously hypertensive rat kidney

Matsuhiko Hayashi, Tadashi Yoshida, Toshiaki Monkawa, Yasuyoshi Yamaji, Souichirou Sato, Takao Saruta

Research output: Contribution to journalArticle

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Abstract

Objective. To examine the mechanisms affecting proximal tubule sodium reabsorption, we studied the gene expression and functional changes of the Na+/H+-exchanger 3 (NHE3) and its gene expression in spontaneously hypertensive rats (SHR). Methods. The S1 segment of the superficial proximal convoluted tubules (PCT) was microdissected from collagenase-treated kidneys of SHR and Wistar-Kyoto (WKY) rats. The tubules were then superfused in a HEPES-buffered solution. The intracellular pH was measured by monitoring changes in fluorescent dye emission. By measuring the amiloride-sensitive intracellular pH recovery rate after intracellular acid loading with NH4Cl, we estimated the NHE3 activity. Northern blotting was performed to quantitate the NHE3 messenger RNA (mRNA) expression, and the NHE3 complementary DNA sequence was determined both in SHR and in WKY rat strains. Results. NHE3 activity was significantly higher in SHR than it was in WKY rats both at age 4 weeks and at age 7 weeks. Northern blot analysis showed that the mRNA expression did not differ significantly in the two strains at either age. Sequence analysis of complementary DNA of SHR NHE3 revealed five allelic variations compared with the known wild-type sequence in Sprague-Dawley rats. The same variations were also identified in the WKY rats. Conclusion. NHE3 activity is increased in SHR compared with WKY rats of the same age. Changes in the NHE3 mRNA expression level and protein structure are not responsible for this difference between SHR and WKY rat strains.

Original languageEnglish
Pages (from-to)43-48
Number of pages6
JournalJournal of Hypertension
Volume15
Issue number1
Publication statusPublished - 1997

Fingerprint

Sodium-Hydrogen Antiporter
Inbred SHR Rats
Inbred WKY Rats
Kidney
Genes
Northern Blotting
Messenger RNA
Complementary DNA
HEPES
Gene Expression
Amiloride
Collagenases
Fluorescent Dyes
Sequence Analysis
Sprague Dawley Rats
Sodium
Acids

Keywords

  • Complementary DNA
  • Messenger RNA
  • Na/H-exchanger 3
  • Spontaneously hypertensive rats
  • Wistar-Kyoto rats

ASJC Scopus subject areas

  • Internal Medicine
  • Endocrinology

Cite this

Na+/H+-exchanger 3 activity and its gene in the spontaneously hypertensive rat kidney. / Hayashi, Matsuhiko; Yoshida, Tadashi; Monkawa, Toshiaki; Yamaji, Yasuyoshi; Sato, Souichirou; Saruta, Takao.

In: Journal of Hypertension, Vol. 15, No. 1, 1997, p. 43-48.

Research output: Contribution to journalArticle

Hayashi, Matsuhiko ; Yoshida, Tadashi ; Monkawa, Toshiaki ; Yamaji, Yasuyoshi ; Sato, Souichirou ; Saruta, Takao. / Na+/H+-exchanger 3 activity and its gene in the spontaneously hypertensive rat kidney. In: Journal of Hypertension. 1997 ; Vol. 15, No. 1. pp. 43-48.
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abstract = "Objective. To examine the mechanisms affecting proximal tubule sodium reabsorption, we studied the gene expression and functional changes of the Na+/H+-exchanger 3 (NHE3) and its gene expression in spontaneously hypertensive rats (SHR). Methods. The S1 segment of the superficial proximal convoluted tubules (PCT) was microdissected from collagenase-treated kidneys of SHR and Wistar-Kyoto (WKY) rats. The tubules were then superfused in a HEPES-buffered solution. The intracellular pH was measured by monitoring changes in fluorescent dye emission. By measuring the amiloride-sensitive intracellular pH recovery rate after intracellular acid loading with NH4Cl, we estimated the NHE3 activity. Northern blotting was performed to quantitate the NHE3 messenger RNA (mRNA) expression, and the NHE3 complementary DNA sequence was determined both in SHR and in WKY rat strains. Results. NHE3 activity was significantly higher in SHR than it was in WKY rats both at age 4 weeks and at age 7 weeks. Northern blot analysis showed that the mRNA expression did not differ significantly in the two strains at either age. Sequence analysis of complementary DNA of SHR NHE3 revealed five allelic variations compared with the known wild-type sequence in Sprague-Dawley rats. The same variations were also identified in the WKY rats. Conclusion. NHE3 activity is increased in SHR compared with WKY rats of the same age. Changes in the NHE3 mRNA expression level and protein structure are not responsible for this difference between SHR and WKY rat strains.",
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AU - Sato, Souichirou

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N2 - Objective. To examine the mechanisms affecting proximal tubule sodium reabsorption, we studied the gene expression and functional changes of the Na+/H+-exchanger 3 (NHE3) and its gene expression in spontaneously hypertensive rats (SHR). Methods. The S1 segment of the superficial proximal convoluted tubules (PCT) was microdissected from collagenase-treated kidneys of SHR and Wistar-Kyoto (WKY) rats. The tubules were then superfused in a HEPES-buffered solution. The intracellular pH was measured by monitoring changes in fluorescent dye emission. By measuring the amiloride-sensitive intracellular pH recovery rate after intracellular acid loading with NH4Cl, we estimated the NHE3 activity. Northern blotting was performed to quantitate the NHE3 messenger RNA (mRNA) expression, and the NHE3 complementary DNA sequence was determined both in SHR and in WKY rat strains. Results. NHE3 activity was significantly higher in SHR than it was in WKY rats both at age 4 weeks and at age 7 weeks. Northern blot analysis showed that the mRNA expression did not differ significantly in the two strains at either age. Sequence analysis of complementary DNA of SHR NHE3 revealed five allelic variations compared with the known wild-type sequence in Sprague-Dawley rats. The same variations were also identified in the WKY rats. Conclusion. NHE3 activity is increased in SHR compared with WKY rats of the same age. Changes in the NHE3 mRNA expression level and protein structure are not responsible for this difference between SHR and WKY rat strains.

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