Intracellular calcium concentration in the inositol trisphosphate receptor type 1 knockout mouse

Matsuhiko Hayashi, Toshiaki Monkawa, Tadashi Yoshida, Hiroyuki Sasamura, Mineo Matsumoto, Takafumi Inoue, Katsuhiko Mikoshiba, Takao Saruta

Research output: Contribution to journalArticle

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Abstract

Recently, mice with a disrupted inositol trisphosphate (IP3) receptor type 1 allele were produced by gene targeting. To examine the role of IP3 receptor type 1 in the regulation of intracellular calcium concentration ([Ca2+](i)) of glomerular cells, [Ca2+](i) was measured with fura 2- acetoxymethyl-ester in the superfused glomeruli from homozygous and wild-type mice. [Ca2+](i) was determined in calcium-free medium before and after the addition of 10-7 M endothelin-1 (ET- 1) and 10-6 M angiotensin II (AngII). The expression of mRNA of IP3 receptor isoforms and hormone receptors in the glomeruli from these animals also was measured by quantitative reverse transcription-PCR with specific primers for IP3 receptor isoforms (types 1, 2, and 3), AngII receptor type 1, and ET receptors (types A and B). In homozygous mutants, the shorter mRNA of IP3 receptor type 1, which lacks the first exon, is transcribed. Basal [Ca2+](i) and the responses to ET-1 and AngII in homozygous mutants (ET-1, 55 ± 7 nM to 73 ± 7 nM; AngII, 66 ± 6 to 91 ± 8 nM) were significantly lower than those in the wild-type mice (ET-1, 93 ± 13 nM to 162 ± 13 nM; AngII, 87 ± 7 to 147 ± 9 nM; P < 0.05 for both hormones) without significant changes in mRNA expression of hormone receptors. The results with quantitative reverse transcription-PCR also revealed that mRNA expression of the IP3 receptor gene family was not significantly different between the two groups. The present study clearly shows that IP3 receptor type 1 plays a major role in the regulation of [Ca2+](i) in the glomeruli and that lack of an isoform of IP3 receptor in the glomeruli does not induce expression of the other isoforms of the IP3 receptor.

Original languageEnglish
Pages (from-to)2094-2101
Number of pages8
JournalJournal of the American Society of Nephrology
Volume10
Issue number10
Publication statusPublished - 1999 Oct
Externally publishedYes

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Inositol 1,4,5-Trisphosphate Receptors
Inositol
Knockout Mice
Calcium
Angiotensin II
Protein Isoforms
Messenger RNA
Hormones
Reverse Transcription
Polymerase Chain Reaction
Angiotensin Type 1 Receptor
Gene Targeting
Fura-2
Endothelin-1
Exons
Esters
Alleles

ASJC Scopus subject areas

  • Nephrology

Cite this

Hayashi, M., Monkawa, T., Yoshida, T., Sasamura, H., Matsumoto, M., Inoue, T., ... Saruta, T. (1999). Intracellular calcium concentration in the inositol trisphosphate receptor type 1 knockout mouse. Journal of the American Society of Nephrology, 10(10), 2094-2101.

Intracellular calcium concentration in the inositol trisphosphate receptor type 1 knockout mouse. / Hayashi, Matsuhiko; Monkawa, Toshiaki; Yoshida, Tadashi; Sasamura, Hiroyuki; Matsumoto, Mineo; Inoue, Takafumi; Mikoshiba, Katsuhiko; Saruta, Takao.

In: Journal of the American Society of Nephrology, Vol. 10, No. 10, 10.1999, p. 2094-2101.

Research output: Contribution to journalArticle

Hayashi, M, Monkawa, T, Yoshida, T, Sasamura, H, Matsumoto, M, Inoue, T, Mikoshiba, K & Saruta, T 1999, 'Intracellular calcium concentration in the inositol trisphosphate receptor type 1 knockout mouse', Journal of the American Society of Nephrology, vol. 10, no. 10, pp. 2094-2101.
Hayashi, Matsuhiko ; Monkawa, Toshiaki ; Yoshida, Tadashi ; Sasamura, Hiroyuki ; Matsumoto, Mineo ; Inoue, Takafumi ; Mikoshiba, Katsuhiko ; Saruta, Takao. / Intracellular calcium concentration in the inositol trisphosphate receptor type 1 knockout mouse. In: Journal of the American Society of Nephrology. 1999 ; Vol. 10, No. 10. pp. 2094-2101.
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