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PMID: 9065779 Published · ppublish English Comparative Study Journal Article Research Support, Non-U.S. Gov't

Isoform diversity of the inositol trisphosphate receptor in cell types of mouse origin.

The Biochemical journal ·Vol. 322 ( Pt 2) ·1997-03-01 ·Pages 575-83

De Smedt H, Missiaen L, Parys JB, Henning RH, Sienaert I, Vanlingen S, Gijsens A, Himpens B, Casteels R

Abstract

Previous reports suggested the expression of four or five different Ins(1,4,5)P3 receptor [Ins(1,4,5)P3R] isoforms in mouse cells [Ross, Danoff, Schell, Snyder and Ullrich (1992) Proc. Natl. Acad. Sci. U.S.A. 89, 4265-4269; De Smedt, Missiaen, Parys, Bootman, Mertens, Van Den Bosch and Casteels (1994) J. Biol. Chem. 269, 21691-21698]. To explore this diversity further, we have isolated and sequenced partial clones of two Ins(1,4,5)P3R mRNAs from the mouse embryonic C3H10T1/2 cell line. These clones showed between 94.2 and 94.9% sequence identity with the corresponding rat Ins(1,4,5)P3R-II and Ins(1,4,5)P3R-III isoforms. Based on these newly obtained sequences we have determined the relative expression of the different Ins(1,4,5)P3R mRNAs in cultured cells and in animal tissues of mouse origin by a ratio reverse transcriptase polymerase chain reaction (RT-PCR). Ins(1,4,5)P3R-I was very prominent in brain and cerebellum and Ins(1,4,5)P3R-II in epithelia such as kidney as well as in both cardiac and skeletal muscle. Ins(1,4,5)P3R-III was highly expressed in all cultured cell types and in tissues with high cell turnover, e.g. testis. The prominent expression of Ins(1,4,5)P3R-I and Ins(1,4,5)P3R-III in A7r5 and C3H10T1/2 cells respectively was confirmed by immunoblot analysis and was compatible with a lower threshold for Ins(1,4,5)P3-induced Ca2+ release in the former cell type. Screening of a large number of mouse cell lines and tissues revealed the presence of Ins(1,4,5)P3R-I as well as of the Ins(1,4,5)P3R-II and Ins(1,4,5)P3R-III isoforms which were identified in the present study, but in contrast with previous reports there was no evidence for more isoform diversity.

MeSH Terms
Animals Base Sequence Calcium/metabolism Calcium Channels/classification,genetics Cell Line Cerebellum/chemistry Cloning, Molecular DNA Primers Embryo, Mammalian/cytology Genetic Variation Inositol 1,4,5-Trisphosphate Inositol 1,4,5-Trisphosphate Receptors Mice Microsomes/chemistry Molecular Sequence Data Polymerase Chain Reaction RNA-Directed DNA Polymerase Receptors, Cytoplasmic and Nuclear/classification,genetics Sequence Analysis, DNA Sequence Homology, Nucleic Acid Tissue Distribution
Chemicals
Calcium Channels DNA Primers Inositol 1,4,5-Trisphosphate Receptors Receptors, Cytoplasmic and Nuclear Inositol 1,4,5-Trisphosphate RNA-Directed DNA Polymerase Calcium
Authors & Affiliations
9 authors, click to expand affiliations / ORCID
De Smedt H
Laboratory of Physiology, Campus Gasthuisberg O/N, K.U. Leuven, Belgium.
Missiaen L
Parys J B
Henning R H
Sienaert I
Vanlingen S
Gijsens A
Himpens B
Casteels R
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Article Info
Journal
The Biochemical journal
Abbr.
Biochem J
ISSN
0264-6021
DOI
10.1042/bj3220575
Published
1997-03-01
Pages
575-83
Language
English
Region
England
NLM ID
2984726R
PMCID
PMC1218228
Subset
IM
Databases
GENBANK
Z71173, Z71174
External Links
PubMed source DOI: 10.1042/bj3220575 PMC Full Text View journal details
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