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PMID: 9218515 Published · ppublish English Journal Article Research Support, U.S. Gov't, P.H.S.

Overexpression of the rat sarcoplasmic reticulum Ca2+ ATPase gene in the heart of transgenic mice accelerates calcium transients and cardiac relaxation.

The Journal of clinical investigation ·Vol. 100 ·No. 2 ·1997-07-15 ·Pages 380-9

He H, Giordano FJ, Hilal-Dandan R, Choi DJ, Rockman HA, McDonough PM, Bluhm WF, Meyer M, Sayen MR, Swanson E, Dillmann WH

Abstract

The Ca2+ ATPase of the sarcoplasmic reticulum (SERCA2) plays a dominant role in lowering cytoplasmic calcium levels during cardiac relaxation and reduction of its activity has been linked to delayed diastolic relaxation in hypothyroid and failing hearts. To determine the contractile alterations resulting from increased SERCA2 expression, we generated transgenic mice overexpressing a rat SERCA2 transgene. Characterization of a heterozygous transgenic mouse line (CJ5) showed that the amount of SERCA2 mRNA and protein increased 2. 6-fold and 1.2-fold, respectively, relative to control mice. Determination of the relative synthesis rate of SERCA2 protein showed an 82% increase. The mRNA levels of some of the other genes involved in calcium handling, such as the ryanodine receptor and calsequestrin, remained unchanged, but the mRNA levels of phospholamban and Na+/Ca2+ exchanger increased 1.4-fold and 1.8-fold, respectively. The increase in phospholamban or Na+/Ca2+ exchanger mRNAs did not, however, result in changes in protein levels. Functional analysis of calcium handling and contractile parameters in isolated cardiac myocytes indicated that the intracellular calcium decline (t1/2) and myocyte relengthening (t1/2) were accelerated by 23 and 22%, respectively. In addition, the rate of myocyte shortening was also significantly faster. In isolated papillary muscle from SERCA2 transgenic mice, the time to half maximum postrest potentiation was significantly shorter than in negative littermates. Furthermore, cardiac function measured in vivo, demonstrated significantly accelerated contraction and relaxation in SERCA2 transgenic mice that were further augmented in both groups with isoproterenol administration. Similar results were obtained for the contractile performance of myocytes isolated from a separate line (CJ2) of homozygous SERCA2 transgenic mice. Our findings suggest, for the first time, that increased SERCA2 expression is feasible in vivo and results in enhanced calcium transients, myocardial contractility, and relaxation that may have further therapeutic implications.

MeSH Terms
Animals Blotting, Northern Blotting, Southern Blotting, Western Calcium/metabolism Calcium-Transporting ATPases/genetics,metabolism Cells, Cultured Fluorescent Dyes/metabolism Gene Expression Regulation, Enzymologic Hemodynamics Indoles/metabolism Mice Mice, Transgenic Myocardial Contraction Myocardium/enzymology RNA, Messenger/metabolism Rats Sarcoplasmic Reticulum/enzymology Transgenes Ventricular Function
Chemicals
Fluorescent Dyes Indoles RNA, Messenger Calcium-Transporting ATPases indo-1 Calcium
Authors & Affiliations
11 authors, click to expand affiliations / ORCID
He H
Department of Medicine, University of California, San Diego, La Jolla, California 92093-0618, USA.
Giordano F J
Hilal-Dandan R
Choi D J
Rockman H A
McDonough P M
Bluhm W F
Meyer M
Sayen M R
Swanson E
Dillmann W H
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Article Info
Journal
The Journal of clinical investigation
Abbr.
J Clin Invest
ISSN
0021-9738
DOI
10.1172/JCI119544
Published
1997-07-15
Pages
380-9
Language
English
Region
United States
NLM ID
7802877
PMCID
PMC508201
Subset
IM
Grants
NHLBI NIH HHS · HL03041 · United States
NHLBI NIH HHS · HL0744-14 · United States
NHLBI NIH HHS · HL52946 · United States
External Links
PubMed source DOI: 10.1172/JCI119544 PMC Full Text View journal details
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