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PMID: 15546997 Published · ppublish English Journal Article Research Support, U.S. Gov't, Non-P.H.S. Research Support, U.S. Gov't, P.H.S.

Sarco(endo)plasmic reticulum Ca2+ ATPase (SERCA) gene silencing and remodeling of the Ca2+ signaling mechanism in cardiac myocytes.

Proceedings of the National Academy of Sciences of the United States of America ·Vol. 101 ·No. 47 ·2004-11-23 ·Pages 16683-8

Seth M, Sumbilla C, Mullen SP, Lewis D, Klein MG, Hussain A, Soboloff J, Gill DL, Inesi G

Abstract

Transient elevations of cytosolic Ca2+ are a common mechanism of cellular signaling. In striated muscle, the sarco(endo)plasmic reticulum Ca2+ ATPase (SERCA) plays an important role in terminating Ca2+ transients by returning cytosolic Ca2+ to intracellular stores. Stored Ca2+ can then be released again for subsequent signaling. We down-regulated SERCA2 gene expression in cultured cardiac myocytes by means of endogenous transcription of small interfering RNA encoded by an exogenous cDNA template. The cDNA template was delivered by adenovirus vector. Reduction of SERCA expression in all myocytes in culture was documented by immunochemistry, real-time RT-PCR, and determination of ATP-dependent Ca2+ transport. The reduction of SERCA2 expression was associated with the up-regulation of transient receptor potential (TRP) channel proteins (TRPC4 and TRPC5) and Na+/Ca2+ exchanger, indicating that intracellular store deficiency was compensated for by Ca2+ fluxes through the plasma membrane. In fact, SERCA silencing was followed by increased transcription of Na+/Ca2+ exchanger, TRPC4, TRPC5, and related transcriptional factors, such as stimulating protein 1, myocyte enhancer factor 2, and nuclear factor of activated cells 4, through activation of calcineurin. This finding demonstrates that the observed compensation occurs through transcriptional crosstalk and the remodeling of Ca2+ signaling pathways. The wide significance of this regulatory mechanism is related to its general involvement in Ca2+ signaling dynamics and in cardiac development and hypertrophy.

MeSH Terms
Animals Base Sequence Calcium Signaling Calcium-Transporting ATPases/genetics,metabolism Cells, Cultured Chick Embryo Cricetinae Down-Regulation Gene Silencing Humans Ion Channels/metabolism Myocytes, Cardiac/metabolism RNA, Small Interfering/genetics Rats Sarcoplasmic Reticulum Calcium-Transporting ATPases
Chemicals
Atp2a2 protein, rat Ion Channels RNA, Small Interfering Sarcoplasmic Reticulum Calcium-Transporting ATPases ATP2A2 protein, human Calcium-Transporting ATPases
Authors & Affiliations
9 authors, click to expand affiliations / ORCID
Seth M
Department of Biochemistry and Molecular Biology, University of Maryland School of Medicine, Baltimore, MD 21201, USA.
Sumbilla C
Mullen S P
Lewis D
Klein M G
Hussain A
Soboloff J
Gill D L
Inesi G
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Article Info
Journal
Proceedings of the National Academy of Sciences of the United States of America
Abbr.
Proc Natl Acad Sci U S A
ISSN
0027-8424
DOI
10.1073/pnas.0407537101
Published
2004-11-23
Epub
2004-00-16
Pages
16683-8
Language
English
Region
United States
NLM ID
7505876
PMCID
PMC528906
Subset
IM
Grants
NHLBI NIH HHS · HL69830 · United States
NHLBI NIH HHS · R01 HL069830 · United States
NHLBI NIH HHS · HL55426 · United States
NIAID NIH HHS · AI058173 · United States
NIAID NIH HHS · R01 AI058173 · United States
NHLBI NIH HHS · R01 HL055426 · United States
External Links
PubMed source DOI: 10.1073/pnas.0407537101 PMC Full Text View journal details
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