PMID- 21057045
OWN - NLM
STAT- MEDLINE
DCOM- 20110128
LR  - 20211020
IS  - 1522-1539 (Electronic)
IS  - 0363-6135 (Print)
IS  - 0363-6135 (Linking)
VI  - 300
IP  - 1
DP  - 2011 Jan
TI  - Silencing calcineurin A subunit reduces SERCA2 expression in cardiac myocytes.
PG  - H173-80
LID - 10.1152/ajpheart.00841.2010 [doi]
AB  - Resting intracellular Ca(2+) can be raised, in neonatal rat cardiac myocytes, by 
      exposure to very low concentration of thapsigargin (TG). Such a Ca(2+) rise 
      yields calcineurin (CN) activation demonstrated by increased expression of 
      transfected luciferase cDNA under control of nuclear factor of activated T-cells 
      (NFAT) promoter and increased translocation of NFAT to nuclei. We found that 
      exposure of cardiac myocytes to TG is followed by increase of sarcroplasmic 
      reticulum Ca(2+) transport ATPase (SERCA2) expression, which is further increased 
      when CN inactivation by CAMKII (calmodulin-dependent kinase) is prevented with 
      KN93 (CAMKII inhibitor). On the other hand, SERCA2 expression is reduced by CN 
      inhibition with cyclosporine. We have now induced calcineurin A (CNA) alpha- or 
      beta-subunit gene silencing with small interfering RNA (siRNA) and observed strong 
      interference with expression of SERCA2, both in control myocytes and following 
      exposure to TG. Such interference is also obtained following NFAT displacement 
      from CN with 9,10-dihydro-9,10[1',2']-benzenoanthracene-1,4-dione (INCA-6). We 
      have also observed analogous effects on expression of phospholamban (PLB) and 
      Na(+)/Ca(2+) exchanger (NCX). Pertinent to these findings, we have identified, by 
      in-silico analysis, NFAT binding sites in SERCA2, PLB, and NCX1 promoters. Our 
      experiments indicate that activation of the calcineurin-NFAT pathway by rise of 
      resting cytosolic Ca(2+) elevates transcription/expression of SERCA2, PLB, and 
      NCX1, providing a homeostatic mechanism for long-term control of cytosolic 
      Ca(2+).
FAU - Prasad, Anand Mohan
AU  - Prasad AM
AD  - California Pacific Medical Center Research Institute, San Francisco, California 
      94107, USA.
FAU - Inesi, Giuseppe
AU  - Inesi G
LA  - eng
GR  - R01-69830/PHS HHS/United States
PT  - Journal Article
PT  - Research Support, N.I.H., Extramural
DEP - 20101105
PL  - United States
TA  - Am J Physiol Heart Circ Physiol
JT  - American journal of physiology. Heart and circulatory physiology
JID - 100901228
RN  - 0 (RNA, Small Interfering)
RN  - 67526-95-8 (Thapsigargin)
RN  - EC 3.1.3.16 (Calcineurin)
RN  - EC 3.6.3.8 (Sarcoplasmic Reticulum Calcium-Transporting ATPases)
RN  - SY7Q814VUP (Calcium)
SB  - IM
MH  - Analysis of Variance
MH  - Animals
MH  - Blotting, Western
MH  - Calcineurin/*genetics/metabolism
MH  - Calcium/*metabolism
MH  - Cells, Cultured
MH  - Fluorescent Antibody Technique
MH  - Myocytes, Cardiac/drug effects/*metabolism
MH  - RNA, Small Interfering
MH  - Rats
MH  - Reverse Transcriptase Polymerase Chain Reaction
MH  - Sarcoplasmic Reticulum/drug effects/metabolism
MH  - Sarcoplasmic Reticulum Calcium-Transporting ATPases/genetics/*metabolism
MH  - Thapsigargin/pharmacology
MH  - Up-Regulation
PMC - PMC3023267
EDAT- 2010/11/09 06:00
MHDA- 2011/02/01 06:00
PMCR- 2012/01/01
CRDT- 2010/11/09 06:00
PHST- 2010/11/09 06:00 [entrez]
PHST- 2010/11/09 06:00 [pubmed]
PHST- 2011/02/01 06:00 [medline]
PHST- 2012/01/01 00:00 [pmc-release]
AID - ajpheart.00841.2010 [pii]
AID - H-00841-2010 [pii]
AID - 10.1152/ajpheart.00841.2010 [doi]
PST - ppublish
SO  - Am J Physiol Heart Circ Physiol. 2011 Jan;300(1):H173-80. doi: 
      10.1152/ajpheart.00841.2010. Epub 2010 Nov 5.