Phorbol 12-myristate 13-acetate alters SR Ca2+-ATPase gene expression in cultured neonatal rat heart cells

Q. I. Ming, José W M Bassani, Donald M Bers, Allen M. Samarel

Research output: Contribution to journalArticle

33 Citations (Scopus)

Abstract

-Primary cultures of neonatal rat ven-tricular myocytes were used to examine how the cardiac myocyte cytoplasmic Ca2+ ([Ca2+]i) transient and sarcoplasmic reticulum Ca2+-ATPase (SERCA2) gene expression change in response to treatment with the protein kinase C activator phorbol 12-myristate 13-acetate (PMA). Exposure of neonatal myocytes to PMA (200 nM, 48-72 h) produced myocyte growth and a 70% prolongation of the half-time for [Ca2+]i decline induced by potassium depolarization in the absence of extracellular Na+ (in which the sarcoplasmic reticulum Ca2+ pump is the main mechanism responsible for [Ca2+]i decline). The reduced rate of [Ca2+]i transient decline corresponded to a 53% reduction in SERCA2 protein levels and a 43% reduction in SERCA2 mRNA levels as compared with control myocytes. Exposure to PMA for as little as 30 min or for as long as 48 h produced a similar degree of SERCA2 mRNA downregulation over time. PMA-induced downregulation of SERCA2 mRNA levels was blocked by either 10 nM staurosporine or 4 uM chelerythrine, whereas treatment with either agent alone increased SERCA2 mRNA levels as compared with control cells. Actinomycin D mRNA stability assays revealed that PMA treatment appeared to markedly destabilize the relatively long-lived SERCA2 mRNA transcript. Taken together, these results indicate that downregulation of SERCA2 gene by PMA in cultured neonatal myocytes occurs at least in part by alterations in mRNA stability and results in functional alterations in [Ca2+]i decline that are similar to that observed in the hypertrophied and failing adult myocardium.

Original languageEnglish (US)
JournalAmerican Journal of Physiology
Volume271
Issue number3 PART 2
StatePublished - 1996
Externally publishedYes

Fingerprint

Calcium-Transporting ATPases
Acetates
Muscle Cells
Gene Expression
Messenger RNA
Down-Regulation
RNA Stability
Sarcoplasmic Reticulum
Staurosporine
Dactinomycin
Cardiac Myocytes
Protein Kinase C
phorbol-12-myristate
Myocardium
Potassium
Growth
Genes
Proteins

Keywords

  • Hypertrophy
  • Messenger ribonucleic acid stability
  • Myocardium
  • Protein kinase C
  • Sarcoplasmic reticulum

ASJC Scopus subject areas

  • Physiology (medical)

Cite this

Phorbol 12-myristate 13-acetate alters SR Ca2+-ATPase gene expression in cultured neonatal rat heart cells. / Ming, Q. I.; Bassani, José W M; Bers, Donald M; Samarel, Allen M.

In: American Journal of Physiology, Vol. 271, No. 3 PART 2, 1996.

Research output: Contribution to journalArticle

@article{5196ec9ffb1f47ddba58ab938f241d08,
title = "Phorbol 12-myristate 13-acetate alters SR Ca2+-ATPase gene expression in cultured neonatal rat heart cells",
abstract = "-Primary cultures of neonatal rat ven-tricular myocytes were used to examine how the cardiac myocyte cytoplasmic Ca2+ ([Ca2+]i) transient and sarcoplasmic reticulum Ca2+-ATPase (SERCA2) gene expression change in response to treatment with the protein kinase C activator phorbol 12-myristate 13-acetate (PMA). Exposure of neonatal myocytes to PMA (200 nM, 48-72 h) produced myocyte growth and a 70{\%} prolongation of the half-time for [Ca2+]i decline induced by potassium depolarization in the absence of extracellular Na+ (in which the sarcoplasmic reticulum Ca2+ pump is the main mechanism responsible for [Ca2+]i decline). The reduced rate of [Ca2+]i transient decline corresponded to a 53{\%} reduction in SERCA2 protein levels and a 43{\%} reduction in SERCA2 mRNA levels as compared with control myocytes. Exposure to PMA for as little as 30 min or for as long as 48 h produced a similar degree of SERCA2 mRNA downregulation over time. PMA-induced downregulation of SERCA2 mRNA levels was blocked by either 10 nM staurosporine or 4 uM chelerythrine, whereas treatment with either agent alone increased SERCA2 mRNA levels as compared with control cells. Actinomycin D mRNA stability assays revealed that PMA treatment appeared to markedly destabilize the relatively long-lived SERCA2 mRNA transcript. Taken together, these results indicate that downregulation of SERCA2 gene by PMA in cultured neonatal myocytes occurs at least in part by alterations in mRNA stability and results in functional alterations in [Ca2+]i decline that are similar to that observed in the hypertrophied and failing adult myocardium.",
keywords = "Hypertrophy, Messenger ribonucleic acid stability, Myocardium, Protein kinase C, Sarcoplasmic reticulum",
author = "Ming, {Q. I.} and Bassani, {Jos{\'e} W M} and Bers, {Donald M} and Samarel, {Allen M.}",
year = "1996",
language = "English (US)",
volume = "271",
journal = "American Journal of Physiology - Renal Fluid and Electrolyte Physiology",
issn = "1931-857X",
publisher = "American Physiological Society",
number = "3 PART 2",

}

TY - JOUR

T1 - Phorbol 12-myristate 13-acetate alters SR Ca2+-ATPase gene expression in cultured neonatal rat heart cells

AU - Ming, Q. I.

AU - Bassani, José W M

AU - Bers, Donald M

AU - Samarel, Allen M.

PY - 1996

Y1 - 1996

N2 - -Primary cultures of neonatal rat ven-tricular myocytes were used to examine how the cardiac myocyte cytoplasmic Ca2+ ([Ca2+]i) transient and sarcoplasmic reticulum Ca2+-ATPase (SERCA2) gene expression change in response to treatment with the protein kinase C activator phorbol 12-myristate 13-acetate (PMA). Exposure of neonatal myocytes to PMA (200 nM, 48-72 h) produced myocyte growth and a 70% prolongation of the half-time for [Ca2+]i decline induced by potassium depolarization in the absence of extracellular Na+ (in which the sarcoplasmic reticulum Ca2+ pump is the main mechanism responsible for [Ca2+]i decline). The reduced rate of [Ca2+]i transient decline corresponded to a 53% reduction in SERCA2 protein levels and a 43% reduction in SERCA2 mRNA levels as compared with control myocytes. Exposure to PMA for as little as 30 min or for as long as 48 h produced a similar degree of SERCA2 mRNA downregulation over time. PMA-induced downregulation of SERCA2 mRNA levels was blocked by either 10 nM staurosporine or 4 uM chelerythrine, whereas treatment with either agent alone increased SERCA2 mRNA levels as compared with control cells. Actinomycin D mRNA stability assays revealed that PMA treatment appeared to markedly destabilize the relatively long-lived SERCA2 mRNA transcript. Taken together, these results indicate that downregulation of SERCA2 gene by PMA in cultured neonatal myocytes occurs at least in part by alterations in mRNA stability and results in functional alterations in [Ca2+]i decline that are similar to that observed in the hypertrophied and failing adult myocardium.

AB - -Primary cultures of neonatal rat ven-tricular myocytes were used to examine how the cardiac myocyte cytoplasmic Ca2+ ([Ca2+]i) transient and sarcoplasmic reticulum Ca2+-ATPase (SERCA2) gene expression change in response to treatment with the protein kinase C activator phorbol 12-myristate 13-acetate (PMA). Exposure of neonatal myocytes to PMA (200 nM, 48-72 h) produced myocyte growth and a 70% prolongation of the half-time for [Ca2+]i decline induced by potassium depolarization in the absence of extracellular Na+ (in which the sarcoplasmic reticulum Ca2+ pump is the main mechanism responsible for [Ca2+]i decline). The reduced rate of [Ca2+]i transient decline corresponded to a 53% reduction in SERCA2 protein levels and a 43% reduction in SERCA2 mRNA levels as compared with control myocytes. Exposure to PMA for as little as 30 min or for as long as 48 h produced a similar degree of SERCA2 mRNA downregulation over time. PMA-induced downregulation of SERCA2 mRNA levels was blocked by either 10 nM staurosporine or 4 uM chelerythrine, whereas treatment with either agent alone increased SERCA2 mRNA levels as compared with control cells. Actinomycin D mRNA stability assays revealed that PMA treatment appeared to markedly destabilize the relatively long-lived SERCA2 mRNA transcript. Taken together, these results indicate that downregulation of SERCA2 gene by PMA in cultured neonatal myocytes occurs at least in part by alterations in mRNA stability and results in functional alterations in [Ca2+]i decline that are similar to that observed in the hypertrophied and failing adult myocardium.

KW - Hypertrophy

KW - Messenger ribonucleic acid stability

KW - Myocardium

KW - Protein kinase C

KW - Sarcoplasmic reticulum

UR - http://www.scopus.com/inward/record.url?scp=33745380416&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=33745380416&partnerID=8YFLogxK

M3 - Article

C2 - 8853338

AN - SCOPUS:33745380416

VL - 271

JO - American Journal of Physiology - Renal Fluid and Electrolyte Physiology

JF - American Journal of Physiology - Renal Fluid and Electrolyte Physiology

SN - 1931-857X

IS - 3 PART 2

ER -