Change in Nox4 expression is accompanied by changes in myogenic marker expression in differentiating C2C12 myoblasts

S Acharya, AM Peters, AS Norton, GK Murdoch, RA Hill

Research output: Contribution to journalArticle

8 Citations (Scopus)

Abstract

Myoblast differentiation is mediated by a cascade of changes in gene expression including transcription factors such as myogenin. Subsequent to myoblast differentiation, there is an increase in expression of the transmembrane protein NADPH oxidase (Nox). Nox is one of the primary factors for the generation of reactive oxygen species (ROS) in myogenic (C2C12) cells. Recently, ROS have been shown to be important regulators of several intracellular signaling pathways, and the full extent of their regulatory roles is yet to be discovered. In the present study, qRT PCR analysis demonstrated that Nox4 isoform is primarily expressed in differentiating C2C12 cells and contributes to the generation of ROS in C2C12 myoblast during differentiation. Over-expression and silencing of Nox4 expression during myoblast differentiation was accompanied by a reduction in intracellular ROS concentrations and an alteration in the expression patterns of Myf5, Pax7, MyoD1, and myogenin. This modulation was found to be associated with ERK1/2 phosphorylation. In both over-expression and reduced expression of Nox4, we found significant reductions in ERK1/2 phosphorylation. This indicates that cellular differentiation may be affected by Nox4-mediated endogenous ROS generation. These data suggest a new opportunity to study the temporal expression of Nox4 in the generation of ROS accompanying changes in myogenic differentiation.
Original languageEnglish
Pages (from-to)1181-1196
Number of pages16
JournalPflugers Archiv European Journal of Physiology
Volume465
Issue number8
DOIs
Publication statusPublished - 2013

Fingerprint

Myoblasts
Reactive Oxygen Species
Myogenin
Phosphorylation
NADPH Oxidase
Gene expression
Protein Isoforms
Transcription Factors
Modulation
Gene Expression
Polymerase Chain Reaction
Proteins

Cite this

@article{700192e7226c4744acb8b016c51fb684,
title = "Change in Nox4 expression is accompanied by changes in myogenic marker expression in differentiating C2C12 myoblasts",
abstract = "Myoblast differentiation is mediated by a cascade of changes in gene expression including transcription factors such as myogenin. Subsequent to myoblast differentiation, there is an increase in expression of the transmembrane protein NADPH oxidase (Nox). Nox is one of the primary factors for the generation of reactive oxygen species (ROS) in myogenic (C2C12) cells. Recently, ROS have been shown to be important regulators of several intracellular signaling pathways, and the full extent of their regulatory roles is yet to be discovered. In the present study, qRT PCR analysis demonstrated that Nox4 isoform is primarily expressed in differentiating C2C12 cells and contributes to the generation of ROS in C2C12 myoblast during differentiation. Over-expression and silencing of Nox4 expression during myoblast differentiation was accompanied by a reduction in intracellular ROS concentrations and an alteration in the expression patterns of Myf5, Pax7, MyoD1, and myogenin. This modulation was found to be associated with ERK1/2 phosphorylation. In both over-expression and reduced expression of Nox4, we found significant reductions in ERK1/2 phosphorylation. This indicates that cellular differentiation may be affected by Nox4-mediated endogenous ROS generation. These data suggest a new opportunity to study the temporal expression of Nox4 in the generation of ROS accompanying changes in myogenic differentiation.",
keywords = "NADPH oxidase Myogenin Differentiation Reactive oxygen species Mitogen-activated protein kinase MUSCLE PRECURSOR CELLS SKELETAL-MUSCLE REACTIVE OXYGEN NADPH OXIDASE OXIDATIVE STRESS ENDOTHELIAL-CELLS GENE-EXPRESSION SATELLITE CELLS ANGIOTENSIN-II UP-REGULATION Physiology PHYSIOLOGY Physiology",
author = "S Acharya and AM Peters and AS Norton and GK Murdoch and RA Hill",
note = "Cited References Count:63|195QW|SPRINGER|233 SPRING ST, NEW YORK, NY 10013 USA|Acharya, S.|Peters, A. M.|Norton, A. S.|Murdoch, G. K.|Hill, R. A.|ISI Document Delivery No.:195QW",
year = "2013",
doi = "10.1007/s00424-013-1241-0",
language = "English",
volume = "465",
pages = "1181--1196",
journal = "Pfluegers Archiv: European journal of physiology",
issn = "0031-6768",
publisher = "Springer-Verlag London Ltd.",
number = "8",

}

Change in Nox4 expression is accompanied by changes in myogenic marker expression in differentiating C2C12 myoblasts. / Acharya, S; Peters, AM; Norton, AS; Murdoch, GK; Hill, RA.

In: Pflugers Archiv European Journal of Physiology, Vol. 465, No. 8, 2013, p. 1181-1196.

Research output: Contribution to journalArticle

TY - JOUR

T1 - Change in Nox4 expression is accompanied by changes in myogenic marker expression in differentiating C2C12 myoblasts

AU - Acharya, S

AU - Peters, AM

AU - Norton, AS

AU - Murdoch, GK

AU - Hill, RA

N1 - Cited References Count:63|195QW|SPRINGER|233 SPRING ST, NEW YORK, NY 10013 USA|Acharya, S.|Peters, A. M.|Norton, A. S.|Murdoch, G. K.|Hill, R. A.|ISI Document Delivery No.:195QW

PY - 2013

Y1 - 2013

N2 - Myoblast differentiation is mediated by a cascade of changes in gene expression including transcription factors such as myogenin. Subsequent to myoblast differentiation, there is an increase in expression of the transmembrane protein NADPH oxidase (Nox). Nox is one of the primary factors for the generation of reactive oxygen species (ROS) in myogenic (C2C12) cells. Recently, ROS have been shown to be important regulators of several intracellular signaling pathways, and the full extent of their regulatory roles is yet to be discovered. In the present study, qRT PCR analysis demonstrated that Nox4 isoform is primarily expressed in differentiating C2C12 cells and contributes to the generation of ROS in C2C12 myoblast during differentiation. Over-expression and silencing of Nox4 expression during myoblast differentiation was accompanied by a reduction in intracellular ROS concentrations and an alteration in the expression patterns of Myf5, Pax7, MyoD1, and myogenin. This modulation was found to be associated with ERK1/2 phosphorylation. In both over-expression and reduced expression of Nox4, we found significant reductions in ERK1/2 phosphorylation. This indicates that cellular differentiation may be affected by Nox4-mediated endogenous ROS generation. These data suggest a new opportunity to study the temporal expression of Nox4 in the generation of ROS accompanying changes in myogenic differentiation.

AB - Myoblast differentiation is mediated by a cascade of changes in gene expression including transcription factors such as myogenin. Subsequent to myoblast differentiation, there is an increase in expression of the transmembrane protein NADPH oxidase (Nox). Nox is one of the primary factors for the generation of reactive oxygen species (ROS) in myogenic (C2C12) cells. Recently, ROS have been shown to be important regulators of several intracellular signaling pathways, and the full extent of their regulatory roles is yet to be discovered. In the present study, qRT PCR analysis demonstrated that Nox4 isoform is primarily expressed in differentiating C2C12 cells and contributes to the generation of ROS in C2C12 myoblast during differentiation. Over-expression and silencing of Nox4 expression during myoblast differentiation was accompanied by a reduction in intracellular ROS concentrations and an alteration in the expression patterns of Myf5, Pax7, MyoD1, and myogenin. This modulation was found to be associated with ERK1/2 phosphorylation. In both over-expression and reduced expression of Nox4, we found significant reductions in ERK1/2 phosphorylation. This indicates that cellular differentiation may be affected by Nox4-mediated endogenous ROS generation. These data suggest a new opportunity to study the temporal expression of Nox4 in the generation of ROS accompanying changes in myogenic differentiation.

KW - NADPH oxidase Myogenin Differentiation Reactive oxygen species Mitogen-activated protein kinase MUSCLE PRECURSOR CELLS SKELETAL-MUSCLE REACTIVE OXYGEN NADPH OXIDASE OXIDATIVE STRESS ENDOTHELIAL-CELLS GENE-EXPRESSION SATELLITE CELLS ANGIOTENSIN-II UP-REGUL

U2 - 10.1007/s00424-013-1241-0

DO - 10.1007/s00424-013-1241-0

M3 - Article

C2 - 23503725

VL - 465

SP - 1181

EP - 1196

JO - Pfluegers Archiv: European journal of physiology

JF - Pfluegers Archiv: European journal of physiology

SN - 0031-6768

IS - 8

ER -