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PMID: 15774533 Published · ppublish English Journal Article Research Support, N.I.H., Extramural Research Support, U.S. Gov't, P.H.S.

Mitochondria-associated apoptotic signalling in denervated rat skeletal muscle.

The Journal of physiology ·Vol. 565 ·No. Pt 1 ·2005-05-15 ·Pages 309-23

Siu PM, Alway SE

Abstract

Apoptosis has been implicated in the regulation of denervation-induced muscle atrophy. However, the activation of apoptotic signal transduction during muscle denervation has not been fully elucidated. The present study examined the apoptotic responses to denervation in rat gastrocnemius muscle. Following 14 days of denervation, the extent of apoptotic DNA fragmentation as determined by a cytosolic nucleosome ELISA was increased by 100% in the gastrocnemius muscle. RT-PCR and immunoblot analyses indicated that Bax was dramatically upregulated while Bcl-2 was modestly increased; however, the Bax/Bcl-2 ratio was significantly increased in denervated muscles relative to control muscles. Analyses of ELISA and immunoblots from mitochondria-free cytosol extracts showed a significant increase in mitochondria-associated apoptotic factors, including cytochrome c, Smac/DIABLO and apoptosis-inducing factor (AIF). In addition to the upregulation of caspase-3 and -9 mRNA, pro-/cleaved caspase protein and proteolytic activity levels, the X-linked inhibitor of apoptosis (XIAP) protein level was downregulated. The cleaved product of poly(ADP-ribose) polymerase (PARP) was detected in muscle samples following denervation. Although we did not find a difference in the inhibitor of DNA binding/differentiation-2 (Id2) and c-Myc protein contents between the denervated and control muscles, the protein content of tumour suppressor p53 was significantly increased in both the nuclear and the cytosolic fractions with denervation. Moreover, denervation increased the protein content of HSP70, whereas the MnSOD (a mitochondrial isoform of superoxide dismutase) protein content was diminished, which indicated that denervation might have induced cellular and/or oxidative stress. Our data show that mitochondria-associated apoptotic signalling is upregulated during muscle denervation. We interpret these findings to indicate that apoptosis has a physiologically important role in regulating denervation-induced muscle atrophy.

MeSH Terms
Animals Apoptosis/physiology Gene Expression Regulation/physiology Mitochondria, Muscle/metabolism Muscle Denervation/methods Muscle Proteins/metabolism Muscle, Skeletal/innervation,metabolism Rats Rats, Inbred F344 Signal Transduction/physiology
Chemicals
Muscle Proteins
Authors & Affiliations
2 authors, click to expand affiliations / ORCID
Siu Parco M
Laboratory of Muscle Biology an Sarcopenia, Division of Exercise Physiology, West Virginia University School of Medicine, Morgantown WV 26506-9227, USA.
Alway Stephen E
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Article Info
Journal
The Journal of physiology
Abbr.
J Physiol
ISSN
0022-3751
DOI
10.1113/jphysiol.2004.081083
Published
2005-05-15
Epub
2005-00-17
Pages
309-23
Language
English
Region
England
NLM ID
0266262
PMCID
PMC1464486
Subset
IM
Grants
NIA NIH HHS · R01 AG021530 · United States
External Links
PubMed source DOI: 10.1113/jphysiol.2004.081083 PMC Full Text View journal details
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