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Functional, morphological, and apoptotic alterations in skeletal muscle of ARC deficient mice

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Abstract

Apoptotic signaling plays an important role in the development and maintenance of healthy skeletal muscle. However, dysregulation of apoptotic signals in skeletal muscle is associated with atrophy and loss of function. Apoptosis repressor with caspase recruitment domain (ARC) is a potent anti-apoptotic protein that is highly expressed in skeletal muscle; however, its role in this tissue has yet to be elucidated. To investigate whether ARC deficiency has morphological, functional, and apoptotic consequences, skeletal muscle from 18 week-old wild-type and ARC knockout (KO) mice was studied. In red muscle (soleus), we found lower maximum tetanic force, as well as a shift towards a greater proportion of type II fibers in ARC KO mice. Furthermore, the soleus of ARC KO mice exhibited lower total, as well as fiber type-specific cross sectional area in type I and IIA fibers. Interestingly, these changes in ARC KO mice corresponded with increased DNA fragmentation, albeit independent of caspase or calpain activation. However, cytosolic fractions of red muscle from ARC KO mice had higher apoptosis inducing factor content, suggesting increased mitochondrial-mediated, caspase-independent apoptotic signaling. This was confirmed in isolated mitochondrial preparations, as mitochondria from skeletal muscle of ARC KO mice were more susceptible to calcium stress. Interestingly, white muscle from ARC KO mice showed no signs of altered apoptotic signaling or detrimental morphological differences. Results from this study suggest that even under basal conditions ARC influences muscle apoptotic signaling, phenotype, and function, particularly in slow and/or oxidative muscle.

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Abbreviations

½RT:

Half relaxation time

AIF:

Apoptosis inducing factor

ANT:

Adenine nucleotide translocator

ARC:

Apoptosis repressor with caspase recruitment domain

AU:

Arbitrary units

Bax:

Bcl-2 associated X protein

Bcl-2:

B cell lymphoma 2

CLAMS:

Comprehensive Lab Animal Monitoring System

CSA:

Cross sectional area

CuZnSOD:

Copper zinc superoxide dismutase

Cyto C:

Cytochrome C

EndoG:

Endonuclease G

ER:

Endoplasmic reticulum

FLIP:

FLICE-like inhibitory protein

GPD:

α-Glycerophosphate dehydrogenase

HET:

Heterozygous

HMW:

High molecular weight

Hsp70:

Heat shock protein 70

KO:

Knockout

LC3B:

Microtubule-associated protein 1 light chain 3 beta

MAFbx:

Muscle atrophy F-box

MOMP:

Mitochondrial outer membrane permeabilization

MuRF1:

Muscle RING-finger protein-1

p62:

p62/sequestosome 1

PFK:

Phosphofructokinase

PTP:

Permeability transition pore

R123:

Rhodamine 123

RQ:

Red quadriceps

SDH:

Succinate dehydrogenase

Smac:

Second mitochondria-derived activator of caspase

TPT:

Time to peak tension

TUNEL:

Terminal deoxynucleotidyl transferase dUTP nick end labeling

UPS:

Ubiquitin proteasome system

\({\dot{\hbox{V}}}{\hbox{O}}_2\) :

Oxygen consumption

WQ:

White quadriceps

WT:

Wild-type

XIAP:

X-linked inhibitor of apoptosis

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Acknowledgments

This research was supported by funds provided to Joe Quadrilatero by the Natural Sciences and Engineering Research Council of Canada (NSERC) and the Canadian Institutes of Health Research (CIHR). The authors declare no competing financial interests.

Conflict of interest

None declared.

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Authors and Affiliations

  1. Department of Kinesiology, University of Waterloo, Waterloo, ON, N2L3G1, Canada

    Andrew S. Mitchell, Ian C. Smith, Daniel Gamu, A. Russell Tupling & Joe Quadrilatero

  2. Max-Delbrück Center for Molecular Medicine, Berlin, Germany

    Stefan Donath

  3. Center for Stroke Research Berlin, Charite-University Medicine, Berlin, Germany

    Stefan Donath

  4. Department of Cardiology and Nephrology, HELIOS Clinics GmbH, Berlin, Germany

    Stefan Donath

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  1. Andrew S. Mitchell
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  2. Ian C. Smith
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Correspondence to Joe Quadrilatero.

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Mitchell, A.S., Smith, I.C., Gamu, D. et al. Functional, morphological, and apoptotic alterations in skeletal muscle of ARC deficient mice. Apoptosis 20, 310–326 (2015). https://doi.org/10.1007/s10495-014-1078-9

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