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Apoptosis in differentiating C2C12 muscle cells selectively targets Bcl-2-deficient myotubes

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Abstract

Muscle cell apoptosis accompanies normal muscle development and regeneration, as well as degenerative diseases and aging. C2C12 murine myoblast cells represent a common model to study muscle differentiation. Though it was already shown that myogenic differentiation of C2C12 cells is accompanied by enhanced apoptosis in a fraction of cells, either the cell population sensitive to apoptosis or regulatory mechanisms for the apoptotic response are unclear so far. In the current study we characterize apoptotic phenotypes of different types of C2C12 cells at all stages of differentiation, and report here that myotubes of differentiated C2C12 cells with low levels of anti-apoptotic Bcl-2 expression are particularly vulnerable to apoptosis even though they are displaying low levels of pro-apoptotic proteins Bax, Bak and Bad. In contrast, reserve cells exhibit higher levels of Bcl-2 and high resistance to apoptosis. The transfection of proliferating myoblasts with Bcl-2 prior to differentiation did not protect against spontaneous apoptosis accompanying differentiation of C2C12 cells but led to Bcl-2 overexpression in myotubes and to significant protection from apoptotic cell loss caused by exposure to hydrogen peroxide. Overall, our data advocate for a Bcl-2-dependent mechanism of apoptosis in differentiated muscle cells. However, downstream processes for spontaneous and hydrogen peroxide induced apoptosis are not completely similar. Apoptosis in differentiating myoblasts and myotubes is regulated not through interaction of Bcl-2 with pro-apoptotic Bcl-2 family proteins such as Bax, Bak, and Bad.

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Abbreviations

DM:

Differentiation medium

ER:

Endoplasmic reticulum

GM:

Growing medium

TG:

Thapsigargin

TIC:

Total ion current

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Acknowledgments

This study was supported by the NIH (P01AG012993). The authors gratefully acknowledge Drs. J. Enouf and R. Bobe (Inserm, Hopital Lariboisiere, Paris, France) for the gift of anti-SERCA2B antibodies.

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Authors and Affiliations

  1. University of Kansas, Lawrence, KS, USA

    Christian Schöneich

  2. Department of Pharmaceutical Chemistry, University of Kansas, Lawrence, KS, 66047, USA

    Elena Dremina & Victor Sharov

  3. Mass Spectrometry Laboratory, University of Kansas, Lawrence, KS, 66047, USA

    Nadezhda Galeva

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  1. Christian Schöneich
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  2. Elena Dremina
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  3. Nadezhda Galeva
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  4. Victor Sharov
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Correspondence to Christian Schöneich.

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Schöneich, C., Dremina, E., Galeva, N. et al. Apoptosis in differentiating C2C12 muscle cells selectively targets Bcl-2-deficient myotubes. Apoptosis 19, 42–57 (2014). https://doi.org/10.1007/s10495-013-0922-7

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