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Isolation and Differentiation of Primary Myoblasts from Mouse Skeletal Muscle Explants
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Differentiation activates mitochondrial OPA1 processing in myoblast cell lines.

Harpreet Kaur1, Omar Carrillo1, Iraselia Garcia2

  • 1School of Integrative Biological & Chemical Sciences, The University of Texas Rio Grande Valley, United States.

Mitochondrion
|July 10, 2024
PubMed
Summary
This summary is machine-generated.

Mitochondrial OPA1 processing becomes sensitive to membrane potential loss during myoblast differentiation, regulated by mitochondrial stress. This process impacts apoptosis and differentiation.

Keywords:
DifferentiationMitochondriaOMA1OPA1Transmembrane potential

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Area of Science:

  • Mitochondrial biology
  • Cellular stress response
  • Developmental biology

Background:

  • Mitochondrial optic atrophy-1 (OPA1) regulates mitochondrial structure and function.
  • OPA1 processing is linked to inner membrane potential (Δψm) and cellular stress.
  • OPA1 is typically cleaved to inactive forms upon Δψm loss, but this was previously found to be insensitive in undifferentiated H9c2 cells.

Purpose of the Study:

  • To investigate the developmental regulation of OPA1 processing in myoblasts.
  • To identify mechanisms controlling OPA1 sensitivity to Δψm loss during differentiation.
  • To explore the role of OPA1 in myoblast differentiation and apoptosis.

Main Methods:

  • Utilized H9c2 cardiomyoblasts, L6.C11, and C2C12 myoblast cell lines.
  • Induced differentiation using low serum media.
  • Administered carbonyl cyanide chlorophenyl hydrazone (CCCP) to disrupt Δψm and chloramphenicol (CAP) to inhibit mitochondrial protein synthesis.
  • Performed OPA1 knockdown experiments.

Main Results:

  • OPA1 processing became sensitive to Δψm loss upon differentiation, independent of ATRA.
  • Chloramphenicol (CAP) pretreatment mimicked differentiation-induced OPA1 processing sensitivity.
  • This regulation was observed across multiple myoblast lines, suggesting a general mechanism.
  • Restored OPA1 processing correlated with increased apoptosis sensitivity.
  • OPA1 is essential for effective myoblast differentiation.

Conclusions:

  • A novel developmental mechanism regulates OMA1-mediated OPA1 processing in myoblasts.
  • Myoblast differentiation engages mitochondrial stress sensing pathways that alter OPA1 processing.
  • OPA1 processing dynamics are critical for balancing mitochondrial integrity, differentiation, and apoptosis.