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Isolation and Functional Analysis of Mitochondria from Cultured Cells and Mouse Tissue
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Isolation and Functional Analysis of Mitochondria from Cultured Cells and Mouse Tissue

Published on: March 23, 2015

Apoptosis-associated mitochondrial outer membrane permeabilization assays

Damien Arnoult1

  • 1INSERM U 542, Université Paris-Sud, Hopital Paul Brousse, Batiment Lavoisier, 14 avenue Paul Vaillant Couturier, 94807 Villejuif cedex, France. damarn@vjf.inserm.fr

Methods (San Diego, Calif.)
|March 4, 2008
PubMed

Insights

This study details methods for measuring cytochrome c release from cells and isolated mitochondria to assess apoptosis. Assays for Bax/Bak activation and oligomerization are also presented, aiding cell death research.

Area of Science:

  • Biochemistry
  • Cell Biology
  • Molecular Biology

Background:

  • Apoptosis involves the activation of pro-apoptotic proteins like Bax and Bak.
  • Activated Bax and Bak oligomerize, permeabilizing the mitochondrial outer membrane.
  • This permeabilization releases cytochrome c, initiating caspase activation and cell death.

Purpose of the Study:

  • To provide detailed protocols for measuring cytochrome c release.
  • To enable assessment of Bax/Bak activation and mitochondrial outer membrane permeabilization.
  • To offer methods for studying apoptosis in both intact cells and isolated mitochondria.

Main Methods:

  • Quantification of cytochrome c release from intact cultured cells.
  • Measurement of cytochrome c release from isolated functional mitochondria.
  • Assays to detect Bax and Bak activation and oligomerization.

Main Results:

  • Established protocols for assessing key apoptosis events.
  • Demonstrated the utility of both cellular and isolated mitochondrial assays.
  • Provided tools for investigating the mechanisms of mitochondrial outer membrane permeabilization.

Conclusions:

  • Cytochrome c release is a critical indicator of apoptosis and mitochondrial outer membrane permeabilization.
  • The presented methods facilitate robust investigation of apoptosis.
  • These protocols support research into cell death pathways and therapeutic interventions.

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