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The Intrinsic Apoptotic Pathway

The Intrinsic Apoptotic Pathway

Internal cellular stress, such as cellular injury or hypoxia, triggers intrinsic apoptosis. The B-cell lymphoma 2 (Bcl-2) family of proteins are the primary regulators of the intrinsic apoptotic pathway. For example, during DNA damage, checkpoint proteins, such as Ataxia Telangiectasia Mutated (ATM protein) and Checkpoints Factor-2 (Chk2) proteins, are activated. These proteins phosphorylate p53 which further activates pro-apoptotic proteins, such as Bax, Bak, PUMA, and Noxa, and inhibits...

Apoptosis

Apoptosis

Apoptosis is a combination of two Greek words, 'apo' and 'ptosis,' meaning separation and falling off, respectively. Hippocrates used this word to describe gangrene, which was caused due to bandaging of fractured bones. Apoptosis was distinguished from necrosis in 1970 when John Kerr reported observations of morphological changes occurring during apoptosis. During one experiment, he observed that the disruption of blood supply to the liver tissue resulted in a size reduction of the tissue.

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Related Experiment Video

Updated: Jul 7, 2026

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

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

Summary

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.

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Last Updated: Jul 7, 2026

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

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Published on: March 23, 2015

Detection of Mitochondria Membrane Potential to Study CLIC4 Knockdown-induced HN4 Cell Apoptosis In Vitro

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Use of a Caspase Multiplexing Assay to Determine Apoptosis in a Hypothalamic Cell Model

Use of a Caspase Multiplexing Assay to Determine Apoptosis in a Hypothalamic Cell Model

Published on: April 16, 2014

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.