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Updated: Jul 7, 2026
Understanding the Changes in Mitochondrial Morphology through Dynamic and Three-dimensional Fluorescence Micrographs
Published on: August 15, 2025
Measuring mitochondrial shape changes and their consequences on mitochondrial involvement during apoptosis
Christian Frezza1, Sara Cipolat, Luca Scorrano
1Dulbecco-Telethon Institute, Venetian Institute of Molecular Medicine, Padova, Italy.
Abstract:
Mitochondria are key players in cell death following intrinsic and, in some cell types, extrinsic stimuli. The recruitment of the mitochondrial pathway results in mitochondrial dysfunction and release of intermembrane space proteins like cytochrome-c that are required in the cytosol for complete activation of effector caspases. Apoptotic shape changes of this organelle and the role of "mitochondria-shaping" proteins in cell death has attracted considerable attention. We present protocols to investigate how morphological changes of the mitochondrial reticulum regulate release of cytochrome-c, as evaluated quantitatively by an in situ approach, and changes in mitochondrial membrane potential measured in real time.
Insights
Mitochondria
Area of Science:
- Cell Biology
- Biochemistry
Background:
- Mitochondria are central to intrinsic and extrinsic cell death pathways.
- Mitochondrial dysfunction leads to the release of cytochrome-c, activating caspases.
- The role of mitochondria-shaping proteins in apoptosis is a key research area.
Purpose of the Study:
- To investigate how mitochondrial reticulum morphology influences cytochrome-c release.
- To quantify cytochrome-c release using an in situ approach.
- To measure real-time changes in mitochondrial membrane potential.
Main Methods:
- Quantitative in situ analysis of cytochrome-c release.
- Real-time measurement of mitochondrial membrane potential.
- Investigation of mitochondria-shaping proteins' role in apoptosis.
Main Results:
- Protocols were developed to study mitochondrial morphology and cell death.
- Morphological changes in the mitochondrial reticulum were linked to cytochrome-c release.
- Mitochondrial membrane potential changes were monitored in real time.
Conclusions:
- Morphological dynamics of the mitochondrial reticulum are critical regulators of apoptosis.
- The presented protocols enable quantitative assessment of mitochondrial involvement in cell death.
- Understanding these mechanisms can inform therapeutic strategies for diseases involving cell death.
