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Updated: Apr 20, 2026
Analyzing Mitochondrial Transport and Morphology in Human Induced Pluripotent Stem Cell-Derived Neurons in Hereditary Spastic Paraplegia
Published on: February 9, 2020
Monitoring mitochondrial membranes permeability in live neurons and mitochondrial swelling through electron
Macarena S Arrázola1, Nibaldo C Inestrosa
1Departamento de Biología Celular y Molecular, Facultad de Ciencias Biológicas, Centro de Envejecimiento y Regeneración (CARE), Pontificia Universidad Católica de Chile, Santiago, Chile.
Abstract:
Maintenance of mitochondrial membrane integrity is essential for mitochondrial function and neuronal viability. Apoptotic stimulus or calcium overload leads to mitochondrial permeability transition pore (mPTP ) opening and induces mitochondrial swelling, a common feature of mitochondrial membrane permeabilization. The first phenomenon can be evaluated in cells loaded with the dye calcein -AM quenched by cobalt, and mitochondrial swelling can be detected by electron microscopy through the analysis of mitochondrial membrane integrity. Here, we describe a live cell imaging assay to detect mitochondrial permeability transition and the development of a detailed analysis of morphological and ultrastructural changes that mitochondria undergo during this process.
Insights
This study introduces a live cell imaging assay to detect mitochondrial permeability transition pore opening and swelling. This method aids in understanding mitochondrial dysfunction and its impact on neuronal viability.
Area of Science:
- Cell Biology
- Neuroscience
- Mitochondrial Biology
Background:
- Mitochondrial membrane integrity is crucial for cellular function and survival, particularly in neurons.
- Mitochondrial permeability transition pore (mPTP) opening, triggered by apoptosis or calcium overload, causes mitochondrial swelling and membrane permeabilization.
Purpose of the Study:
- To describe a novel live cell imaging assay for detecting mitochondrial permeability transition.
- To develop a detailed analysis of mitochondrial morphological and ultrastructural changes during this process.
Main Methods:
- Utilizing calcein-AM dye quenched by cobalt to evaluate mPTP opening in live cells.
- Employing electron microscopy to analyze mitochondrial swelling and membrane integrity.
Main Results:
- Successfully established a live cell imaging assay to monitor mitochondrial permeability transition.
- Characterized morphological and ultrastructural alterations in mitochondria during mPTP opening.
Conclusions:
- The developed assay provides a valuable tool for studying mitochondrial dysfunction.
- This research enhances the understanding of mitochondrial membrane permeabilization and its consequences.
