Photo-induction and automated quantification of reversible mitochondrial permeability transition pore opening in primary mouse myotubes
Publication year
2014Source
PLoS One, 9, 11, (2014), article e114090ISSN
Publication type
Article / Letter to editor
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Organization
Biochemistry (UMC)
Paediatrics - OUD tm 2017
Journal title
PLoS One
Volume
vol. 9
Issue
iss. 11
Subject
Radboudumc 6: Metabolic Disorders RIMLS: Radboud Institute for Molecular Life SciencesAbstract
Opening of the mitochondrial permeability transition pore (mPTP) is involved in various cellular processes including apoptosis induction. Two distinct states of mPTP opening have been identified allowing the transfer of molecules with a molecular weight <1500 Da or <300 Da. The latter state is considered to be reversible and suggested to play a role in normal cell physiology. Here we present a strategy combining live-cell imaging and computer-assisted image processing allowing spatial visualization and quantitative analysis of reversible mPTP openings ("DeltaPsi flickering") in primary mouse myotubes. The latter were stained with the photosensitive cation TMRM, which partitions between the cytosol and mitochondrial matrix as a function of mitochondrial membrane potential (DeltaPsi). Controlled illumination of TMRM-stained primary mouse myotubes induced DeltaPsi flickering in particular parts of the cell ("flickering domains"). A novel quantitative automated analysis was developed and validated to detect and quantify the frequency, size, and location of individual DeltaPsi flickering events in myotubes.
This item appears in the following Collection(s)
- Academic publications [245050]
- Electronic publications [132309]
- Faculty of Medical Sciences [93209]
- Open Access publications [105922]
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