Mitochondrial Permeabilization
Mitochondrial Permeabilization - Related Videos
05:47
Oxygen Flux Measurements to Evaluate Mitochondrial Respiration in Living and Permeabilized Cells
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2026
Source: Elettra Leo1, Luiza H.D. Cardoso1, Sabine Schmitt1, Erich Gnaiger11Oroboros InstrumentsThis video explains how oxygen flux is used to assess mitochondrial respiration in living and permeabilized mammalian cells. It demonstrates the sequential use of digitonin, metabolic substrates, ADP, an uncoupler, and an inhibitor to characterize different respiratory states. The resulting oxygen flux profile reveals mitochondrial function and energy-coupling efficiency under defined conditions.
08:33
High-resolution Respirometry to Assess Mitochondrial Function in Permeabilized and Intact Cells
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Cited by 62 •
2017
High-resolution respirometry is used to determine mitochondrial oxygen consumption. This is a straightforward technique to determine mitochondrial respiratory chain complexes' (I-IV) respiratory rates, maximal mitochondrial electron transport system capacity, and mitochondrial outer membrane integrity.
06:17
Measuring Mitochondrial Substrate Flux in Recombinant Perfringolysin O-Permeabilized Cells
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Cited by 1 •
2021
In this work, we describe a modified protocol to test mitochondrial respiratory substrate flux using recombinant perfringolysin O in combination with microplate-based respirometry. With this protocol, we show how metformin affects mitochondrial respiration of two different tumor cell lines.
14:55
Measurement of Mitochondrial Oxygen Consumption in Permeabilized Fibers of Drosophila Using Minimal Amounts of Tissue
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Cited by 31 •
2018
In this paper, a method to measure oxygen consumption using high-resolution respirometry in permeabilized thoraxes of Drosophila is described. This technique requires a minimal amount of tissue compared to the classic mitochondrial isolation technique and the results obtained are more physiologically relevant.
08:43
Simultaneous Measurement of Mitochondrial Calcium and Mitochondrial Membrane Potential in Live Cells by Fluorescent Microscopy
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Cited by 29 •
2017
Mitochondria can utilize the electrochemical potential across their inner membrane (ΔΨm) to sequester calcium (Ca2+), allowing them to shape cytosolic Ca2+ signaling within the cell. We describe a method for simultaneously measuring mitochondria Ca2+ uptake and ΔΨm in live cells using fluorescent dyes and confocal microscopy.
FAQs
During apoptosis, the pro-apoptotic proteins BAX and BAK form pores in the mitochondrial outer membrane. These openings allow cytochrome c and other proteins from the intermembrane space to escape into the cell, providing a molecular connection between mitochondrial damage and downstream apoptotic signaling. Their pore-forming activity is therefore central to mitochondrial control of programmed cell death.
Outer-membrane pore formation releases cytochrome c and other intermembrane-space proteins, whereas disruption associated with the inner membrane can collapse the mitochondrial membrane potential. Calcium overload, oxidative stress, or permeability transition may contribute to this inner-membrane failure. Loss of the potential impairs ATP production, linking mitochondrial permeabilization to both cell-death signaling and energy failure.
The affected membrane and initiating stress influence the resulting cellular outcome. Outer-membrane permeabilization can release proteins that support apoptosis, while inner-membrane disruption can reduce membrane potential and ATP production. Because mitochondrial stress can connect with apoptosis, necrosis, or altered energy metabolism, distinguishing these mechanisms helps explain why different forms of damage produce different consequences.
Several stress conditions can compromise mitochondrial barrier function, including calcium overload, oxidative stress, and permeability transition. These factors may affect the inner membrane, causing membrane-potential collapse and impaired ATP production, while apoptotic signaling can activate BAX and BAK at the outer membrane. Identifying the initiating stress helps researchers connect mitochondrial changes with the observed cell-death pathway.
Studies can focus on whether BAX and BAK form outer-membrane pores, whether cytochrome c or other intermembrane-space proteins are released, and whether the inner-membrane potential is lost. Researchers can also consider consequences for ATP production and cell fate. Examining these linked outcomes helps distinguish apoptotic signaling from broader mitochondrial dysfunction and energy failure.
Mitochondrial permeabilization provides a framework for studying how cellular stress contributes to neurodegeneration and how altered cell-death control supports cancer biology. In these contexts, researchers can investigate mitochondrial dysfunction, apoptotic signaling, and energy disruption as connected processes. The pathway therefore helps relate molecular membrane changes to disease-associated cell survival or loss.
Therapeutic compounds may be evaluated for their ability to influence mitochondrial pathways that control cell survival. Understanding permeabilization can support strategies designed to prevent unwanted cell loss or selectively trigger death in diseased cells. Researchers can assess effects on protein release, membrane potential, ATP production, and downstream cell fate to connect compound activity with mitochondrial mechanisms.
Because mitochondrial barrier failure can contribute to apoptosis, necrosis, and impaired energy metabolism, understanding its triggers may reveal opportunities to limit unwanted damage. Research can focus on preventing stress-associated disruption, preserving membrane potential, or maintaining ATP production. Such information is relevant when the goal is to protect cells from pathological mitochondrial dysfunction rather than promote diseased-cell death.