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High Zn2+ in the mitochondria induces mitochondrial membrane potential collapse, which activates caspase-3 and leads to cell apoptosis and lactate dehydrogenase (LDH) release [15, 16].
The ultimate effect of essential oils is cell death due to permeabilization of the cellular membrane, loss of ions, reduction of membrane potential, collapse of the proton pump and depletion of the ATP pool (Bakkali et al. 2008).
Mitochondrial membrane potential collapse marks a point-of-no-return during apoptosis and occurs earlier than DNA fragmentation [38], [39].
FCCP, a mitochondrial uncoupler, was used as control because it induced a mitochondrial membrane potential collapse in intact cells.
Analysis of mitochondrial membrane potential collapse by fluorescence microscopy using MitoLight dye indicated that pU, pC and pCU caused significant loss of mitochondrial membrane potential (Δψm) as indicated by a decrease in red fluorescence as compared to controls (Fig. 1H).
However, events other than mitochondrial Ca2+ uptake are required for mitochondrial membrane potential collapse.
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In apoptotic cells, the mitochondrial membrane potential collapses, and JC-1 cannot accumulate within the mitochondria.
We used the dye DiSC3 5) to probe the inner membrane potential because the fluorescence of the dye increases when the membrane potential collapses.
As soon as the mitochondrial membrane potential collapses, JC-1 is dispersed throughout the cell in a monomeric form (green fluorescence).
When the mitochondrial membrane potential collapses, the JC-1 cannot accumulate within the mitochondria and exists as monomers in the cytosol, which are detected as green fluorescence light (R2 as shown in Figure 2(a)).
The resulting Ca2+ accumulation has been shown to contribute to mitochondrial membrane-potential collapse (Vergun et al., 1999).
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