The aim of this experiment is to study mitochondrial functional status in Arabidopsis embryo sacs using the membrane potential indicator JC-1. Changes in the membrane potential are presumed to be due to the opening of the mitochondrial permeability transition pore (MPTP), allowing passage of ions and small molecules. The resulting equilibrium of ions leads in turn to the decoupling of the respiratory chain and the release of cytochrome c into the cytosol, a distinctive feature of the early stages of programmed cell death.
JC-1 is a lipophilic dye that can selectively enter into mitochondria and reversibly change color from green to red as the membrane potential increases. In healthy cells with high mitochondrial potential, JC-1 spontaneously forms complexes with intense red fluorescence. On the other hand, in mitochondria with low mitochondrial potential, JC-1 remains in the monomeric form, which exhibits only green fluorescence (Martin et al., 2013; Hauser et al., 2006).
This protocol could be used in isolated mitochondria, and in a variety of cell types and different tissues of plants and other organism.
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