What does JC-1 reveal about mitochondria?

The membrane-permeant JC-1 dye is widely used in apoptosis studies to monitor mitochondrial health. JC-1 dye can be used as an indicator of mitochondrial membrane potential in a variety of cell types, including myocytes and neurons, as well as in intact tissues and isolated mitochondria.

What is mitochondrial membrane potential?

The mitochondrial membrane potential (ΔΨm) generated by proton pumps (Complexes I, III and IV) is an essential component in the process of energy storage during oxidative phosphorylation. Together with the proton gradient (ΔpH), ΔΨm forms the transmembrane potential of hydrogen ions which is harnessed to make ATP.

What is the mitochondrial membrane potential?

What is CCCP treatment?

The CCCP treatment was found to significantly reduce ATP content, increase the amount of phosphorylated AMP-activated protein kinase and elevate reactive oxygen species levels in oocytes.

What does a positive membrane potential mean?

For a cell’s membrane potential, the reference point is the outside of the cell. Because there is a potential difference across the cell membrane, the membrane is said to be polarized. If the membrane potential becomes more positive than it is at the resting potential, the membrane is said to be depolarized.

What is resting mitochondrial membrane potential?

Mitochondrial membrane potential (ΔΨM) is a central intermediate in oxidative energy metabolism. In cultured rat cortical neurons, ΔΨM is −139 mV at rest, and is regulated between −108 mV and −158 mV by concerted increases in ATP demand and Ca2+-dependent metabolic activation.

What is the function of membrane potential?

The membrane potential represents a balance among the equilibrium potentials of the ions to which the membrane is permeable. The greater the conductance of an ion, the more that ion will influence the membrane potential of the cell.

What is the difference between FCCP and CCCP?

FCCP applied for 30 minutes at 10 microM reduces the number of moving organelles by 78% and a similar treatment with CCCP causes a reduction of 47%. At 100 microM either compound abolishes all directed movements both in neurites and in cultured 3T3 cells.

What is membrane potential dye used for?

FluoVolt Membrane Potential Dye can be used for imaging electrical activity from intact heart tissues, mapping membrane potentials along neurons and muscle fibers, or measuring potential changes in response to pharmacological stimuli. ANEP dyes fluoresce in response to electrical potential changes in their environment.

Can JC-1 dye be used as a quantitative measure of membrane potential?

It is noteworthy JC-1 dye can be used both as qualitative (considering the shift from green to red fluorescence emission) and quantitative (considering only the pure fluorescence intensity) measure of mitochondrial membrane potential (Cossarizza and Salvioli, 1998).

How reliable are fluorescent dyes for monitoring mitochondrial membrane potential?

In recent years, fluorescent dyes have been frequently used for monitoring mitochondrial membrane potential to evaluate mitochondrial viability and function. However, the reproducibility of the results across laboratories strongly depends upon following well validated and reliable protocols along with the appropriate controls.

Can fluovolt membrane potential dye detect fluorogenic responses in human HEK 293 cells?

Human HEK 293 cells were loaded with FluoVolt Membrane Potential Dye. Cells were imaged with 10 millisecond illumination pulses and images acquired with 2X binning. Traces show fluorogenic responses as cells are depolarized (A) at 2 second intervals from –100 mV to +30 mV or (B) in single steps from –80 mV to 0 mV at 2 second intervals.