A method for measuring mitochondrial mass and activity

Abstract

Mitochondria, responsible for the energygenerating process essential for the cell metabolism, differ for the number, localization and activity in animal cells and tissues in relation to the energetic needs. Using fluorescent probes specific for mitochondria, Mitotracker Green (MTG) and Orange (MTO), and Confocal Laser-Scanning Microscope (CLSM), we elaborated a method to measure in vivo the mitochondrial mass and activity, in sea urchin Paracentrotus lividus eggs and embryos. The analysis of captured images, revealed a variation of mitochondrial distribution and an increase of activity after fertilization.

Hydrogenosome Metabolism Is the Key Target for Antiparasitic Activity of Resveratrol against Trichomonas vaginalis

“…MitoTracker Deep Red FM is a far-red-fluorescent dye (excitation/emission ϳ644/665 nm) that stains mitochondria and hydrogenosomes in live cells and can be used for in vivo determination of the mass and localization of the hydrogenosome in T. vaginalis (61,62). We noted that both RESV and MDZ significantly increased the fluorescence emitted by the parasite hydrogenosomes incubated with MitoTracker Deep Red FM, suggesting that both compounds induce changes in hydrogenosomal metabolism in T. vaginalis trophozoites.…”

2-Methoxyestradiol Affects Mitochondrial Biogenesis Pathway and Succinate Dehydrogenase Complex Flavoprotein Subunit A in Osteosarcoma Cancer Cells

“…After 24 h, cells were incubated with 2-ME and/or L-lactate according to the experimental design. Thirty minutes before the end of incubation, MitoTracker ® Green FM (200 nM) was added to the culture medium (31 with 2-ME and/or L-lactate according to the experimental design and fixed with 4% paraformaldehyde for 15 min. Following antigen retrieval and washing with PBS, cells were permeabilized with 0.1% Triton X-100 in PBS for 5 min and blocked in 10x blocking solution for 1 h at room temperature.…”

Viologen-phosphorus dendrimers exhibit minor toxicity against a murine neuroblastoma cell line

“…CM-H 2 TMRos is oxidized inside the cell to the fluorescent CMTMRos and then selectively sequestered in the mitochondria [35]. Cells were seeded in 12-well plates at a density of 25 × 10 4 cells/well in DMEM medium and incubated for 20 h. After 24 h dendrimer treatment, the supernatant was discarded, and the cells were washed with PBS and stained with 500 nM CM-H 2 TMRos for 30 min in growing conditions.…”