Rapid and quantitative in vitro analysis of mitochondrial fusion and its interplay with apoptosis

摘要: Mitochondrial fusion is essential to maintain genomic stability and physiological functions of mitochondria. Since mitochondrial fission work in concert regulate morphology functions, it has been challenging quantitatively measure the direct roles apoptosis cancer progression. Here, we report development a high-throughput vitro method quantify through single mitochondria analysis by laboratory-built nano-flow cytometer (nFCM). Isolated expressing green fluorescent protein (GFP-mito) or discosoma red (DsRed-mito) were mixed together, induced fuse, analyzed nFCM. A particle exhibiting both fluorescence was identified as an event heterotypic fusion, efficiency used surrogate overall efficiency. The as-developed applied reveal interplay between without interference fission. We show that cytosolic components promoted this upregulation diminished during apoptosis. Combined with translocation Bid Bax from cytosol mitochondria, these findings suggest pro-apoptotic Bcl-2 family proteins could be positive mediators fusion. On other hand, also renders more resistant membrane potential collapse upon induction. Our data disruption potent strategy for therapy. Furthermore, offers effective approach identify inhibitors, including betulinic acid antimycin A, giving reasons their powerful utility treatment.