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© Research
Publication : Experimental cell research

Real-time flow cytometry analysis of permeability transition in isolated mitochondria

Scientific Fields
Diseases
Organisms
Applications
Technique

Published in Experimental cell research - 10 Mar 2004

Lecoeur H, Langonné A, Baux L, Rebouillat D, Rustin P, Prévost MC, Brenner C, Edelman L, Jacotot E

Link to Pubmed [PMID] – 14980506

Exp. Cell Res. 2004 Mar;294(1):106-17

Mitochondrial membrane permeabilization (MMP) is a key event in necrotic and (intrinsic) apoptotic processes. MMP is controlled by a few major rate-limiting events, one of which is opening of the permeability transition pore (PTP). Here we develop a flow cytometry (FC)-based approach to screen and study inducers and blockers of MMP in isolated mitochondria. Fixed-time and real-time FC permits to co-evaluate and order modifications of mitochondrial size, structure and inner membrane (IM) electrochemical potential (DeltaPsi(m)) during MMP. Calcium, a major PTP opener, and alamethicin, a PTP-independent MMP inducer, trigger significant mitochondrial forward scatter (FSC) increase and side scatter (SSC) decrease, correlating with spectrophotometrically detected swelling. FC-based fluorescence detection of the DeltaPsi(m)-sensitive cationic lipophilic dye JC-1 permits to detect DeltaPsi(m) variations induced by PTP openers or specific inducers of inner MMP such as carbonylcyanide m-chlorophenylhydrazone (mClCCP). These simple, highly sensitive and quantitative FC-based methods will be pertinent to evaluate compounds for their ability to control MMP.

http://www.ncbi.nlm.nih.gov/pubmed/14980506