| Literature DB >> 14980506 |
Hervé Lecoeur1, Alain Langonné, Ludwig Baux, Dominique Rebouillat, Pierre Rustin, Marie-Christine Prévost, Catherine Brenner, Léna Edelman, Etienne Jacotot.
Abstract
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.Entities:
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Year: 2004 PMID： 14980506 DOI： 10.1016/j.yexcr.2003.10.030
Source DB: PubMed Journal: Exp Cell Res ISSN： 0014-4827 Impact factor: 3.905