[Flow cytometric analysis for detecting mitochondrial permeability transition pore opening].
OBJECTIVE
To introduce a new method for detecting mitochondrial permeability transition pore (PTP) opening with flow cytometry using the resveratrol-inducing PTP opening model.


METHODS
Mitochondria were isolated from rat livers and selectively labeled with nonyl acridine orange. The mitochondrial membrane potential was detected using flow cytometry with TMRE (tetramethylrhodamine, ethyl ester) labeling. PTP opening induced by resveratrol was represented by the changes of mitochondrial side-scattering (SSC) detected by flow cytometry.


RESULTS
Flow cytometry was capable of defining the purity of the mitochondria isolated. The fluorescence intensities and SSC of the mitochondria were decreased after resveratrol treatment, indicating that resveratrol could induce PTP opening. Ciclosporin A inhibited resveratrol-induced PTP opening.


CONCLUSIONS
Flow cytometric analysis allows accurate and convenient detection of mitochondrial membrane potential, mitochondrial swelling and PTP opening.
Yan
Fang
F
Department of Histology and Embryology, Southern Medical University, Guangzhou 510515, China. yanfang@fimmu.com
Ma
Xiao-dong
XD
Tian
Xue-mei
XM
D000818:Animals; D017209:Apoptosis; D005434:Flow Cytometry; D053078:Membrane Potential, Mitochondrial / Q000235:genetics*; D008930:Mitochondria, Liver / Q000378:metabolism; D033681:Mitochondrial Membrane Transport Proteins / Q000378:metabolism*; D000083162:Mitochondrial Permeability Transition Pore; D051381:Rats; D012235:Rhodamines
D033681:Mitochondrial Membrane Transport Proteins; D000083162:Mitochondrial Permeability Transition Pore; D012235:Rhodamines; C005358:tetramethylrhodamine