title: Involvement of cyclophilin D in mitochondrial permeability transition induction in intact cells.
abstract: The mitochondrial permeability transition (MPT) is involved in both Ca(2+) signaling and cell death. The present study aimed to clarify the involvement of cyclophilin D, a peptidyl prolyl cis-trans isomerase (PPIase), in MPT induction in intact cells. To achieve this, we used C6 cells overexpressing wild-type or PPIase-deficient cyclophilin D, and measured the inner mitochondrial membrane permeability to calcein, a 623-Da hydrophilic fluorescent molecule, to evaluate MPT induction. In vector control cells, the percentage of MPT induction by ionomycin increased as the Ca(2+) concentration in the extracellular medium increased. This result indicates that the present method is valid for numerical evaluation of MPT induction. In C6 cells expressing the PPIase-deficient mutant, the percentage of MPT induction was significantly decreased compared with wild-type CypD-overexpressing cells or vector control cells. These results suggest that cyclophilin D is involved in MPT induction by Ca(2+) in intact cells.
authors.affiliation: Division of Biotechnology and Life Science, Institute of Symbiotic Science and Technology, Tokyo University of Agriculture and Technology, Nakacho 2-24-16, Koganei, Tokyo, Japan.
authors.forename: Hidejiro
authors.initials: H
authors.lastname: Tazawa
authors.forename: Chisako
authors.initials: C
authors.lastname: Fujita
authors.forename: Kiyotaka
authors.initials: K
authors.lastname: Machida
authors.forename: Hiroyuki
authors.initials: H
authors.lastname: Osada
authors.forename: Yoshihiro
authors.initials: Y
authors.lastname: Ohta
mesh_terms: D000818:Animals; D002118:Calcium / Q000378:metabolism; D045744:Cell Line, Tumor; D000081406:Peptidyl-Prolyl Isomerase F; D021983:Cyclophilins / Q000235:genetics / Q000502:physiology*; D005452:Fluoresceins / Q000378:metabolism; D005456:Fluorescent Dyes / Q000378:metabolism; D015759:Ionomycin / Q000494:pharmacology; D008928:Mitochondria / Q000187:drug effects / Q000378:metabolism*; D051336:Mitochondrial Membranes / Q000187:drug effects / Q000378:metabolism*; D009154:Mutation; D019696:Peptidylprolyl Isomerase / Q000096:biosynthesis / Q000235:genetics; D010539:Permeability; D051381:Rats
chemical_list: D000081406:Peptidyl-Prolyl Isomerase F; D005452:Fluoresceins; D005456:Fluorescent Dyes; D015759:Ionomycin; D021983:Cyclophilins; D019696:Peptidylprolyl Isomerase; D002118:Calcium; C007740:fluorexon