Improvement in the in vitro maturation rate of porcine oocytes vitrified at the germinal vesicle stage by treatment with a mitochondrial permeability transition inhibitor.
In this study, we examined the effects of inhibitors of mitochondrial permeability transition (MPT), caspase activity, intracellular Ca(2+) chelator and mitochondrial Ca(2+) uniporter on survival assessed by morphological observation and in vitro maturation (IVM) of porcine vitrified germinal vesicle (GV) oocytes. When vitrified GV oocytes were matured only present in the IVM medium with an MPT inhibitor, cyclosporin A (CsA), the survival and IVM rates (36.1% and 26.8%, respectively) were significantly higher (P<0.05) than those in the other vitrified groups (10.3-12.3% and 6.2-10.3%, respectively). However, Z-VAD-fmk (Z-VAD), a caspase inhibitor, did not improve the survival and IVM rates (11.7-21.6% and 8.5-155%, respectively). When BAPTA-AM, an intracellular Ca(2+) chelator, was present in the IVM medium, the survival and IVM rates of vitrified GV oocytes (34.5-36.2% and 25.0-26.9%, respectively) were significantly higher (P<0.05) than those in the absent vitrified groups (17.2-24.2% and 12.9-19.3%, respectively). When ruthenium red (RR), an inhibitor of mitochondrial Ca(2+) uniporter, was present only in the IVM medium, the survival and IVM rates (54.5% and 39.4%, respectively) were significantly higher than those in the other vitrified groups (25.8-38.4% and 14.4-24.2%, respectively). Furthermore, blastocysts were successfully produced using porcine vitrified GV oocytes matured in the IVM medium with RR after in vitro fertilization. These results suggested that CsA, BAPTA-AM and RR but not Z-VAD have improved the survival and IVM rates of porcine vitrified GV oocytes.
Nakagawa
Shoma
S
Laboratory of Animal Breeding and Reproduction, Graduate School of Agriculture, Hokkaido University, Sapporo 060-8589, Japan.
Yoneda
Akihiro
A
Hayakawa
Koji
K
Watanabe
Tomomasa
T
D000590:Amino Acid Chloromethyl Ketones / Q000494:pharmacology; D000818:Animals; D002470:Cell Survival; D002478:Cells, Cultured; D002614:Chelating Agents / Q000494:pharmacology; D015925:Cryopreservation / Q000379:methods*; D003451:Cryoprotective Agents / Q000494:pharmacology*; D016572:Cyclosporine / Q000494:pharmacology; D015853:Cysteine Proteinase Inhibitors / Q000494:pharmacology; D004533:Egtazic Acid / Q000031:analogs & derivatives / Q000494:pharmacology; D004791:Enzyme Inhibitors / Q000494:pharmacology; D005260:Female; D008677:Metaphase; D033681:Mitochondrial Membrane Transport Proteins / Q000037:antagonists & inhibitors; D000083162:Mitochondrial Permeability Transition Pore; D009865:Oocytes / Q000166:cytology* / Q000187:drug effects*; D012430:Ruthenium Red / Q000494:pharmacology; D013552:Swine
D000590:Amino Acid Chloromethyl Ketones; D002614:Chelating Agents; D003451:Cryoprotective Agents; D015853:Cysteine Proteinase Inhibitors; D004791:Enzyme Inhibitors; D033681:Mitochondrial Membrane Transport Proteins; D000083162:Mitochondrial Permeability Transition Pore; C096713:benzyloxycarbonylvalyl-alanyl-aspartyl fluoromethyl ketone; D012430:Ruthenium Red; C070379:1,2-bis(2-aminophenoxy)ethane N,N,N',N'-tetraacetic acid acetoxymethyl ester; D004533:Egtazic Acid; D016572:Cyclosporine