title: Rescue of vitrified-warmed bovine oocytes with rho-associated coiled-coil kinase inhibitor.
abstract: Cryotolerance of matured bovine oocytes is not fully practical even though a promising vitrification procedure with a ultrarapid cooling rate was applied. The present study was conducted to investigate whether recovery culture of vitrified-warmed bovine oocytes with an inhibitor (Y-27632) of Rho-associated coiled-coil kinase (ROCK) can improve the developmental potential after in vitro fertilization (IVF) and in vitro culture. Immediately after warming, almost all oocytes appeared to be morphological normal. Treatment of the postwarming oocytes with 10 μM Y-27632 for 2 h resulted in the significantly higher oocyte survival rate before IVF as well as higher cleavage rate and blastocyst formation rate. Quality analysis of the resultant blastocysts in terms of total cell number and apoptotic cell ratio also showed the positive effect of the Y-27632 treatment. Time-dependent change in mitochondrial activity of the vitrified-warmed oocytes was not influenced by ROCK inhibition during the period of recovery culture. However, the ability of ooplasm to support single-aster formation was improved by the ROCK inhibition. Thus, inhibition of ROCK activity in vitrified-warmed bovine oocytes during a short-term recovery culture can lead to higher developmental competence, probably due to decreased apoptosis and normalized function of the microtubule-organizing center.
authors.affiliation: Interdisciplinary Graduate School of Science and Technology, Shinshu University, Ueda, Nagano, Japan.
authors.forename: In-Sul
authors.initials: IS
authors.lastname: Hwang
authors.forename: Hiromasa
authors.initials: H
authors.lastname: Hara
authors.forename: Hak-Jae
authors.initials: HJ
authors.lastname: Chung
authors.forename: Masumi
authors.initials: M
authors.lastname: Hirabayashi
authors.forename: Shinichi
authors.initials: S
authors.lastname: Hochi
mesh_terms: D000577:Amides / Q000494:pharmacology*; D000818:Animals; D017209:Apoptosis / Q000187:drug effects; D001755:Blastocyst / Q000166:cytology / Q000502:physiology; D002417:Cattle; D015925:Cryopreservation / Q000379:methods*; D022101:Microtubule-Organizing Center / Q000187:drug effects / Q000502:physiology; D008928:Mitochondria / Q000187:drug effects / Q000502:physiology; D009865:Oocytes / Q000166:cytology / Q000187:drug effects* / Q000502:physiology; D011725:Pyridines / Q000494:pharmacology*; D058989:Vitrification; D054460:rho-Associated Kinases / Q000037:antagonists & inhibitors*
chemical_list: D000577:Amides; D011725:Pyridines; C108830:Y 27632; D054460:rho-Associated Kinases
keywords: Cryotop vitrification; ROCK inhibitor; apoptosis; bovine oocytes; cryopreservation; cytoskeleton; microtubule assembly; oocyte; vitrification