title: Synchronisation of canine germinal vesicle stage oocytes prior to in vitro maturation alters the kinetics of nuclear progression during subsequent resumption of meiosis.
abstract: Inhibition of meiosis before in vitro maturation (IVM) can improve meiotic competence in immature mammalian oocytes. Therefore, meiosis-inhibiting agents were evaluated singularly for the ability to arrest and synchronise germinal vesicle (GV) stage canine oocytes, and the most effective treatments were combined to improve meiotic resumption rates. Oocytes cultured in 2 ng mL(-1) oestradiol (E2), 10 IU mL(-1) eCG, or both (EG) for 72 h resulted in significantly fewer oocytes resuming meiosis in EG than the control, E2, or with eCG. Oocytes cultured in 50 or 100 micromol L(-1) of butyrolactone 1 or roscovitine (ROS) for up to 48 h did not resume meiosis nor increase subsequent meiotic resumption rates following IVM. A combination of 50 micromol L(-1) ROS and EG treatment for 48 h significantly increased the proportion of canine oocytes in meiotic arrest. More importantly, following 48 h of IVM, ROS+EG-treated oocytes demonstrated a dramatic increase in the ability to resume meiosis compared with the non-treated controls (51.3 +/- 8.2% and 10.8 +/- 4.5%, respectively; P < 0.05). These data indicate that chemical and biological meiotic inhibitors are effective at inducing GV arrest in canine oocytes. Furthermore, these inhibitors are reversible and beneficial to subsequent meiotic resumption in vitro.
authors.affiliation: Texas A and M University, College of Veterinary Medicine, Department of Physiology and Pharmacology, College Station, TX 77843-4466, USA.
authors.forename: Carol
authors.initials: C
authors.lastname: Hanna
authors.forename: Suzanne
authors.initials: S
authors.lastname: Menges
authors.forename: Duane
authors.initials: D
authors.lastname: Kraemer
authors.forename: Charles R
authors.initials: CR
authors.lastname: Long
mesh_terms: D015107:4-Butyrolactone / Q000031:analogs & derivatives / Q000494:pharmacology; D000818:Animals; D050256:Antimitotic Agents / Q000494:pharmacology*; D018929:Cell Culture Techniques; D002467:Cell Nucleus / Q000187:drug effects* / Q000235:genetics; D002478:Cells, Cultured; D004285:Dogs / Q000502:physiology*; D004338:Drug Combinations; D004958:Estradiol / Q000494:pharmacology; D005260:Female; D006064:Gonadotropins, Equine / Q000494:pharmacology; D008540:Meiosis / Q000187:drug effects*; D054315:Oocyte Retrieval / Q000009:adverse effects / Q000379:methods; D009865:Oocytes / Q000187:drug effects* / Q000254:growth & development; D009866:Oogenesis / Q000187:drug effects* / Q000502:physiology; D011687:Purines / Q000494:pharmacology; D000077546:Roscovitine
chemical_list: D050256:Antimitotic Agents; D004338:Drug Combinations; D006064:Gonadotropins, Equine; D011687:Purines; D000077546:Roscovitine; D004958:Estradiol; C082868:butyrolactone I; D015107:4-Butyrolactone