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Reproduction, Fertility and Development Reproduction, Fertility and Development Society
Vertebrate reproductive science and technology
RESEARCH ARTICLE

Synchronisation of canine germinal vesicle stage oocytes prior to in vitro maturation alters the kinetics of nuclear progression during subsequent resumption of meiosis

Carol Hanna A , Suzanne Menges A , Duane Kraemer A and Charles R. Long A B
+ Author Affiliations
- Author Affiliations

A Texas A and M University, College of Veterinary Medicine, Department of Physiology and Pharmacology, College Station, TX 77843-4466, USA.

B Corresponding author. Email: clong@cvm.tamu.edu

Reproduction, Fertility and Development 20(5) 606-614 https://doi.org/10.1071/RD07227
Submitted: 20 December 2007  Accepted: 7 April 2008   Published: 21 May 2008

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 μmol 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 μmol 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.


Acknowledgements

The authors of this paper would like thank Kim Green and JoAnne Stokes for technical assistance and acknowledge Bio-Arts Research Corporation, Mill Valley, California, USA, for providing funding for this project.


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