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Effects of Roscovitine on Nuclear Maturation, Spindle Configuration, and Chromosome Alignment in Porcine Oocytes  

Park, Sang-Hyoun (Laboratory of Veterinary Obstetrics & Reproductive Biotechnologies, College of Veterinary Medicine & Bio-safety Research Institute, Chonbuk National University)
Yu, Il-Jeoung (Laboratory of Veterinary Obstetrics & Reproductive Biotechnologies, College of Veterinary Medicine & Bio-safety Research Institute, Chonbuk National University)
Publication Information
Journal of Embryo Transfer / v.25, no.4, 2010 , pp. 251-258 More about this Journal
Abstract
In the present study, effects of concentration and time of culture in presence of roscovitine on nuclear maturation and meiotic spindle configuration, chromosomal alignment were examined in porcine oocytes. In experiment 1, porcine cumulus oocyte complexes (COCs) were cultured at $39^{\circ}C$ in a 5% $CO_2$ atmosphere in North Carolina State University 23 (NCSU-23) supplemented with 25, 50, 75 or $100\;{\mu}M$ roscovitine for 22 h and then were cultured for additional 22 h after removal of roscovitine. Nuclear maturation and morphology of the meiotic spindle and chromosomal alignment were examined to determine the optimal concentration of roscovitine in oocyte maturation. In experiment 2, COCs were cultured in NCSU-23 supplemented with $50\;{\mu}M$ roscovitine for 17, 20, 27 or 42 h and then an additional 22 h without roscovitine was followed to determine the optimal time of culture. The optimal concentration of roscovitine to arrest and resume meiosis of porcine oocyte was $50\;{\mu}M$ by examining nuclear status (p<0.05) and normal spindle and chromosome configuration. The optimal time of culture in presence of roscovitine to arrest meiosis of porcine oocyte was 17 h (p<0.05), although MII rates and normal morphology of the meiotic spindle and chromosomal alignment were not significantly different among various times of culture. In conclusion, the optimal concentration and time of culture in presence of roscovitine to arrest porcine oocytes are $50\;{\mu}M$ and 17 h, respectively.
Keywords
roscovitine; nuclear maturation; meiotic spindle configuration; chromosome alignment;
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