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Embryonic and foetal development of bovine oocytes treated with a combination of butyrolactone I and roscovitine in an enriched medium prior to IVM and IVF
Author(s) -
Ponderato Nunzia,
Crotti Gabriella,
Turini Paola,
Duchi Roberto,
Galli Cesare,
Lazzari Giovanna
Publication year - 2002
Publication title -
molecular reproduction and development
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.745
H-Index - 105
eISSN - 1098-2795
pISSN - 1040-452X
DOI - 10.1002/mrd.10134
Subject(s) - biology , germinal vesicle , andrology , embryo , blastocyst , embryogenesis , oocyte , human fertilization , maturation promoting factor , meiosis , fertilisation , insemination , anatomy , reproductive technology , pregnancy , apoptosis , biochemistry , genetics , cell cycle , cyclin dependent kinase 1 , medicine , gene
Abstract Cattle oocytes were maintained at germinal vesicles (GV) stage for 24 hr using a combination of two specific and potent inhibitors of M‐phase promoting factor (MPF) kinase activity, butyrolactone I (BL‐I) and roscovitine (ROS). The media used for inhibition were (a) TCM‐199 only and (b) TCM‐199 supplemented with serum, hormones and growth factors. The effective doses of inhibitors were 6.25 μM BL‐I and 12.5 μM ROS in medium (a) and 50 μM BL‐I and 12.5 μM ROS in medium (b). After inhibition, about 90% of the oocytes resumed meiosis and reached the metaphase II (MII) stage during 24 hr of maturation. Following fertilisation the percentage of cleavage (D +2), compacted morula (D +6), blastocysts on D +7 and D +8 and the survival to freezing and thawing of grade 1 embryos frozen on D +7 were not different between the experimental treated groups and the control. In order to evaluate early foetal development, two groups of five grade 1 D +7 blastocysts derived from treated oocytes and two groups of five control embryos were transferred nonsurgically in four synchronised recipient heifers. On D +27, the recipients were slaughtered and the foetuses were recovered. In both groups, six foetuses developed out of the 10 embryos transferred. In conclusion, several supplements can be added to the prematuration medium of bovine oocytes without reducing the quality of inhibition but also without improving their subsequent developmental competence versus treated oocytes in TCM‐199 only and versus untreated control. Furthermore, the prematuration step used in this study does not interfere with normal foetal development during the first stages of organogenesis. Mol. Reprod. Dev. 62:513–518, 2002. © 2002 Wiley‐Liss, Inc.