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Effect of Temporary Meiotic Attenuation of Oocytes with Butyrolactone I and Roscovitine in Resistance to Bovine Embryos on Vitrification
Author(s) -
Maziero RRD,
Guaitolini CRF,
Paschoal DM,
Kievitsbosch T,
Guastali MD,
Moraes CN,
LandimAlvarenga FC
Publication year - 2016
Publication title -
reproduction in domestic animals
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.546
H-Index - 66
eISSN - 1439-0531
pISSN - 0936-6768
DOI - 10.1111/rda.12667
Subject(s) - vitrification , andrology , meiosis , embryo , biology , microbiology and biotechnology , medicine , genetics , gene
Contents This study aimed to produce in vitro bovine embryos by the addition of two drugs, which is responsible for oocyte meiosis inhibition: roscovitine ( ROS ) and butyrolactone I ( BL ‐I). Oocytes were recovered from slaughtered cows and matured in a commercial medium and maintained in a 5% CO 2 atmosphere. Oocytes were maintained for 6 h in an in vitro maturation ( IVM ) medium containing ROS (12.5 μ m ), BL ‐I (50 μ m ) and association of drugs ( ROS 6.25 μ m and BL ‐I 25 μ m ). Oocytes were cultured for 18 h in an agent‐free medium for the resumption of meiosis. After 24 h of maturation, oocytes were inseminated in the commercial in vitro fertilization ( IVF ) medium. Presumptive zygotes were cultured in SOF aa medium in a 5% CO 2 atmosphere. On day 3, rate of cleavage was evaluated and on days 6 and 7, rate of blastocyst formation. BL ‐I and its association with the ROS increased the rates of cleavage and blastocyst formation (p < 0.05). The ROS alone was inefficient, impairing embryonic development, with low rates of blastocyst formation when compared to the control group and other treatments (p < 0.05). The embryos from BL ‐I and ROS + BL ‐I groups presented higher number of cells and lower rates of cellular apoptosis compared to other groups, either for the fresh or for post‐thawing embryos. Embryos from ROS + BL ‐I group showed to be more resistant to the vitrification process, presenting a higher rate of embryonic re‐expansion (p < 0.05). In conclusion, block of meiosis using BL ‐I or its association with ROS increased the rate of blastocyst formation, and the association of ROS + BL ‐I resulted in a better resistance to the embryo cryopreservation process.