Milrinone Treatment of Bovine Oocytes During In Vitro Maturation Benefits Production of Nuclear Transfer Embryos by Improving Enucleation Rate and Developmental Competence

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概要

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• In the production of cattle nuclear transfer embryos, the production efficiency is affected by the oocyte developmental competence and successful enucleation rate. This study investigated the effect of treating oocytes with milrinone, a phosphodiesterase inhibitor, on these two characteristics. When cumulus–oocyte complexes (COCs) were cultured for 19 h with 0, 50 or 100 μM of milrinone, the enucleation rate was significantly improved by 100 μM milrinone. However, milrinone treatment during in vitro maturation (IVM) also delayed meiotic progression by at least 2 h, which would affect the examination of enucleation rate and developmental competence of oocytes. Thus, in the second experiment, meiotic resumption was temporarily inhibited with butyrolactone I (BL-I; 100 μM, 18 h) to decrease the delayed maturation caused by milrinone; this enabled a more accurate comparison of the effects of milrinone after oocyte maturation. In nuclear transfer embryo production, oocytes treated with milrinone (100 μM, 20 h) showed a significantly higher rate of enucleation compared with that of control oocytes. This improved enucleation rate was associated with a closer location of the metaphase plate to the first polar body in the treated oocytes compared with that in control oocytes. Furthermore, milrinone improved the frequency of development to the blastocyst stage in the resulting embryos. In conclusion, milrinone supplementation during IVM improved enucleation rates by rendering the metaphase plate in close proximity to the first polar body, and this treatment also improved oocyte developmental competence. These benefits additively improved the yield of cloned embryos that developed to the blastocyst stage.

• 日本繁殖生物学会の論文

著者

• Iga Kosuke

  Reproductive Biology And Technology Research Subteam National Agricultural Research Center For Tohok

• NARUSE Kenji

  NARO Tohoku Agricultural Research Center, Iwate 020-0198, Japan

• IGA Kosuke

  NARO Tohoku Agricultural Research Center, Iwate 020-0198, Japan

• SHIMIZU Manabu

  NARO Tohoku Agricultural Research Center, Iwate 020-0198, Japan

• TAKENOUCHI Naoki

  NARO Tohoku Agricultural Research Center, Iwate 020-0198, Japan

• AKAGI Satoshi

  NARO Institute of Livestock and Grassland Science, Ibaraki 305-0901, Japan

• SOMFAI Tamas

  NARO Institute of Livestock and Grassland Science, Ibaraki 305-0901, Japan

• HIRAO Yuji

  NARO Tohoku Agricultural Research Center, Iwate 020-0198, Japan

関連論文

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• Possible Involvement of Insulin-like Growth Factor-I in Mediating the Stimulatory Effect of Recombinant Bovine Growth Hormone on Maturation of Bovine Oocytes In Vitro
• Recombinant Bovine Growth Hormone Stimulates Nuclear Maturation of Bovine Oocytes in Vitro and Promotes Subsequent Embryonic Development
• Optimization of Oxygen Concentration for Growing Bovine Oocytes In Vitro : Constant Low and High Oxygen Concentrations Compromise the Yield of Fully Grown Oocytes
• Milrinone Treatment of Bovine Oocytes During In Vitro Maturation Benefits Production of Nuclear Transfer Embryos by Improving Enucleation Rate and Developmental Competence
• Optimization of Oxygen Concentration for Growing Bovine Oocytes In Vitro: Constant Low and High Oxygen Concentrations Compromise the Yield of Fully Grown Oocytes
• Milrinone Treatment of Bovine Oocytes During In Vitro Maturation Benefits Production of Nuclear Transfer Embryos by Improving Enucleation Rate and Developmental Competence

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