Agung Budiyanto, Otoi Takeshige, Wongsrikeao Pimprapar, Taniguchi Masayasu, Shimizu Ryohei, Watari Hajime, Nagai Takashi
Laboratory of Animal Reproduction, Department of Veterinary Sciences, Yamaguchi University, Yamaguchi, Japan.
J Reprod Dev. 2006 Feb;52(1):123-7. doi: 10.1262/jrd.17055. Epub 2005 Nov 7.
It has been suggested that the maturational stage of oocytes at time of insemination influences the sex ratio of resulting embryos. However, there are very few reports concerning the relationship between the maturation culture period of oocytes and the sex ratio of resulting embryos. The objective of this study was to investigate the effects of in vitro maturation culture period for bovine oocytes on the sex ratio of in vitro produced blastocysts using a novel technique of loop-mediated isothermal amplification (LAMP). Cumulus-oocyte complexes were collected from the ovaries of slaughtered cows, and then matured in vitro for various periods (16, 22, 28, and 34 h). After maturation culture for each period, the oocytes were inseminated with frozen-thawed spermatozoa, and then cultured in vitro. Blastocysts were harvested on Day 7 after insemination, and the sex of the embryos was examined using the LAMP method. The rates of oocytes matured to the metaphase II stage were significantly lower (P < 0.05) in the 16-h maturation group than in the other groups. The proportion of blastocyst formation after insemination was significantly higher (P < 0.05) in the 22-h maturation group than in the other groups. The proportion of male blastocysts increased with the increase in maturation culture period. The proportion of male blastocysts derived from oocytes matured for 34 h was significantly higher (P < 0.05) than from oocytes matured for 16 and 22 h. These results indicate that the sex ratio of in vitro fertilized embryos is apparently influenced by the maturation culture period of the oocytes.
有人提出,授精时卵母细胞的成熟阶段会影响所产生胚胎的性别比例。然而,关于卵母细胞成熟培养时间与所产生胚胎性别比例之间关系的报道非常少。本研究的目的是使用一种新型的环介导等温扩增(LAMP)技术,研究牛卵母细胞的体外成熟培养时间对体外产生的囊胚性别比例的影响。从屠宰母牛的卵巢中收集卵丘-卵母细胞复合体,然后在体外培养不同时间(16、22、28和34小时)。在每个时间段成熟培养后,用冷冻解冻的精子对卵母细胞进行授精,然后进行体外培养。在授精后第7天收获囊胚,并用LAMP方法检查胚胎的性别。16小时成熟组中成熟到中期II期的卵母细胞比例显著低于其他组(P<0.05)。22小时成熟组授精后的囊胚形成比例显著高于其他组(P<0.05)。雄性囊胚的比例随着成熟培养时间的增加而增加。成熟34小时的卵母细胞来源的雄性囊胚比例显著高于成熟16和22小时的卵母细胞来源的雄性囊胚比例(P<0.05)。这些结果表明,体外受精胚胎的性别比例明显受卵母细胞成熟培养时间的影响。