Department of Veterinary Physiology and Pharmacology, College of Veterinary Medicine and Biomedical Sciences, Texas A&M University, College Station, TX 77843-4466, USA.
Mol Reprod Dev. 2010 Aug;77(8):651-61. doi: 10.1002/mrd.21186.
There is currently much interest in assisted reproduction techniques in the horse, however, many aspects of oocyte maturation, fertilization, and embryo development in the horse differ from those in other species. Because of the close attachment of the equine oocyte to the follicle wall, scraping of the follicle is the most effective method for oocyte recovery. A notable feature of equine oocytes is that those with expanded cumuli (Ex oocytes), which originate from atretic follicles, have higher meiotic competence (ability to mature to metaphase II in vitro) than do oocytes with compact cumuli (Cp oocytes). Cp oocytes originate in viable follicles but are largely juvenile. Recovery and culture of equine oocytes immediately after slaughter yields a higher maturation rate than that obtained from oocytes after ovary storage; this is related to damage to chromatin in Cp oocytes during storage. In contrast, developmental competence (rate of blastocyst development in vitro) is higher in oocytes recovered from the ovary after a delay. The optimum duration of maturation varies based on cumulus morphology and time of recovery from the ovary, but there is no difference in developmental competence between Ex and Cp oocytes. Because standard in vitro fertilization is not repeatable in the horse, oocyte transfer (surgical transfer of oocytes to the oviducts of inseminated mares) has been developed to allow fertilization of isolated oocytes. Fertilization in vitro may be achieved using intracytoplasmic sperm injection; culture of injected oocytes in a medium with high glucose can yield over 30% blastocyst development.
目前,人们对马的辅助生殖技术非常感兴趣,然而,马的卵母细胞成熟、受精和胚胎发育的许多方面与其他物种不同。由于马的卵母细胞与卵泡壁紧密相连,刮取卵泡是回收卵母细胞最有效的方法。马卵母细胞的一个显著特征是,那些扩张的卵丘(Ex 卵母细胞),来源于闭锁卵泡,具有更高的减数分裂能力(在体外成熟至中期 II 的能力),而那些紧密的卵丘(Cp 卵母细胞)则没有。Cp 卵母细胞起源于有活力的卵泡,但大部分是幼稚的。在屠宰后立即回收和培养马的卵母细胞比从卵巢储存的卵母细胞获得的成熟率更高;这与 Cp 卵母细胞在储存过程中染色质的损伤有关。相比之下,延迟从卵巢中回收的卵母细胞的发育能力(体外囊胚发育率)更高。最佳成熟时间因卵丘形态和从卵巢中回收的时间而异,但 Ex 和 Cp 卵母细胞之间的发育能力没有差异。由于马的标准体外受精不可重复,因此开发了卵母细胞转移(将卵母细胞手术转移到已受精母马的输卵管中),以允许对分离的卵母细胞进行受精。体外受精可以通过胞质内精子注射来实现;在高葡萄糖的培养基中培养注射的卵母细胞可以产生超过 30%的囊胚发育。