Lee Seung Tae, Han Ho Jae, Oh Seo Jin, Lee Eun Ju, Han Jae Yong, Lim Jeong M
Department of Food and Animal Biotechnology, Seoul National University, Seoul, Korea.
Mol Reprod Dev. 2006 Aug;73(8):1022-33. doi: 10.1002/mrd.20500.
This study was undertaken to determine the effects of gonadotrophin on cytoskeletal dynamics and embryo development and its role in improving the retrieval of developmentally competent oocytes. Female golden hamsters were injected with human chorionic gonadotrophin (hCG; 5-, 7.5- or 15-IU) on the day 4 of estrus, pregnant mare serum gonadotrophin (PMSG; 5-, 7.5- or 15-IU) on the day 1 of estrus, or 15-IU hCG at 56 hr post-15-IU PMSG injection in any cycle except estrus. Increasing the hCG dose decreased not only retrieval rate of 2-cell embryo but development to blastocyst after subsequent in vitro culture. Whereas, although increasing the PMSG dose induced increasing the number of 2-cell embryo and blastocyst, 15-IU PMSG injection caused retardation of development to blastocyst. No 2-cell embryos were retrieved by injecting both PMSG and hCG. The injections of 15-IU hCG and 7.5- or 15-IU PMSG inhibited the proliferation of trophectodermal and inner cell mass cells, respectively. Gonadotrophin injection didn't influence microtubular spindle formation, but 5- or 15-IU hCG, 15-IU PMSG, or PMSG and hCG injections induced aberrant cortical granule (CG) and microfilament distribution. After 15-IU hCG or PMSG and hCG injections, fewer oocytes had enriched cortical actin domains, and the expression of alpha-, beta- and gamma-actin genes was greatly increased. In conclusion, a high dose of gonadotrophins alters the microfilament and CG distribution, which in turn reduces the developmental competence of oocytes. Injecting a reduced dose of PMSG to initiate ovarian hyperstimulation without triggering ovulation contributes to the efficient retrieval of developmentally competent oocytes.
本研究旨在确定促性腺激素对细胞骨架动力学和胚胎发育的影响及其在提高发育潜能良好的卵母细胞回收率方面的作用。在发情期第4天给雌性金黄仓鼠注射人绒毛膜促性腺激素(hCG;5、7.5或15国际单位),在发情期第1天注射孕马血清促性腺激素(PMSG;5、7.5或15国际单位),或在除发情期外的任何周期中,在注射15国际单位PMSG后56小时注射15国际单位hCG。增加hCG剂量不仅会降低2细胞胚胎的回收率,还会降低随后体外培养后的囊胚发育率。然而,虽然增加PMSG剂量会导致2细胞胚胎和囊胚数量增加,但注射15国际单位PMSG会导致囊胚发育延迟。同时注射PMSG和hCG未回收2细胞胚胎。注射15国际单位hCG和7.5或15国际单位PMSG分别抑制滋养外胚层和内细胞团细胞的增殖。促性腺激素注射不影响微管纺锤体形成,但5或15国际单位hCG、15国际单位PMSG或PMSG与hCG联合注射会诱导皮质颗粒(CG)和微丝分布异常。注射15国际单位hCG或PMSG与hCG联合后,具有丰富皮质肌动蛋白结构域的卵母细胞减少,α-、β-和γ-肌动蛋白基因的表达大幅增加。总之,高剂量促性腺激素会改变微丝和CG分布,进而降低卵母细胞发育潜能。注射低剂量PMSG启动卵巢超刺激而不触发排卵有助于有效回收发育潜能良好的卵母细胞。