OOCYTE ACTIVATION IN RHESUS MONKEY FOLLOWING INTRA CYTOPLASMIC SPERM INJECTION

卵胞浆内单精子注射（ICSI）增加了 辅助生殖技术的潜力， 哺乳动物物种，并提供了一个机会，研究 精子诱导卵母细胞的细胞周期调控及其机制 activation. 我们研究了恒河猴ICSI的疗效 猴子，精子注射后受精的机制， 卵母细胞激活时发生的细胞骨架重排。 这些研究是在成熟的和在较小程度上不成熟的 卵母细胞 射出的精子，清洗，获能和激活前 使用常规ICSI将固定物注入卵母细胞 方法论 精子注射入成熟卵母细胞诱导卵母细胞 激活（19/86%）和原核形成。 相比之下，sham 注射的卵母细胞不容易激活（2/16;13%）。 本地化 卵母细胞活化因子，精子被机械分离成 头部和尾部，并单独注射到成熟卵母细胞中。 87%（20/23）的卵母细胞接受头部发生激活。 后 尾部注射，单个微管星形细胞成核， 21个卵母细胞中4个有原核。 浆内 注射精子提取物（SE）导致卵母细胞活化， 显著高于假注射组（76%， 13%）。 精子诱导的卵母细胞激活也在未成熟的 中期（MI）卵母细胞：46%（12/26）的病例发生激活; 但只有8%的激活卵母细胞出现2个PN。 最后， 微管蛋白在未处理和紫杉醇处理的卵母细胞中的定位是 作为与卵母细胞相关的细胞质变化的标志物 activation. 这些结果与假设一致， 精子含有卵母细胞活化因子，该因子 主要位于精子头部 此外，激活 反应仅限于成熟的卵母细胞，并伴有 细胞骨架的变化类似于常规的 受精

Intracytoplasmic sperm injection (ICSI) has increased the potential of the assisted reproductive technologies to propagate mammalian species and has provided an opportunity for research into cell cycle control and the mechanisms involved in sperm-induced oocyte activation. We have investigated the efficacy of ICSI in the rhesus monkey, the mechanism of fertilization following sperm injection and the cytoskeletal rearrangement that occurs upon oocyte activation. These studies were conducted on mature and to a lesser extent immature oocytes. Ejaculated sperm, washed, capacitated and activated before immobilization were injected into oocytes using conventional ICSI methodology. Sperm injection into mature oocytes induced oocyte activation (19/86%) and pronuclear formation. In contrast, sham injected oocytes did not activate readily (2/16;13%). To localize oocyte activation factor(s), sperm were separated mechanically into heads and tails and injected individually into mature oocytes. Activation occurred in 87% (20/23) of oocytes receiving heads. After tail injection, a single microtubule aster was nucleated and one pronucleus (PN) was seen in 4 of 21 oocytes. Intracytoplasmic injection of sperm extract (SE) resulted in oocyte activation at a significantly higher rate than that of sham injection (76% versus 13%). Sperm-induced oocyte activation was also evaluated in immature metaphase (MI) oocytes; activation occurred in 46% (12/26) of cases; however, only 8% of the activated oocytes exhibited 2 PN. Finally, -tubulin localization in untreated and taxol-treated oocytes was established as a marker for cytoplasmic changes associated with oocyte activation. These results are consistent with the hypothesis that sperm contain an oocyte activating factor(s) and the factor(s) is primarily localized in the sperm head. Moreover, an activation response is limited to mature oocytes and is accompanied by cytoskeletal changes analogous to those seen following conventional fertilization.