OOCYTE ACTIVATION IN RHESUS MONKEY FOLLOWING INTRA CYTOPLASMIC SPERM INJECTION

胞浆内精子注射后恒河猴卵母细胞的激活

• 批准号: 6247228
• 负责人: LI MENG
• 金额: $ 4.86万
• 依托单位: OREGON REGIONAL PRIMATE RESEARCH CENTER
• 依托单位国家: 美国
• 财政年份: 1997
• 资助国家: 美国
• 起止时间: 1997-05-01 至 1998-04-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6247228
• 关键词: Primates; animal tissue; growth factor; microscopy; reproductive system

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.

胞浆内单精子注射（ICSI）提高了 辅助生殖技术的传播潜力 哺乳动物物种，并为研究提供了机会 细胞周期控制和精子诱导卵母细胞的机制 激活。 我们研究了 ICSI 对恒河猴的功效 猴子，精子注射后的受精机制和 卵母细胞激活时发生的细胞骨架重排。 这些研究是针对成熟的以及较小程度的不成熟的 卵母细胞。 射精前的精子经过清洗、获能和激活 使用传统的 ICSI 将固定化注射到卵母细胞中 方法论。 将精子注射到成熟卵母细胞中诱导卵母细胞 激活（19/86%）和原核形成。 相比之下，假 注射的卵母细胞不易激活（2/16；13%）。 本地化 卵母细胞激活因子，精子被机械分离成 头部和尾部并分别注射到成熟卵母细胞中。 87% (20/23) 的接受头部的卵母细胞发生激活。 后 尾部注射，单个微管紫苑有核，一个 在 21 个卵母细胞中的 4 个中发现原核 (PN)。 胞浆内 注射精子提取物（SE）导致卵母细胞活化 显着高于假注射的比率（76% vs 13%）。 精子诱导的卵母细胞活化也在未成熟的卵母细胞中进行了评估 中期（MI）卵母细胞； 46% (12/26) 的病例发生激活； 然而，只有 8% 的活化卵母细胞表现出 2 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 被确立为与卵母细胞相关的细胞质变化的标志物 激活。 这些结果与以下假设一致： 精子含有卵母细胞激活因子，该因子是 主要位于精子头部。 此外，还有一个激活 反应仅限于成熟卵母细胞，并伴有 细胞骨架的变化类似于传统的细胞骨架变化 施肥。