MOLECULAR MECHANISM AND DEVELOPMENT CONSEQUENCES OF ICSI

辅助生殖技术（ART）的实践，以克服 在过去的两年中，人类的不孕症急剧增加， 几十年来，并最终广泛使用胞浆内精子 注射（ICSI），可以克服以前的不孕症的情况 被认为是棘手的。由于执行的固有困难， 基础研究与有限的人力物力和政府 禁止对人类卵子和植入前胚胎进行基础研究， 抗逆转录病毒疗法的发展基本上是在缺乏基本的 使用动物模型系统进行研究。从本质上讲，这是一个 艺术先于科学在这份资助申请中，我们建议使用鼠标 作为研究分子机制的模型系统， ICSI的发育/行为后果。鸡蛋的能力 激活的孤雌生殖刺激增加与时间后hCG 给药并与中期II-阻滞的进展相关 将卵转化为类似于间期的状态。第一个具体目标将测试 假设，类似于孤雌卵激活， hCG后ICSI诱导的卵子激活随时间增加 局虽然胞浆内精子注射已经彻底改变了 男性不育，ICSI如何激活卵子的分子基础， 启动早期发育的程序是知之甚少的， 特别是考虑到ICSI绕过了正常的使用途径， 使卵子受精。了解的分子基础 通过ICSI激活卵子将不可避免地了解 针对其失效原因和合理性，提出了提高其有效性的途径。的 第二个具体目标将测试ICSI招募信号传导的假设 在自然过程中通常被激活的途径 受精，并导致卵子的正常补充 activation.胞浆内单精子注射作为一种选择方法的迅速普及， 克服人类男性不育引起了激烈的争论， 对后代造成的长期风险。虽然 初看之下，没有明显的显著增加， ICSI衍生的先天性大体异常的发生率 后代，这些孩子还年轻，因此更微妙的影响， 影响行为和智力发展的因素 很明显第三个具体目标将测试小鼠ICSI的假设， 会导致孩子的发育和行为异常， 产生的后代。

The practice of Assisted Reproductive Technology (ART) to overcome infertility in humans has increased dramatically during the past two decades and has culminated in the widespread use of intracytoplasmic sperm injection (ICSI), which can overcome instances of infertility previously thought to be intractable. Due to the inherent difficulty of conducting basic research with the limited amounts of human material and governmental bans on basic research of human eggs and pre-implantation embryos, the development of ART hs occurred essentially in the absence of basic research using an animal model system. In essence, this is an example of ART before science. In this grant application, we propose to use the mouse as a model system to study the molecular mechanism and developmental/behavioral consequences of ICSI. The ability of eggs to be activated by parthenogenetic stimuli increases with the time post-hCG administration and correlates with progression of metaphase II-arrested eggs into an interphase-like state. The first Specific Aim will test the hypothesis that, similar to parthenogenetic egg activation, the success of ICSI-induced egg activation increases with time following hCG administration. While ICSI has clearly revolutionized the treatment of male infertility, the molecular basis of how ICSI activates the egg and initiates the program of early development is poorly understood, especially in light of the fact that ICSI bypasses the normal pathway used by the sperm to fertilize the egg. Understanding the molecular basis of egg activation by ICSI will inevitably yield an understanding as to causes for its failure and rationale approaches to enhances its efficacy. The second Specific Aim will test the hypothesis that ICSI recruits signaling pathways that are normally activated during the course of natural fertilization and results in the normal complement of events of egg activation. The rapidly gaining use of ICSI as the method-of-choice to overcome human male infertility has raised a vigorous debate concerning the long-term risk imposed to the resultant offspring. Although it is reassuring at first glance that there is no apparent significant increase in the incidence of gross congenital abnormalities in ICSI-derived offspring, these children are still young and hence more subtle effects that influence behavioral and intellectual development may not yet be apparent. The third Specific Aim will test the hypothesis that mouse ICSI results in subtle developmental and behavior abnormalities in the generated offspring.