精子gACE在精卵结合过程中作用的分子机制研究及临床意义

精卵结合对于体内受精以及常规IVF中的体外受精都是至关重要的一环。然而目前其分子机制仍不明确,这也是导致常规IVF中仍时有受精失败发生的主要原因。我们前期研究发现男性患者精子gACE表达缺乏导致常规IVF中完全受精失败和低受精率的发生。因此我们假设:gACE可能在精卵结合过程中发挥重要功能,并可能作为评估男性生育能力的重要分子指标。本研究拟在前期研究的基础上通过多种分子生物学方法进一步研究精浆和精子中gACE表达水平,以期评估gACE能否作为男性生育能力重要分子指标。同时本课题将研究gACE候选下游蛋白的作用,并应用基因和蛋白芯片技术寻找和确定上下游调控分子和作用靶点,并用质粒共转染及体外透明带诱导顶体反应的方法验证;本课题还将筛选gACE特异性调控区域,并通过双荧光素酶报告检测系统以及体外转录系统分析其对转录活性的影响。研究结果将深化对精卵结合的认识,并对预防受精失败的发生具有指导意义。

Sperm-egg binding is key to in vivo fertilization and in vitro fertilization (IVF). However, its molecular mechanisms remain largely unknown, which explains partly why fertilization failure can still occur in conventional IVF cycles. Our previous study suggested that human sperm devoid of gACE induced total fertilization failure and low fertilization rate by conventional IVF. Therefore, we can assume that gACE might play a vital role in sperm-egg binding process and might be used as a critical molecular marker to evaluate male fertility. The expression level of gACE in human sperm and seminal plasma will be investigated by different molecular biological techniques. Gene and protein chip technology will be applied to screen upstream and downstream regulatory molecules and potential targets of gACE, further confirmed by co-transfection of plasmid and ZP induced acrosome reaction in vitro. Finally, dual luciferase detection and in vitro transcription system will be applied to analyze the influence on the transcriptional activity of specific regulatory regions of gACE. This study would deepen the understanding of sperm-egg binding process, meanwhile, it can also help to prevent fertilization failure and diagnose causes of male infertility.