Contraceptive agents to block sperm function in fertiliz

描述(申请者提供)：本项目重点研究避孕药具的开发 受精剂阻断两步受精。这些药物将是口服的药物，可以阻止精子与卵子的透明带结合，或者阻止精子进入输卵管。 为了开发第一种避孕药，我们将鉴定精子表面蛋白，使精子能够与卵子的透明带结合。尽管在过去的20年里进行了深入的研究，但具有这一功能的精子蛋白仍不清楚。在我们最近的研究中，我们开发了两个新的突变小鼠品系，在这些品系中，雄性不育，并产生无法与小带结合的精子。这些基因敲除小鼠的可获得性，以及新的基因组和蛋白质组技术，为我们提供了一个全新的方法来解决这个问题。在鉴定了精子的小带结合蛋白后，我们将建立一种高通量测试来筛选阻止人类小带结合蛋白与人ZP3结合的抑制剂。这种高通量筛查将是允许与工业合作伙伴进行后续避孕开发的关键。 对于第二种避孕药，起点将是我们的一种携带精子黏附蛋白受精素a缺失的小鼠品系。缺乏受精素a的雄性小鼠产生的精子无法进入输卵管。我们将测试这样一种假设，即精子与子宫或输卵管交界处上皮细胞的粘连需要受精素a，而这一结合步骤会产生一个信号，使精子有能力进入输卵管。我们建议检测受精素与上皮细胞的粘附性，并筛选会导致子宫内精子停滞的小分子粘附剂。

DESCRIPTION (provided by applicant): This project focuses on the development of contraceptive agents to block two steps in fertilization. These agents will be orally available drugs that block either sperm binding to the egg's zona pellucida or sperm entry into the oviduct. To develop the first contraceptive drug, we will identify the sperm surface protein that enables sperm to bind to the egg's zona pellucida. Despite intensive investigation for the last 20 years, the sperm protein with this function remains unknown. In our recent research, we have developed two new mutant mouse lines in which the males are infertile and make sperm that fail to bind to the zona. The availability of these knockout mice, along with new genomic and proteomic technologies, provides us a completely novel approach to this problem. Having identified the sperm's zona binding protein, we will establish a high-throughput assay to screen inhibitors that block binding of the human zona binding protein to human ZP3. This high-throughput screen will be the key that allows subsequent contraceptive development with an industrial partner. For the second contraceptive agent, the starting point will be one of our mouse lines that carries a deletion of the sperm adhesion protein fertilin a. Male mice lacking fertilin a produce sperm that fail to enter the oviduct. We will test the hypothesis that sperm adhesion to the uterine or uterotubal junction epithelium requires fertilin a and that this binding step results in a signal that makes sperm competent to enter the oviduct. We propose assays for fertilin a adhesion to the epithelium and for screening small molecule inhibitors of the adhesion that will cause sperm arrest in the uterus.