C-Ros pathways as targets for contraceptive development

描述（由申请人提供）：附睾在精子成熟中起着至关重要的作用，因此被认为是开发男性避孕药的主要目标。我们的策略是针对受体，激酶和磷酸酶在附睾是至关重要的男性生育能力。这些靶标是理想的，因为它们适用于小分子量抑制剂，并且血液-附睾屏障不会成为此类抑制剂的强大障碍。考虑到这些策略，我们的目标是研究孤儿酪氨酸激酶受体c-Ros作为男性避孕药开发的目标。我们也对确定c-Ros的上游调控成分和下游目标作为男性避孕药开发的目标感兴趣。选择C-Ros是因为敲除C-Ros的雄性小鼠是不育的。本课题的具体目的是：(1)验证通过调节c-Ros的表达和活性调节成年雄性小鼠生育能力的假说。为了实现这一特定目标，我们将在小鼠的初始片段中产生一种新的条件c-Ros敲除。(2)为了验证c-Ros的活性取决于其激酶结构域与其他结合蛋白（包括新型和已知的激酶和磷酸酶）的结合的假设。与c-Ros活性相关的蛋白质将被视为开发男性避孕药的目标。我们将产生初始片段特异性条件敲除SHP-1磷酸酶，该磷酸酶与c-Ros的激酶结构域结合。(3)验证c-Ros的活性依赖于其胞外结构域与膜结合受体的关联的假设。我们将产生一个初始的片段特异性条件敲除代谢性谷氨酸受体样孤儿g蛋白偶联受体，这是假定的c-Ros配体。下游激酶、磷酸酶和受体在每个假定的不育敲除动物的初始片段中的表达和活性变化将被确定并进一步考虑作为避孕开发的目标。因此，根据RFA的研究范围，我们将进行“精子成熟过程的研究，以确定具体目标，参与男性生育能力的控制。”

DESCRIPTION (provided by applicant): The epididymis plays a crucial role in the maturation of spermatozoa and is, therefore, considered to be a prime target for the development of a male contraceptive. Our strategy is to target receptors, kinases and phosphatases within the epididymis that are crucial for male fertility. These targets are ideal because they are amenable to small molecular weight inhibitors and the blood-epididymis barrier would not be a formidable hurdle for such inhibitors. With these strategies in mind, our goal is to examine the orphan tyrosine kinase receptor c-Ros as a target for male contraceptive development. We are also interested in identifying the components of upstream regulators and downstream targets of c-Ros as targets for male contraceptive development. C-Ros was chosen because c-Ros knockout male mice are infertile. The specific aims of this proposal are: (1) To test the hypothesis that modulating the expression and activity of c-Ros can regulate fertility in adult male mice. To accomplish this specific aim we will generate a novel conditional c-Ros knockout in the initial segment of mice. (2) To test the hypothesis that the activity of c-Ros is dependent upon the association of its kinase domain with other binding proteins including novel and known kinases and phosphatases. Proteins that associate with c-Ros activity will be considered as targets for the development of a male contraceptive. We will generate an initial segment-specific conditional knockout of the SHP-1 phosphatase, which binds to the kinase domain of c-Ros. (3) To test the hypothesis that the activity of c-Ros is dependent upon its association of its extracellular domain with a membrane-bound receptor. We will generate an initial segment-specific conditional knockout of the metabotropic glutamate receptor-like orphan G-protein coupled receptor, which is the putative ligand for c-Ros. Expression and activity of downstream kinases, phosphatases and receptors that change in the initial segment of each putative infertile knockout animal will be identified and considered further as targets for contraceptive development. Therefore, in accordance with the research scope of the RFA, we will perform "research on the processes of sperm maturation with the goal of defining specific targets, involved in the control of male fertility."