FAS ANTIGEN AND OVARIAN CELL APOPTOSIS

总体目标是确定触发事件和分子机制 调节卵巢细胞的程序性细胞死亡。 Fas抗原是一种 触发凋亡的跨膜受体（一种程序化细胞 死亡）。 该项目将明确Fas抗原在卵巢癌中的作用， 卵泡闭锁和黄体溶解。 具体目标是：（A）确定 Fas抗原和Fas配体mRNA在肝癌细胞中的表达 用原位杂交技术观察了周期性小鼠卵巢的组织学变化。 量化变化 Fas抗原在诱导卵泡形成过程中的表达水平， 使用RT-PCR和流式细胞术检测黄体发育和退化。 (B)不同细胞因子对Fas抗原表达的影响 卵泡和黄体细胞分化和变性的介质 （促性腺激素、生长因子和细胞因子）。 这些药物对 测定细胞对Fas抗原介导的细胞凋亡的敏感性。 Fas抗原在人颗粒细胞/黄体细胞中表达， 受体与抗Fas抗原单克隆抗体的结合 (Fas mAB）诱导细胞凋亡。 这些结果提供了一个理由， 研究Fas抗原在触发卵泡发育中的潜在作用， 闭锁和/或黄体溶解。 卵巢卵泡受到刺激离开休眠期，开始 通过一种不确定的机制生长，从出生前开始并持续 在生活中。 卵泡对生长途径的承诺是 不可逆的，并导致两个可能的终点之一：排卵或 退化 卵巢中的卵泡少于1%， 注定要排卵，而绝大多数成为闭锁（退化）， 在发展过程中的一些状态。 卵泡排卵后 细胞分化并形成黄体。 黄体有一个 有限的寿命以退化告终。 启动的因素 对卵泡闭锁和黄体溶解知之甚少。 最近的证据 卵泡闭锁和黄体溶解是由细胞凋亡引起的， 深入了解这些过程。 对卵泡闭锁和黄体溶解的了解对于 允许开发更好的生育控制方法， 不孕症和提高生育能力。 了解那些 诱导卵巢细胞凋亡可能导致新的 卵巢癌和其他癌症的治疗。 此外，卵巢提供 一个优秀的和独特的模型，用于研究细胞凋亡，因为不断 细胞凋亡的周期在整个生命中发生在单个器官内。 这 实验室拥有卵巢生理学方面的设施和专业知识， 分子生物学来进行这些研究。

The overall goal is to identify triggering events and molecular mechanisms regulating programmed cell death of ovarian cells. The Fas antigen is a transmembrane receptor which triggers apoptosis (a form of programmed cell death). The project will define the role of the Fas antigen in ovarian follicular atresia and luteolysis. The specific aims are: (A) Determine Fas antigen and Fas ligand mRNA expression in specific cell types in the ovary of the cycling mouse using in situ hybridization. Quantify changes in levels of expression of Fas antigen during induction of follicular and corpus luteum development and regression using RT-PCR and flow cytometry. (B) Study modulation of expression of Fas antigen in vitro by various mediators of follicular and luteal cell differentiation and degeneration (gonadotropins, growth factors and cytokines). Effects of these agents on sensitivity of cells to Fas antigen-mediated apoptosis will be determined. The Fas antigen is expressed in human granulosa/luteal cells and engagement of the receptor with an anti-Fas antigen monoclonal antibody (Fas mAB) induces apoptosis. These results provide a rationale for studying a potential role of the Fas antigen in triggering follicular atresia and/or luteolysis. Ovarian follicles are stimulated to leave the resting stage and begin to grow by an undefined mechanism beginning before birth and continuing throughout life. Commitment of a follicle to the growth pathway is irreversible and results in one of two possible endpoints: ovulation or degeneration. Less than 1% of the follicles present in the ovary are destined to ovulate while the vast majority become atretic (degenerate) at some state in the developmental process. Following ovulation follicular cells differentiate and form the corpus luteum. The corpus luteum has a limited life-span terminating in degeneration. The factors initiating follicular atresia and luteolysis are poorly understood. Recent evidence that follicular atresia and luteolysis occur by apoptosis has provided new insight into these processes. An understanding of follicular atresia and luteolysis is essential to allow development of improved methods of fertility control, treatment of infertility and enhancement of fertility. Understanding pathways that induce apoptosis in ovarian cells may lead to development of novel therapies for ovarian and other cancers. Furthermore, the ovary provides an excellent and unique model for the study of apoptosis since continual cycles of apoptosis occur within a single organ throughout life. This laboratory has the facilities and expertise in ovarian physiology and molecular biology to undertake these studies.