FAS AND BC1-2 AND GRANULOSA CELL APOPTOSIS

候选人。应聘者接受过产科和妇科方面的培训， 已完成生殖内分泌学的专科奖学金。他 一直积极从事人类颗粒细胞的基础研究 有超过4年的生理学经验，并有多项 在这项建议中要使用的技术。他已经出版了五本第一作者 提交了一份文件，另一份已提交(附录)。 研究计划。在胚胎和胎儿发育过程中， 人类卵巢中的卵母细胞增加到妊娠中期，之后 在更年期之前一直在稳步下降。没有被包围的卵子 接近卵巢发育的颗粒细胞经历闭锁，而 卵泡在发育的每个阶段都会经历闭锁。 原始期至第三胃窦期。绝大多数的卵母细胞是 由于闭锁而丢失；只有少数成熟到排卵。在非人类中 模型中，窦状卵泡闭锁与颗粒细胞有关 细胞凋亡，或程序性细胞死亡。细胞凋亡的激活可以 受细胞外(细胞因子，生长因子)和 细胞内机制。调节非细胞因子的细胞外因子 已有人颗粒细胞凋亡的报道。建议进行的研究 寻求确定向人类颗粒发出信号的细胞内通路 细胞发生凋亡，以及来源和身份 细胞外效应器通过这些激活细胞凋亡信号 小路。更好地理解颗粒细胞的发育过程 细胞凋亡可能有助于设计新的治疗方法来延缓 伴随着正常衰老和治疗恶性肿瘤的卵母细胞。 这项提案的具体目的是调查 细胞内凋亡调节分子Fas/APO-1和Bcl-2在细胞内的表达 人卵巢，以及细胞因子干扰素-γ的作用。 )和肿瘤坏死因子-α(TNF-α)在传递 细胞凋亡信号。这些研究也可能有助于确定 凋亡信号起源于卵巢实质细胞或来自 白细胞。假设在人类颗粒细胞中，表达 Fas活化促进细胞凋亡，而Bcl2过度表达 其拮抗剂BAX可防止细胞凋亡。进一步假设 干扰素-γ诱导Fas表达，而肿瘤坏死因子-α可诱导细胞凋亡 通过它自己的受体。 环境加州大学生殖内分泌学中心 加利福尼亚州、旧金山由美国国立卫生研究院拨款#HD11979资助。包括在 这些中心是形态学、分子生物学和放射免疫分析核心， 其设施可供调查员使用。初选 赞助商的实验室，这些核心包含所有必要的设备 对于提议的研究。

Candidate. The candidate has been trained in obstetrics and gynecology and has completed a subspecialty fellowship in Reproductive Endocrinology. He has been actively engaged in basic research on human granulosa cell physiology for over 4 years and has experience with a number of the techniques to be used in this proposal. He has published five first-author papers, and another has been submitted (Appendix). Research plan. During embryonic and fetal development, the number of oocytes in the human ovary increases until mid-gestation, after which it steadily declines until the menopause. Oocytes which are not surrounded by granulosa cells nearly i ovarian development undergo atresia, while follicles also undergo atresia at every stage of development from the primordial to the tertiary antral stage. The vast majority of oocytes are lost through atresia; only a minority mature to ovulation. In non-human models, atresia of antral follicles has been associated with granulosa cell apoptosis, or programmed cell death. Activation of apoptosis can be regulated by both extracellular (cytokine, growth facto) and intracellular mechanisms. The extracellular factors which regulate non- human granulosa cell apoptosis are being reported. The proposed studies seek to identify the intracellular pathways that signal the human granulosa cell to undergo apoptosis, as well as the source and identity of th extracellular effectors that activate the apoptotic signal through these pathways. A better understanding of the process of granulosa cell apoptosis may aid in the design of new therapies to retard the loss of oocytes that accompanies both normal aging and therapies for malignancy. The specific aims of this proposal are to investigate the roles of the intracellular apoptosis-regulating molecules Fas/APO-1 and Bcl-2 in the human ovary, as well as the roles of the cytokines interferon-gamma (IFN- gamma) and tumor necrosis factor-alpha (TNF-alpha) in transmitting the apoptotic signal. These studies may also help to determine whether the apoptotic signal originates within ovarian parenchymal cells or from leukocytes. It is hypothesized that in human granulosa cells, expression and activation of Fas promotes apoptosis, while Bcl-2 expression in excess of its antagonist Bax prevents apoptosis. It is further hypothesized that IFN-gamma induces Fas expression, while TNF-alpha may induce apoptosis through its own receptor. Environment. The Reproductive Endocrinology Center at the university of California, San Francisco is supported by NIH Grant #HD11979. Included in the Center are Morphology, Molecular Biology, and Radioimmunoassay Cores, the facilities of which are available to the investigator. The Primary Sponsor's laboratory and these Cores contain all of the necessary equipment for the studies proposed.