FAS AND BC1-2 AND GRANULOSA CELL APOPTOSIS

FAS 和 BC1-2 以及颗粒细胞凋亡

基本信息

批准号：
6164877
负责人：
NICHOLAS A CATALDO
金额：
$ 7.94万
依托单位：
STANFORD UNIVERSITY
依托单位国家：
美国
项目类别：
财政年份：
1996
资助国家：
美国
起止时间：
1996-03-01 至 2001-08-31
项目状态：
已结题

项目摘要

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.

候选人。候选人接受过妇产科培训，并且已完成生殖内分泌学专科研究金。他一直积极从事人类颗粒细胞的基础研究拥有超过 4 年的生理学经验，并拥有许多生理学方面的经验本提案中使用的技术。以第一作者发表论文五篇论文，另一篇已提交（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数量会增加，直到妊娠中期，之后持续下降直至更年期。没有被包围的卵母细胞颗粒细胞几乎在卵巢发育过程中经历闭锁，而卵泡在发育的每个阶段也会经历闭锁原始至第三窦期。绝大多数卵母细胞是因闭锁而丢失；只有少数成熟到排卵。在非人类模型中，窦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是受细胞外（细胞因子、生长因子）和细胞内机制。调节非细胞外因子人类颗粒细胞凋亡已有报道。拟议的研究寻求识别人类颗粒信号的细胞内途径细胞发生凋亡，以及凋亡的来源和身份细胞外效应子通过这些效应子激活细胞凋亡信号途径。更好地了解颗粒细胞的过程细胞凋亡可能有助于设计新疗法以延缓细胞凋亡伴随正常衰老和恶性肿瘤治疗的卵母细胞。该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 人类卵巢，以及细胞因子干扰素-γ (IFN- γ）和肿瘤坏死因子-α（TNF-α）在传输凋亡信号。这些研究也可能有助于确定是否凋亡信号起源于卵巢实质细胞或白细胞。据推测，在人颗粒细胞中，表达 Fas的激活促进细胞凋亡，而Bcl-2的表达过量其拮抗剂 Bax 可以防止细胞凋亡。进一步假设 IFN-γ 诱导 Fas 表达，而 TNF-α 可能诱导细胞凋亡通过其自身的受体。环境。医科大学生殖内分泌中心加利福尼亚州、旧金山市得到 NIH 拨款 #HD11979 的支持。包含在该中心有形态学、分子生物学和放射免疫分析核心，调查员可以使用其中的设施。小学赞助商的实验室和这些核心包含所有必要的设备对于所提出的研究。