Interrogating the Role of HSF1 in Ovarian Stress

项目摘要/摘要 人们早就知道，妊娠期的生理应激会导致压力和压力的改变 多代人的代谢反应。这样的信息必须通过生殖系传播； 然而，生理压力对生殖系的机械影响，更广泛地说，是否压力- 相关的变化发生在生殖细胞本身仍未被探索的水平上。母体的下丘脑- 垂体-肾上腺轴在妊娠应激时被激活，其下游效应是糖皮质激素，a 一类类固醇荷尔蒙，释放到血液中，然后与无处不在表达的 糖皮质激素受体(GR)。虽然已知应激诱导的糖皮质激素水平会损害卵母细胞 在成年女性中，其对胎儿卵母细胞的影响在很大程度上仍未被探索。有趣的是，GR有 已被证明可下调热休克因子1(HSF1)的活性。而HSF1的特征很好地体现在 除了它在蛋白质稳定中的作用外，它还被认为在减数分裂过程中调节卵母细胞的染色体结构。这 提案将调查卵母细胞固有的HSF1活性和母亲妊娠应激如何影响 胎儿卵母细胞发育。通过使用遗传小鼠模型，体内GR活性增加的模型，整体 卵巢清除、三维成像和定量分析，这项建议将(1)建立一个 HSF1在胎儿中的表达、定位和活动的全面三维时空图 卵巢和检测HSF1是否在胚胎减数分裂中发挥生殖细胞的内在作用，以及(2)确定两者之间的关系 胎儿卵母细胞中GR和HSF1之间的关系及其对胚胎减数分裂和卵母细胞生长的影响 和成熟。改善我们对全球增长带来的长期后果的有限理解 在发育中的生殖系上的活性是至关重要的，因为GR的有效药物激动剂地塞米松是 在怀孕期间常规使用，当早产令人担忧时，刺激肺成熟。 此外，阐明生理应激在卵母细胞中的机制效应将支持我们的 了解对暴露于压力风险较高的脆弱人群的后果- 诱发环境和社会经济条件。 为了成功地完成F31提案中概述的工作并实现我的职业目标，我选择了 为了在我的赞助人，加州大学的戴安娜·莱尔德博士的实验室里执行这项拟议的工作， 旧金山。我的共同赞助人Marco Conti博士和合作者Aditi博士的互补专业知识 Bhargava，以及F31奖学金提供的支持，保证我将接受技术培训和 在导师的指导下，我完成了博士前研究，并为生殖生物学领域做出了贡献。

PROJECT SUMMARY/ABSTRACT It has long been known that physiological stress during gestation can lead to the alteration of stress and metabolic responses over multiple generations. Such information must be transmitted through the germline; however, the mechanistic effects of physiological stress on the germline and, more broadly, whether stress- related changes occur at the level of the germ cells themselves remain unexplored. The maternal hypothalamic- pituitary-adrenal axis is activated upon gestational stress and its downstream effectors are glucocorticoids, a class of steroid hormones, which are released into the bloodstream then bind to the ubiquitously expressed Glucocorticoid Receptor (GR). While stress-induced levels of glucocorticoids are known to impair oocyte competence in adult women, their impact on the fetal oocyte remains largely unexplored. Interestingly, GR has been shown to downregulate the activity of Heat Shock Factor 1 (HSF1). While HSF1 is well characterized for its role in proteostasis, it is also known to regulate chromosome architecture in the oocyte during meiosis. This proposal will investigate how oocyte intrinsic HSF1 activity and maternal gestational stress influence fetal oocyte development. By using genetic mouse models, in vivo models of increased GR activity, whole ovary clearing, three-dimensional imaging, and quantitative analysis, this proposal will (1) establish a comprehensive, three-dimensional spatiotemporal map of HSF1 expression, localization, and activity in the fetal ovary and test whether HSF1 plays a germ cell-intrinsic role in embryonic meiosis and (2) identify the relationship between GR and Hsf1 in the fetal oocyte and the consequences for both embryonic meiosis and oocyte growth and maturation. Improving our limited understanding of the long-term consequences of global increased GR activity on the developing germline is crucial as the potent pharmacological agonist of GR, dexamethasone, is routinely administered during pregnancy to stimulate lung maturation when premature birth is a concern. Additionally, elucidating the mechanistic effects of physiological stress in the oocyte will support our understanding of the consequences for vulnerable populations who have a higher risk of exposure to stress- inducing environmental and socioeconomic conditions. To successfully complete the work outlined in this F31 proposal and to achieve my career goals, I have chosen to perform the proposed work in the laboratory of my Sponsor, Dr. Diana Laird, at the University of California, San Francisco. The complementary expertise of my Co-Sponsor, Dr. Marco Conti, and Collaborator, Dr. Aditi Bhargava, as well as the support provided by this F31 Fellowship assure I will receive the technical training and mentorship to complete my pre-doctoral research and contribute to the field of reproductive biology.