Interrogating the Role of HSF1 in Ovarian Stress

项目概要/摘要 人们早就知道，妊娠期间的生理应激会导致应激的改变， 多代人的代谢反应。这些信息必须通过生殖细胞传递; 然而，生理应激对生殖系的机械作用，以及更广泛地说，应激- 在生殖细胞本身的水平上发生的相关变化仍然未被探索。母亲的下丘脑- 垂体-肾上腺轴在妊娠应激时被激活，其下游效应物是糖皮质激素， 这类类固醇激素被释放到血液中，然后与普遍表达的 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， 尽管在成年女性中，它们对胎儿卵母细胞的影响在很大程度上尚未探索。有趣的是，GR 已显示下调热休克因子1（HSF 1）的活性。虽然HSF 1被很好地表征为 除了其在蛋白质稳定中的作用外，还已知其在减数分裂期间调节卵母细胞中的染色体结构。这 这项研究将探讨卵母细胞内在的HSF 1活性和母体妊娠应激如何影响 胎儿卵母细胞发育通过使用遗传小鼠模型，在GR活性增加的体内模型中， 卵巢透明化、三维成像、定量分析，本建议将（1）建立一个 胎儿HSF 1表达、定位和活性的全面三维时空图 卵巢和测试是否HSF 1发挥生殖细胞内在的作用，在胚胎减数分裂和（2）确定关系 胎儿卵母细胞中GR和Hsf 1之间的关系以及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GR增加的长期后果的有限理解 对发育中的生殖系的活性是至关重要的，因为GR的有效药理学激动剂地塞米松是 当担心早产时，在怀孕期间常规施用以刺激肺成熟。 此外，阐明卵母细胞中生理应激的机制效应将支持我们的研究。 了解易受压力风险较高的弱势群体的后果- 环境和社会经济条件。 为了成功完成F31计划书中概述的工作并实现我的职业目标，我选择了 在加州大学我的申办者Diana Laird博士的实验室进行拟定工作， 旧金山弗朗西斯科。我的共同申办者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， 导师完成我的博士前研究，并有助于生殖生物学领域。

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.