Mechanisms of ovarian endocrine disruption at single-cell resolution

单细胞分辨率下卵巢内分泌紊乱的机制 内分泌干扰物(EDCs)是干扰内源性激素的外源性化学物质 合成、代谢和信号传递。在性腺发育早期接触EDC被怀疑起到 在晚年卵巢发育不全中的作用。在分子和分子水平上促进对内分泌干扰的理解 细胞水平对于确定成人型生殖疾病可能的发育起源至关重要。 这需要具有短寿命的灵敏活体模型和创新的分析工具来检测基因 由化学物质暴露引起的干扰。日本青花鱼是世界上最大的鱼之一。 研究了用于EDC筛查的发育和生殖影响的实验模型。不仅是因为 与其他鱼类模型相比，青竹基因组有广泛的注释，但它有共同的染色体性别 测定，激素受体序列同源性，性激素信号轴与人类。 卵巢发育期间暴露在雌激素污染的水中会导致卵母细胞显著延迟 青竹成虫发育成熟，产卵量减少。然而，通过哪些具体机制 发育中的卵巢的异种细胞群对雌激素类化学物质的反应尚不清楚。 这项建议旨在调查早期发育阶段暴露于外源雌激素如何改变基因调控。 卵巢细胞分化中的网络，以及长期转录变化与之相关 女性生育力下降。我的工作假设是，生命早期接触雌激素滴滴涕 在卵巢体细胞性腺细胞中诱导长期的转录效应，降低以后的生育能力。我 将通过将青竹暴露于与环境相关的o，p‘-水平来研究这一假设 二氯二乙基三氯乙烷(o，p‘-DDT)，是一种遗留有机氯农药的雌激素异构体， 仍然是一个持续的公共卫生问题。暴露将发生在卵巢发育的关键时期 在功能、细胞和转录水平上定义组织效应。 目标1将确定o，p‘-DDT暴露的长期生殖后果 卵巢分化的功能繁殖评估和雌激素的靶向量化--和 类固醇生成相关基因。AIM 2将使用单细胞转录(scRNA-seq)技术获得 全面了解哪些卵巢体细胞类型和基因调控网络(包括类固醇激素和 依赖和激素非依赖)，会被早期接触o，p‘-DDT永久改变。 拟议的工作将阐明外源性雌激素在体内引起的持久的功能和分子变化。 青竹的卵巢分化和卵泡组装与胎儿卵巢发育相一致 人类。对DDT诱导基因的综合鉴定可能会揭示新的暴露生物标志物和 内分泌和生殖疾病的可能机制。

Mechanisms of Ovarian Endocrine Disruption at Single-Cell Resolution Endocrine disrupting chemicals (EDCs) are exogenous chemicals that interfere with endogenous hormone synthesis, metabolism and signaling. EDC exposure during early gonadal development is suspected to play a role in ovarian dysgenesis later in life. Advancing understanding of endocrine disruption at the molecular and cellular level is essential for determining possible developmental origins of adult-onset reproductive disease. This demands sensitive in vivo models with short lifespans and innovative analytical tools to detect genetic perturbations induced by chemical exposures. The Japanese medaka fish (Oryzias latipes) is among the most studied experimental models used in EDC screening for developmental and reproductive effects. Not only is the medaka genome extensively annotated compared to other fish models, but it shares chromosomal sex determination, hormone receptor sequence homology, and sex hormone signaling axes with humans. Exposure to estrogen-contaminated water during ovarian development causes significantly delayed oocyte maturation and reduced egg production in adult medaka. However, specific mechanisms by which heterogenous cell populations of the developing ovary respond to estrogenic chemicals remain unclear. This proposal seeks to investigate how early developmental exposures to xenoestrogens alter gene regulation networks in differentiating ovarian cells, and which long-term transcriptional changes are associated with decreased female fertility. My working hypothesis is that early life stage exposures to estrogenic DDTs induce long-term transcriptional effects in ovarian somatic gonad cells, reducing fertility later in life. I will investigate this hypothesis by exposing medaka to environmentally relevant levels of o,p’- dichlorodiethyltrichloroethane (o,p’-DDT), an estrogenic isomer of the legacy organochlorine pesticide that remains a persistent public health concern. Exposures will occur during a key window of ovarian development to define organizational effects at the functional, cellular, and transcriptional levels. Aim 1 will define the long-term reproductive consequences of o,p’-DDT exposures during an early window of ovarian differentiation using functional breeding assessments and targeted quantification of estrogen- and steroidogenesis-related genes. Aim 2 will use single cell transcriptomics (scRNA-seq) to gain a comprehensive view of which ovarian somatic cell types and gene regulatory networks (both steroid hormone- dependent and hormone-independent), are permanently altered by early stage o,p’-DDT exposure. The proposed work will elucidate lasting functional and molecular changes induced by xenoestrogens during ovarian differentiation and follicular assembly in medaka, which aligns with fetal ovarian development in humans. Comprehensive identification of DDT-inducible genes may reveal novel exposure biomarkers and plausible mechanisms of endocrine and reproductive diseases.