Identification of a Genetic Pathway Linking Temperature with Epigenetic Control of Gonad Determination in T. scripta

T. scripta 中温度与性腺决定的表观遗传控制之间的遗传途径的鉴定

• 批准号: 1854642
• 负责人: Blanche Capel
• 金额: $ 90万
• 依托单位: Duke University
• 依托单位国家: 美国
• 项目类别: Continuing Grant
• 财政年份: 2019
• 资助国家: 美国
• 起止时间: 2019-05-01 至 2024-04-30
• 项目状态: 已结题
• 来源: https://www.nsf.gov/awardsearch/showAward?AWD_ID=1854642&HistoricalAwards=false
• 关键词: Identification; Genetic; Pathway; Linking; Temperature

Temperature regulates male or female development in many reptiles, including the redeared slider turtle Trachemys scripta elegans, by controlling whether the embryonic gonad develops as a testis or an ovary. However, the mechanism through which temperature influences gonad development has been a puzzle since the discovery of this phenomenon 50 years ago. Previously, the Capel lab identified two genes that show strong expression differences in the turtle gonad at male- versus female-producing temperatures. In this project, the lab will determine how the expression of these genes is controlled by temperature, and how the activity of these genes controls testis versus ovary development. Results from this work will establish the mechanism underlying the effect of temperature on male or female development. The lab is actively involved in using this project as a platform to engage young students and the public in scientific research. This project is also a strong platform to advocate for evidence-based science in setting environmental policies to mitigate the effects of climate change, expected to strongly bias male/female ratios in species where temperature influences gonad development.The Capel lab previously showed that transcription of the epigenetic regulator KDM6B and the RNA-splicing factor, RBM20, are elevated in the turtle gonad at male-producing temperature. KDM6B activates the testis pathway by removing repressive histone marks at DMRT1, a conserved regulator of testis development. In this project, the lab plans to identify the molecular pathway linking temperature with changes in KDM6B and RBM20 transcription and splicing. The lab hypothesizes that a known repressor of KDM6B, pSTAT3, is differentially phosphorylated at high female-producing temperatures through activation of TRP channels, Ca2+ influx, and high activity of CAMK. To test this hypothesis, markers of the male and female pathways will be measured after knockdowns of TRP channels expressed in the gonad, after interference with Ca2+ signaling, or after of pSTAT3 inhibitors. Experiments will (1) determine whether RBM20 is also a target of pSTAT3, or downstream of KDM6B, and (2) test the role of RBM20 using a knock-down approach. Splicoform variants will be analyzed at the two temperatures using the lab's new T. scripta genome assembly. Results will establish the molecular basis for temperature-dependent gonad determination, and may uncover a conserved role for splicing in gonad development. The lab engages students in science research through creation of lesson plans that incorporate cutting edge research into local classrooms, and programs that encourage girls to pursue STEM careers. The lab also brings this science to public attention through podcast interviews, contribution to Duke's ongoing Art of the Scientist exhibition, and Duke's science cafe' series.This award reflects NSF's statutory mission and has been deemed worthy of support through evaluation using the Foundation's intellectual merit and broader impacts review criteria.

温度通过控制胚胎性腺是作为睾丸还是卵巢来控制许多爬行动物中的男性或女性发育，包括重建的滑块乌龟Trachemys Scripta秀丽隐杆线虫。但是，自50年前发现这种现象以来，温度影响性腺发育的机制一直是一个难题。以前，Capel Lab确定了两个基因，这些基因在雄性与产生女性的温度下显示出乌龟性腺中强烈的表达差异。在该项目中，实验室将确定这些基因的表达如何受温度控制，以及这些基因的活性如何控制睾丸与卵巢发育。这项工作的结果将确定温度对男性或女性发育作用的影响的基础机制。该实验室积极参与使用该项目作为与年轻学生和公众参与科学研究的平台。 This project is also a strong platform to advocate for evidence-based science in setting environmental policies to mitigate the effects of climate change, expected to strongly bias male/female ratios in species where temperature influences gonad development.The Capel lab previously showed that transcription of the epigenetic regulator KDM6B and the RNA-splicing factor, RBM20, are elevated in the turtle gonad at male-producing temperature. KDM6B通过在DMRT1（睾丸发育保守的调节剂）处删除DMRT1处的抑制性组蛋白标记来激活睾丸途径。在这个项目中，实验室计划确定将温度与KDM6B和RBM20转录和剪接的变化联系起来的分子途径。该实验室假设，通过激活TRP通道，Ca2+涌入和CAMK的高活性，在高女性产生温度下，已知的KDM6B PSTAT3的阻遏物在高女性产生温度下被差异化。为了检验该假设，在干扰Ca2+信号传导或PSTAT3抑制剂后，将测量男性和女性途径的标记。实验将（1）确定RBM20是否也是PSTAT3的目标，还是KDM6B的下游，以及（2）使用敲低方法测试RBM20的作用。将使用实验室新的T. scripta基因组组件在两个温度下分析剪接型变体。结果将建立依赖温度的性腺测定的分子基础，并可能发现在性腺发育中剪接的保守作用。该实验室通过制定将尖端研究纳入本地教室的课程计划，并鼓励女孩从事STEM职业的计划来参与科学研究。该实验室还通过播客访谈，对杜克大学持续的科学家展览的贡献以及杜克大学的科学咖啡馆系列的贡献，将这项科学引起公众的关注。该奖项反映了NSF的法定任务，并被认为是值得通过基金会的知识分子优点和更广泛影响的审查标准来通过评估来获得支持的。

项目成果

Temperature-dependent sex determination is mediated by pSTAT3 repression of Kdm6b

• DOI: 10.1126/science.aaz4165
• 发表时间: 2020-04
• 期刊: Science
• 影响因子: 56.9
• 作者: C. Weber;Yingjie Zhou;Jong Gwan Lee;L. Looger;Guoying Qian;C. Ge;B. Capel

The histone demethylase KDM6B regulates temperature-dependent sex determination in a turtle species

• DOI: 10.1126/science.aap8328
• 发表时间: 2018-05-11
• 期刊: SCIENCE
• 影响因子: 56.9
• 作者: Ge, Chutian;Ye, Jian;Capel, Blanche
• 通讯作者: Capel, Blanche

Sex determination without sex chromosomes

• DOI: 10.1098/rstb.2020.0109
• 发表时间: 2021-07
• 期刊: Philosophical Transactions of the Royal Society B
• 作者: C. Weber;B. Capel