Molecular and Cellular Regulation of Uterine Morphogenesis

项目摘要 在哺乳动物中，子宫具有许多功能，包括精子的通道、胚胎植入和 胎儿怀孕。有流产史的妇女中有25%存在先天性子宫畸形 不孕不育。越来越多的人注意到子宫异常与生育同时存在。 疾病。因此，有必要进一步了解女性生殖发育，以更好地 管理和预防妇女的生殖疾病。女性生殖道由一对 上皮管被称为缪勒管(MD)。在胚胎发育过程中，苗勒管和 邻近的间质分化为输卵管、子宫、宫颈和阴道上部。在许多 对于哺乳动物，MD必须在中线融合才能正常的子宫形态发生。子宫的形成 MD首先分化、内陷、延长和融合到泌尿生殖窦(UGS)，然后 其次，在子宫形态发生过程中，两种MD在体中线相互融合。中的任何分歧 人类发育过程中的MD融合会导致子宫变异，这可能会阻碍健康的怀孕或 新生儿的分娩。虽然关于MD形成的研究很多，但目前还不清楚 基因和分子机制调控MD导管的融合。我们实验室的初步数据显示 Wnt7a基因敲除的小鼠由于未融合的MD导致子宫异常，这表明了一个重要的作用 Wnt7a在两个MD的融合中。目前，调节MD融合的细胞行为和机制 人们对此了解甚少。MD上皮与相邻间充质之间的串扰尚未被 在MD融合的背景下进行了研究。在目标1中，我将使用体外器官培养延时成像和 免疫荧光染色以确定哪些细胞变化与MD融合相协调。在《目标2》中，我将使用 用空间转录组学方法鉴定Wnt7a下游的差异表达基因 Wnt7a基因敲除胚胎。这项建议的目标是确定准确的时间、位置和长度 关于MD融合，在MD融合过程中发生了什么细胞变化，以及控制WNT7a的下游基因 MD融合对子宫形态发生的影响。我们的主要假设是WNT7a引导着 MD上皮与相邻的间质引导MD融合形成子宫的一部分。这个 Behringer实验室位于德克萨斯医学中心的MD Anderson，拥有生殖方面的专业知识 生物学、哺乳动物发育遗传学和小鼠遗传模型。在研究生培训期间，我将 与我的赞助商每两周一次会面，咨询委员会作为一个小组每两年会面一次，并与这一主题的专家会面 根据需要单独进行。我已经写了一章评论，并将写两本第一作者的出版物总结 我的发现。我将在我的研究生课程研讨会和两个全国性会议上展示我的工作。 每年一次。最终，这项工作将有助于我在中国进行博士后研究的长期目标 生殖科学和成为一名独立的生殖生物学研究人员。

Project Summary In mammals, the uterus serves many functions including a passage for spermatozoa, embryo implantation, and fetal gestation. Congenital uterine anomalies are present in 25% of women with a history of miscarriage and infertility. It has been increasingly noted that uterine anomalies are simultaneously present with reproductive diseases. For this reason, it is necessary to further understand female reproductive development to better manage and prevent reproductive diseases in women. The female reproductive tract develops from a pair of epithelial tubes called the Müllerian ducts (MD). During embryonic development, the Müllerian ducts and adjacent mesenchyme differentiate into the oviducts, uterus, cervix, and upper part of the vagina. In many mammals, the MD must fuse at the midline for proper uterine morphogenesis. Formation of the uterus occurs with the differentiation, invagination, elongation, and fusion first, of the MD to the urogenital sinus (UGS) and secondly, the two MD fuse to each other at the body midline during uterine morphogenesis. Any divergence in MD fusion during human development can lead to uterine variation that may prevent a healthy pregnancy or delivery of a newborn. While there have been many studies on MD formation, it is currently unknown which genes and molecular mechanisms regulate the fusion of the MD ducts. Preliminary data from our lab shows that Wnt7a knockout mice have uterine abnormalities as a result of unfused MD, suggesting an essential role for Wnt7a in fusion of the two MD. Currently, the cellular behaviors and mechanisms that regulate MD fusion are poorly understood. The crosstalk between MD epithelium with the adjacent mesenchyme has not been studied in the context of MD fusion. In Aim 1, I will use ex vivo organ culture time-lapse imaging and immunofluorescence staining to determine which cellular changes coordinate MD fusion. In Aim 2, I will use spatial transcriptomics to identify differentially expressed genes downstream of Wnt7a, using wild-type and Wnt7a knockout embryos. The objective of this proposal is to determine the precise timing, position, and length of MD fusion, what cellular changes occur during MD fusion, and the downstream genes of Wnt7a that govern MD fusion for uterine morphogenesis. Our primary hypothesis is that Wnt7a directs the crosstalk between MD epithelium with the adjacent mesenchyme to instruct MD fusion to form a portion the uterus. The Behringer laboratory located at MD Anderson in the Texas Medical Center has expertise in reproductive biology, mammalian developmental genetics, and mouse genetic models. During my graduate training, I will meet with my Sponsor biweekly, advisory committee as a group biannually, and with experts on the subject individually as needed. I have written a chapter review and will write two first-author publications summarizing my findings. I will present my work at both my graduate program seminars and at two national conferences annually. Ultimately this work will facilitate my long-term goals of conducting my postdoctoral studies in reproductive sciences and becoming an independent reproductive biology researcher.