Role of the extracellular matrix during Wolffian/epididymal duct morphogenesis

细胞外基质在沃尔夫/附睾管形态发生过程中的作用

• 批准号: 10407029
• 负责人: Barry T. Hinton
• 金额: $ 32.84万
• 依托单位: UNIVERSITY OF VIRGINIA
• 依托单位国家: 美国
• 财政年份: 2018
• 资助国家: 美国
• 起止时间: 2018-08-01 至 2024-05-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10407029
• 关键词: Area; Atomic Force Microscopy; Attention; Behavior; Biomechanics; Cardiomyopathies; Cells; Clinical; Computer software; Congenital Abnormality; Deposition; Development; Duct (organ) structure; Elasticity; Embryo; Embryonic Development; Environment; Epididymis; Epithelial; Epithelial Cells; Event; Extracellular Matrix; Failure; Fetal Development; Genes; Goals; Growth; In Vitro; Lead; Liquid substance; Liver Fibrosis; Male Infertility; Mesenchymal; Mesenchyme; Microscopy; Modulus; Molecular; Morphogenesis; Movement; Mus; Nature; Neural Tube Development; Organ Culture Techniques; Outcome; Outcome Study; Pathologic; Play; Process; Property; Protein Tyrosine Kinase; Radial; Regulation; Reproductive Biology; Role; Sperm Maturation; Structure of mesonephric duct; Testing; Tissues; Tubular formation; cardiogenesis; cell motility; fetal; gastrulation; hearing impairment; human disease; intercalation; interest; male; male fertility; organ growth; real-time images; reproductive tract; second harmonic; tumor progression; two-photon

PROJECT SUMMARY The epididymis provides a unique luminal fluid microenvironment that allows for sperm maturation and survival, and disruptions to this function lead to male infertility. Further, disruptions to epididymal function may also arise as a consequence of abnormal fetal development, although very little is known either of the process of Wolffian duct/epididymal development or of the nature and causes of congenital defects that lead to male infertility. The central hypothesis of this application is that elongation and coiling of the Wolffian duct is crucial for the function of the resulting epididymis, and that failure to elongate and coil leads to male infertility. The overall goal is two-fold, to examine: (a) the mechanisms by which cells move to elongate the duct and (b) to dissect the underlying cellular and molecular mechanisms that regulate those cell movements. Although we have evidence that epithelial cells move by intercalating, we will test the hypothesis that intercalation-type movements of the mesenchyme cells that surround the duct and the en masse movement of mesenchymal cells in the interstitium contribute to ductal elongation and coiling. A combination of genetically modified mice, advanced microscopy including confocal, second harmonic two-photon, and atomic force microscopy, in vitro organ culture and contemporary software analyses will be used to test the hypotheses outlined in the following three specific aims: (1) To test the hypothesis that the stiffness (modulus) of the ECM undergoes dynamic changes during Wolffian duct morphogenesis thereby providing a biomechanical environment that promotes epithelial and mesenchymal cell intercalation and en masse movement of mesenchymal cells within the interstitium. (2) To test the hypothesis that radial intercalation of mesenchymal cells and the en mass movements of mesenchymal cells within the interstitium are major drivers of Wolffian duct elongation and coiling. (3) To test the hypothesis that Ptk7 and Rac1 regulate radial intercalation and en masse movement of mesenchymal cells via regulating ECM biomechanical properties through changes in its deposition and assembly during Wolffian duct development, which in turn are important for male fertility. Therefore, this application focuses on the role of the ECM during Wolffian duct morphogenesis paying special attention to the importance of its assembly, distribution and stiffness, and how this is coordinated to regulate mediolateral and radial intercalation of epithelial and mesenchymal cells respectively, and the en masse movement of mesenchymal cells within the interstitium. Coordination of these events is critical for Wolffian/epididymal duct development, and therefore, male fertility. The anticipated outcomes of this study will not only have a major impact on an area of reproductive biology that has been poorly understood, but will also contribute to our understanding of the fundamental process of tubular morphogenesis. Specifically they will provide an understanding as to how the regulation of growth of the epididymis during development is important clinically.

项目摘要 附睾提供了独特的腔液微环境，可实现精子成熟和存活， 对此功能的中断导致男性不育症。此外，可能会出现对附睾功能的破坏 由于胎儿发育异常，尽管沃尔夫人的过程很少 导管/附睾发育或先天性缺陷导致男性不育症的性质和原因。这 该应用的中心假设是沃尔夫管的伸长和盘绕对于 由此产生的附睾的功能，并且无法拉长和线圈导致男性不育症。 总体目标是两个方面，要检查：（a）细胞移动以拉长管道的机制和（b） 剖析调节这些细胞运动的潜在细胞和分子机制。虽然我们 有证据表明上皮细胞通过插入而移动，我们将检验以下假设。 围绕管道和间充质运动的间质细胞的运动 间质中的细胞有助于导管伸长和盘绕。转基因小鼠的组合， 晚期显微镜，包括共聚焦，第二谐波两光子和原子力显微镜，体外 器官文化和现代软件分析将用于测试以下概述的假设 三个具体目的：（1）测试以下假设：ECM的刚度（模量）经历动态 沃尔夫（Wolffian）管道形态发生期间的变化，从而提供了促进的生物力学环境 上皮和间质细胞的插入以及间充质细胞在 间质。 （2）测试了间充质细胞和EN质量的径向插入的假设 间充质细胞在间质中的运动是沃尔夫管道伸长的主要驱动因素， 盘绕。 （3）检验PTK7和RAC1调节径向插入的假设 间充质细胞通过调节ECM生物力学特性通过其沉积和 沃尔夫（Wolffian）管道发育过程中的组装，进而对男性生育至关重要。因此，这个 应用的重点是ECM在沃尔夫语导管形态发生过程中的作用，特别关注 其组装，分布和刚度的重要性，以及如何协调以调节中外侧和 上皮细胞和间质细胞的径向插入，以及大体上的运动 间充质细胞中的间质细胞。这些事件的协调对于沃尔夫语/附子管至关重要 发展，因此是男性生育能力。这项研究的预期结果不仅将有一个重大 对繁殖生物学领域的影响，该领域知之甚少，但也会为我们的 了解管状形态发生的基本过程。特别是他们将提供 了解在发育过程中附睾增长的调节如何在临床上很重要。