The biophysical and genetic basis of robust pattern formation and morphogenesis in zebrafish spinal cord

项目摘要 发育生物学中的一个重要问题是细胞命运的指定和细胞运动是如何进行的 在组织形态发生期间协调以确保所有细胞类型正确地到达其期望的位置。一 这种优雅协调的例子是斑马鱼脊髓中神经祖细胞的模式。在这 根据我的建议，我将结合联合收割机生物物理学和遗传学的方法来了解细胞命运的规范和细胞 在斑马鱼的神经管中，粘附是协调的。从我早期的博士后培训研究， 确定了E-钙粘蛋白（Cdh 1），N-钙粘蛋白（Cdh 2）及其转录调节因子作为 神经祖细胞结构域的模式化。在这些初步调查结果的基础上，这项建议旨在获得一个多方面的， 从Cdh 1介导的不同粘附力对脊髓模式形成的规模理解 和Cdh 2在不同的神经前体细胞类型（Aim 1），基因调控网络控制的空间 Cdh 1和Cdh 2表达模式（Aim 2），以计算框架模拟细胞分选， 组织标度（Aim 1）。目标1和2中建立的实验平台将用于表征新的 脊髓图案形成和形态发生过程中细胞粘附动力学的调节剂（目的3）。 这个建议将联合收割机结合我的分析能力，作为一个研究生，分析时空 粘附分子在细胞和亚细胞水平上的动力学， 我早期的博士后训练是对斑马鱼进行基因工程改造， 解决方案，以及在K99指导阶段提出的培训，以探测细胞力学和光学 以高时空精度干扰基因表达。K99辅导阶段的培训将 整合我的四位导师的专业知识，并将补充我过去的培训，形成一个完整的 在我自己的独立实验室的研究计划，以测量，扰动，和模型的时空动态 脊髓发育过程中神经前体细胞中的粘附分子。 我向独立过渡的计划包括我的四位共同导师的专业培训， 指导学生，管理实验室，撰写赠款，发表论文，在会议上展示研究成果，以及 建立科学合作。通过在不同的学术领域与所有四位共同导师学习和互动， 我将联合收割机结合他们的力量，形成我自己的实验室文化和指导风格。 我的长期职业目标是指导一个多学科的研究项目，研究控制 细胞和亚细胞事件的时空动态是胚胎发育的基础。到目前为止我 已经取得了重大进展，朝着这一目标的形式，研究经验，成功 出版物，并开始合作。然而，我坚信，K99指导阶段将有所帮助 通过提供额外的指导和培训，最大限度地提高我的成功机会， 从我目前的博士后经验来看，这是我所缺乏的。

PROJECT SUMMARY An important question in developmental biology is how cell fate specification and cell movement are coordinated during tissue morphogenesis to ensure all cell types reach their desired positions properly. One example of this elegant coordination is the patterning of neural progenitors in the zebrafish spinal cord. In this proposal, I will combine biophysical and genetic approaches to understand how cell fate specification and cell adhesion are coordinated in the zebrafish neural tube. Studies from my early postdoctoral training have identified E-cadherin (Cdh1), N-cadherin (Cdh2), and their transcriptional regulators as critical mediators for patterning of neural progenitor domains. Building on these initial findings, this proposal aims to obtain a multi- scale understanding of spinal cord pattern formation from the differential adhesion forces mediated by Cdh1 and Cdh2 in different neural progenitor cell types (Aim1), to the gene regulatory network controlling the spatial patterns of Cdh1 and Cdh2 expressions (Aim2), to a computational framework to simulate cell sorting at the tissue scale (Aim1). The experimental platform established in Aims 1 and 2 will be used to characterize novel regulators of cell adhesion dynamics during spinal cord pattern formation and morphogenesis (Aim 3). This proposal will combine my analytical skills developed as a graduate student to analyze spatiotemporal dynamics of adhesion molecules at cellular and subcellular levels, the experimental knowledge acquired during my early postdoctoral training to genetically engineer zebrafish and image live embryos with single cell resolution, and the proposed training during the K99 mentored phase to probe cell mechanics and optically perturb gene expression at high spatiotemporal precision. The training during the K99 mentored phase will integrate the expertise of all four of my co-mentors and will complement my past training to form a complete research program in my own independent lab to measure, perturb, and model spatiotemporal dynamics of adhesion molecules in neural progenitor cells during spinal cord development. My plan for transitioning to independence include professional trainings from all four of my co-mentors to mentor students, manage labs, write grants, publish papers, present research results in conferences, and establish scientific collaborations. By learning and interacting with all four co-mentors in different academic institutes, I will combine their strength to formulate my own lab culture and mentoring style. My long-term career goal is to direct a multidisciplinary research program studying the control of spatiotemporal dynamics of cellular and subcellular events underlying robust embryo development. So far I have achieved significant progress towards this goal in the form of research experience, successful publications, and initiation of collaborations. I firmly believe, however, that a K99 mentored phase will help maximize my chances for success by providing access to additional mentorship and training that would be otherwise lacking from my current postdoctoral experience.