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

斑马鱼脊髓稳健模式形成和形态发生的生物物理和遗传基础

基本信息

批准号：
9371529
负责人：
Tony Yu-Chen Tsai
金额：
$ 12.79万
依托单位：
HARVARD MEDICAL SCHOOL
依托单位国家：
美国
项目类别：
财政年份：
2017
资助国家：
美国
起止时间：
2017-09-30 至 2019-08-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/9371529
关键词：
Address Adhesions Affect Area Biological Assay Biophysics Cell Adhesion Cell Adhesion Molecules Cell Fate Control Cell Separation Cells Collaborations Complement Computer Simulation Controlled Study Development Developmental Biology Dorsal Down-Regulation E-Cadherin Embryo Embryonic Development Ensure Event Exhibits Gene Expression Gene Family Genes Genetic Genetic Engineering Genetic Screening Genetic Transcription Goals Grant Image Image Analysis Individual Institutes Interdisciplinary Study Knowledge Learning Measurement Measures Mechanics Mediating Mediator of activation protein Mentors Mentorship Methods Modeling Molecular Morphogenesis N-Cadherin Neural Tube Development Neural tube Optics Paper Participant Pattern Pattern Formation Phase Positioning Attribute Process Publications Publishing Regulation Regulator Genes Research Resolution Role Series Side Signal Transduction Snails Sorting - Cell Movement Specific qualifier value Spinal Cord Techniques Tissues Training Writing Zebrafish base biophysical techniques career cell determination cell fate specification cell motility cell type computer framework differential expression experience genetic approach graduate student in vivo migration mutant nerve stem cell neural patterning novel post-doctoral training programs skills spatiotemporal student mentoring success symposium three-dimensional modeling tool

项目摘要

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.

项目概要发育生物学中的一个重要问题是细胞命运规范和细胞运动如何发生在组织形态发生过程中进行协调，以确保所有细胞类型正确到达其所需位置。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。在这个建议，我将结合生物物理和遗传学方法来了解细胞命运规范和细胞如何斑马鱼神经管中的粘附是协调的。我早期博士后培训的研究表明确定 E-钙粘蛋白 (Cdh1)、N-钙粘蛋白 (Cdh2) 及其转录调节因子是神经祖细胞域的模式。基于这些初步发现，该提案旨在获得多从 Cdh1 介导的不同粘附力对脊髓模式形成的尺度理解和Cdh2在不同的神经祖细胞类型（Aim1）中，到控制空间的基因调控网络 Cdh1 和 Cdh2 表达模式 (Aim2)，到一个计算框架来模拟细胞分选组织规模（目标1）。目标1和目标2建立的实验平台将用于表征新颖的脊髓模式形成和形态发生过程中细胞粘附动力学的调节因子（目标 3）。该提案将结合我作为研究生培养的分析技能来分析时空细胞和亚细胞水平上粘附分子的动力学，在过程中获得的实验知识我早期的博士后培训是对斑马鱼进行基因改造并用单细胞对活胚胎进行成像决议，以及在 K99 指导阶段拟议的培训，以探索细胞力学和光学以高时空精度扰乱基因表达。 K99指导阶段的培训将整合我所有四位共同导师的专业知识，并将补充我过去的培训，形成一个完整的我自己的独立实验室的研究项目，用于测量、扰动和模拟时空动态脊髓发育过程中神经祖细胞中的粘附分子。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。通过与不同学术领域的所有四位共同导师学习和互动研究所，我将结合他们的力量来制定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研究项目，研究控制强健胚胎发育背后的细胞和亚细胞事件的时空动态。到目前为止我以研究经验的形式朝着这一目标取得了重大进展，成功出版物和合作的启动。然而，我坚信 K99 指导阶段会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我目前的博士后经验还缺乏其他方面的经验。