The Molecular Biophysics and Tissue Biomechanics of Somite Morphogenesis

体节形态发生的分子生物物理学和组织生物力学

基本信息

批准号：
9896870
负责人：
SCOTT A HOLLEY
金额：
$ 38.36万
依托单位：
YALE UNIVERSITY
依托单位国家：
美国
项目类别：
财政年份：
2018
资助国家：
美国
起止时间：
2018-04-13 至 2022-03-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/9896870
关键词：
Adhesions Apical Automobile Driving Binding Proteins Biological Biological Assay Biology Biomechanics Biophysics Cadherins Cell Adhesion Cell Adhesion Molecules Cell Nucleus Cell Separation Cell Surface Proteins Cell model Cell-Cell Adhesion Cells Computer Models Computer Simulation Data Development Diffusion Disease Embryo Ensure Ephrins Epithelial Epithelial Cells Epithelium Exercise Extracellular Matrix Feedback Fibronectin Receptors Fibronectins Fluorescence Fluorescence Resonance Energy Transfer Genes Homeostasis ITGA5 gene Image Integrin alpha5beta1 Integrins Ligands Measures Mechanics Mediating Membrane Mesenchyme Mesoderm Modeling Molecular Molecular Conformation Morphogenesis Motion Movement Pathway interactions Protein Conformation Proteins Receptor Protein-Tyrosine Kinases Regulation Resources Role Side Signal Transduction Somites Spectrum Analysis Systems Analysis Techniques Testing Tissues Transgenic Organisms Vertebral column Zebrafish complement system epithelial to mesenchymal transition experimental study in silico in vivo kinematics laboratory experiment molecular dynamics molecular scale mutant protein protein interaction segregation spatiotemporal spine bone structure

项目摘要

Reciprocal regulation between the ECM and associated epithelial cells is integral to development, homeostasis and disease. Somites are segmental precursors of the vertebral column and musculature that form via a mesenchymal to epithelial transition. Somite morphogenesis is dependent upon a Fibronectin ECM, the Fibronectin receptor Integrin 51, the cell adhesion protein Cadherin 2 and bidirectional signaling via the receptor tyrosine kinase EphA4 and its membrane bound ligand Ephrin-B2a. These genes/pathways mediate cell-ECM adhesion, cell-cell adhesion and contact mediated cell repulsion, and our hypothesis is that the physical organizing activity of the somite boundary emerges via specific spatiotemporal intertwining of differential cell adhesion and ECM constrained cell repulsion. In Aim 1, fluorescence correlation spectroscopy (FCS) and fluorescence crosscorrelation spectroscopy (FCCS) will be used quantify protein diffusion and protein binding constants in vivo. These experiments will determine whether the segregation of these cell surface proteins occurs via diffusion and capture or active mobilization. Additionally, the roles of integrin 5, cadherin 2 and ephrin-b2a in driving these changes in subcellular localization will be elucidated by performing FCCS in live mutant embryos. In Aim 2, a systems analysis of cell motion will be used to quantify tissue biomechanics during somite morphogenesis in wild-type and mutant embryos. In Aim 3, we quantify the relative levels of Integrin activation via cytoplasmic signals versus via positive feedback through the ECM. Positive and negative feedback between biological mechanisms creates network effects that are hard to predict a priori and difficult to fully explore experimentally. in silico modeling will be used to systematically examine the relationships between cell adhesion, cell-ECM adhesion and cell contact mediated repulsion in somite morphogenesis in order to help interpret and prioritize more resource intensive wet-lab experiments.

ECM和相关上皮细胞之间的相互调节与发育，体内稳态和疾病。有时是椎骨的分段前体通过间质向上皮过渡形成的柱和肌肉。 somite 形态发生取决于纤连蛋白ECM，纤连蛋白受体整合素 51，细胞粘附蛋白钙粘蛋白2和通过接收器的双向信号传导酪氨酸激酶Epha4及其膜结合的配体Ephrin-B2A。这些基因/途径介导细胞ECM粘合剂，细胞细胞粘合剂和接触介导的细胞排斥，我们的假设是通过差分细胞粘合剂和ECM约束细胞的特定时空交织排斥。在AIM 1中，荧光相关光谱（FCS）和荧光互相关光谱（FCC）将用于量化蛋白质扩散和蛋白质结合常数在体内。这些实验将决定是否隔离这些细胞表面蛋白通过扩散和捕获或主动动员发生。此外，整联蛋白5，钙粘蛋白2和ephrin-b2a在推动这些变化中的作用通过在活突变胚胎中执行FCC来阐明亚细胞定位。在 AIM 2，将使用细胞运动的系统分析来量化组织生物力学期间的组织生物力学野生型和突变胚胎中的躯干形态发生。在AIM 3中，我们量化了亲戚通过细胞质信号激活整联蛋白的水平，而不是通过正面反馈 ECM。生物学机制之间的积极和负反馈产生网络效应这很难预测先验的，并且难以在实验上进行充分探索。在计算机建模中将用于系统地检查细胞粘合剂，细胞ECM之间的关系粘附和细胞接触介导的节点形态发生以帮助解释和优先考虑更多资源密集的湿与LAB实验。