Determining the role of cellular forces in endoderm differentiation and development

确定细胞力在内胚层分化和发育中的作用

基本信息

批准号：
10527198
负责人：
Stephanie Woo
金额：
$ 18.31万
依托单位：
UNIVERSITY OF CALIFORNIA, MERCED
依托单位国家：
美国
项目类别：
财政年份：
2022
资助国家：
美国
起止时间：
2022-08-05 至 2024-07-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/10527198
关键词：
Actin-Binding Protein Actins Adhesions Adhesives Apoptosis Atomic Force Microscopy Behavior Biological Biological Assay Biological Process Bulla Cardiovascular system Cell Adhesion Cell Culture System Cell division Cell membrane Cell physiology Cells Cytoplasm Data Development Disease Dominant-Negative Mutation Embryo Embryonic Development Endocrine Glands Endoderm Endoderm Cell Epithelial Epithelial Cells Failure Frequencies Future Gene Expression Regulation Germ Layers Higher Order Chromatin Structure Hydrostatic Pressure Image In Vitro Individual Injections Maintenance Measurement Measures Mechanics Membrane Microscopy Molecular Genetics Morphogenesis Oils Optics Organ Organogenesis Phenotype Play Process Resolution Role Signal Transduction Site System Techniques Testing Thymus Gland Thyroid Gland Time Tissues Traction Force Microscopy Work Zebrafish airway epithelium base cell motility cell type craniofacial embryo tissue experimental study ezrin gastrointestinal gastrointestinal epithelium gastrulation in vivo mechanical force mutant novel optogenetics organ growth respiratory tool

项目摘要

PROJECT SUMMARY/ABSTRACT The formation of tissues and organs during embryonic development involves the carefully coordinated assembly of numerous individual cells into a coherent, higher-order structure. Although the molecular and genetic factors of organogenesis are well-studied, the cell biological processes that these factors regulate, such as cell migration, motility, and adhesion, are much less understood, and even less is known about role of mechanical forces in this process. The endoderm is one of the three primary germ layers that ultimately gives rise to the gastrointestinal and respiratory epithelia as well as other organs such as the thyroid and thymus. During their development, endodermal cells undergo a radical change in cellular phenotype. During gastrulation, they are highly migratory but minimally coordinated with each other, a behavior we term “single-cell migration.” Eventually however, they differentiate into mature epithelial cells within a coherent tissue that performs important barrier functions within the body. This critical transition period, from dynamic, single-cell migration to epithelial sheet formation, is especially accessible in the zebrafish embryo owing to its optical transparency and amenability to long-term microscopy. In preliminary experiments, we found that this transition period is accompanied by changes in plasma membrane blebbing that are driven by intracellular physical forces, namely membrane-to- cortex attachment (MCA) and cortical tension forces. In this proposal, we will quantitatively characterize these physical forces and determine their significance to endoderm development with two specific aims. In the first aim, we will take complementary in vitro and in vivo approaches to quantitively determine whether and to what extent MCA and contractile forces are changing during the transition of endodermal cells from single-cell migration to sheet formation. In the second aim, we will determine the in vivo significance of blebbing-associated forces specifically to endoderm development by experimentally manipulating blebbing frequency in differentiating endodermal cells within zebrafish embryos. This work will advance our understanding of the role of cellular mechanical forces in endodermal morphogenesis, and, importantly, establish a quantitative experimental pipeline that can be extended to other tissues and future studies.

项目概要/摘要胚胎发育过程中组织和器官的形成涉及精心协调的组装尽管分子和遗传因素将众多单个细胞整合成一个连贯的、高阶的结构。器官发生的相关因素已得到充分研究，这些因素调节的细胞生物学过程，例如细胞人们对迁移、运动和粘附的了解要少得多，对机械作用的了解更是少之又少。内胚层是最终产生的三个主要胚层之一。胃肠道和呼吸道上皮以及其他器官，例如甲状腺和胸腺。在发育过程中，内胚层细胞在原肠胚形成期间经历细胞表型的根本变化。高度迁移但彼此之间的协调程度最低，最终我们将这种行为称为“单细胞迁移”。然而，它们在具有重要屏障作用的连贯组织内分化成成熟的上皮细胞从动态的单细胞迁移到上皮层的关键过渡期。由于斑马鱼胚胎的光学透明性和易于形成，因此在斑马鱼胚胎中尤其容易接近在长期显微镜观察中，我们发现这个过渡期伴随着。由细胞内物理力（即膜对-）驱动的质膜起泡变化在本提案中，我们将定量描述这些皮质附着力（MCA）和皮质张力。物理力并确定它们对内胚层发育的重要性，有两个具体目标。目标，我们将采取互补的体外和体内方法来定量确定是否以及什么在内胚层细胞从单细胞转变过程中，MCA 和收缩力的变化程度在第二个目标中，我们将确定气泡相关的体内意义。通过实验操纵分化过程中的起泡频率来特异性地促进内胚层发育这项工作将增进我们对细胞作用的理解。内胚层形态发生中的机械力，并且重要的是，建立定量实验可以扩展到其他组织和未来研究的管道。