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.

项目摘要/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众多单个单个细胞成连贯的高阶结构。虽然分子和遗传因素细胞器官的作用是这些因素调节的细胞生物学过程，例如细胞迁移，运动性和粘合剂的理解少得多，甚至对机械作用的了解更少在此过程中的力量。内胚层是最终引起的三个主要细菌层之一胃肠道和呼吸性上皮以及其他器官，例如甲状腺和胸腺。在他们期间发育，内皮细胞会在细胞表型中发生根本变化。在混血中，它们是高度迁移但最小的协调彼此，我们称为“单细胞迁移”的行为。最终但是，它们分化为执行重要屏障的相干组织中成熟的上皮细胞身体内部的功能。从动态的单细胞迁移到上皮纸的关键过渡期由于其光学透明度和对长期显微镜。在初步实验中，我们发现这个过渡期是通过由细胞内物理力驱动的质膜爆炸的变化，即膜到 - 皮层附着（MCA）和皮质张力。在此提案中，我们将对这些物理力并以两个具体目标确定其对内胚层发育的重要性。在第一个目的，我们将在体外和体内进行完整的方法，以定量确定是否以及什么范围MCA和收缩力在内胚细胞过渡过程中正在发生变化迁移到床单形成。在第二个目标中，我们将确定与Blebing相关的体内意义通过实验操纵散布频率在分化中专门用于内胚层开发的力斑马鱼胚胎中的内皮细胞。这项工作将促进我们对蜂窝作用的理解内胚层形态发生中的机械力，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。