Four-dimensional prediction and quantification of how physical forces impact organogenesis in zebrafish

物理力如何影响斑马鱼器官发生的四维预测和量化

• 批准号: 10271304
• 负责人: JEFFREY D AMACK
• 金额: $ 45.35万
• 依托单位: SYRACUSE UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2020
• 资助国家: 美国
• 起止时间: 2020-09-25 至 2025-08-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10271304
• 关键词: 3-Dimensional; Ablation; Actomyosin; Address; Affect; Anterior; Architecture; Biochemical; Biological Models; Biomechanics; Biophysical Process; Biophysics; Cell Shape; Cell model; Cells; Cellular biology; Complement; Complex; Congenital Abnormality; Defect; Developmental Biology; Developmental Process; Dorsal; Embryo; Embryonic Development; Environment; Epithelial; Extracellular Matrix; Formulation; Foundations; Four-dimensional; Goals; Health; Image; Image Analysis; Individual; Knowledge; Lasers; Lead; Left; Mathematics; Measurement; Mechanics; Methods; Mission; Modeling; Morphogenesis; Motion; Movement; Optics; Organ; Organ Model; Organogenesis; Output; Pattern; Phenotype; Physics; Play; Prevention; Public Health; Research; Resolution; Rod; Role; Shapes; Signal Transduction; Signaling Molecule; Structural Congenital Anomalies; Structure; Surface; Testing; Three-dimensional analysis; Time; Tissues; United States National Institutes of Health; Velocimetries; Vesicle; Work; Zebrafish; base; cell motility; convergent extension; disability; in vivo imaging; malformation; mathematical model; mechanical force; mechanical properties; morphogens; multidisciplinary; notochord; novel; particle; predictive modeling; prevent; programs; simulation; three dimensional structure

PROJECT SUMMARY/ABSTRACT Defects in programmed cell shape changes during embryonic development can disrupt organ morphogenesis and cause structural birth defects. There are fundamental gaps in our understanding of how cells change their shape during organ formation. While the biochemical signals and morphogen gradients that help govern organogenesis are well-studied, evidence is growing that robust control of organ form and function often also depends on multiple mechanical mechanisms that remain poorly understood. Thus, there is a critical need to tease apart how multiple mechanisms – including tissue-scale dynamic forces and cell-autonomous contractile forces – work together to generate “mechanical gradients” that program cell and organ shape during organ formation. A challenge is that mechanical perturbations that affect the entire embryo often result in the same global phenotype, making it difficult to pinpoint the role of each mechanism. Our long- term goal is to develop a combined cell biology and modeling toolkit that allows us to predict cell-scale phenotypes and appropriate perturbations that can be used to distinguish between multiple mechanical mechanisms. This project uses Kupffer’s vesicle (KV), a transient epithelial organ that establishes left-right asymmetry in the zebrafish embryo, as a model system. No upstream biochemical signaling gradients have been identified that regulate KV cell shapes as required for left-right patterning, but multiple mechanical mechanisms have been implicated. Preliminary results – from (4D = 3D + time) experimental perturbations and measurements of single KV cell shapes, and novel mathematical models that simulate interacting 3D tissue structures while retaining cell-scale resolution – lead us to formulate our central hypothesis that cell shape changes critical for KV organogenesis result from mechanical gradients generated by interactions between the KV and surrounding tissue structures as well as cell-autonomous contractile forces from inside KV. The goal of Aim 1 is to determine how interactions between KV and notochord impact cell shape changes. 4D modeling predictions for cell shapes and cell movement combined with live in vivo imaging and localized laser ablations will determine how asymmetric forces generated by the rod-like notochord impact KV cell shape changes during organogenesis. The goal of Aim 2 is to understand mechanisms by which actomyosin contractility in surrounding tailbud cells and inside KV generate KV cell shape changes. Novel mathematical models will predict how localized optical perturbations to tailbud mechanics, as well as perturbations to volume and cell- autonomous contractility in cells inside the KV, affect KV organ shape. Key outputs include a modeling toolkit for high-throughput simulations of dynamic interactions between complex 3D tissue structures complemented by a cell biology toolkit that tests model predictions with spatially and temporally modulated activation of biomechanical and biochemical signaling molecules. These results will pinpoint mechanical mechanisms that regulate organogenesis, and may ultimately aid in the prediction or prevention of birth defects.

项目摘要/摘要 胚胎发育过程中编程的细胞形状变化的缺陷会破坏器官形态发生 并导致结构性出生缺陷。 在器官形成期间的形状。 器官发生是充分研究的，有证据表明对器官形式和功能的强大控制通常是 取决于多种机械机制，这些机制仍然很差。 挑逗如何使用多种机制 - 包括组织尺度的动态力和细胞自治 收缩力 - 共同努力以产生程序单元和器官的“机械梯度” 在器官形成期间的形状是影响整个胚胎的机械扰动 通常会导致相同的全局表型，使其差异可以确定每个机制的作用。 术语目标是开发一个组合的细胞生物学和建模工具包，使我们能够预测细胞尺度 表型和适当的扰动，可以在多个多重机械之间存在 机制。该项目使用kupffer的囊泡（KV），这是一种瞬态上皮器官 斑马鱼胚胎中的不对称性作为模型系统。 被左右图案所需的调节KV细胞形状的质量化，但多个机械 机制已涉及初步结果 - （4D = 3D +时间）实验性 单个KV细胞形状的测量以及模拟相互作用3D组织的新型数学模型 结构同时保留细胞尺度的分辨率领导我们制定我们的中心假设，即细胞形状 对KV器官发生至关重要的变化是由于机械梯度在 KV和周围的组织结构以及内部KV的细胞自主收缩 AIM 1是确定KV和Notochord冲击细胞形状之间的相互作用 细胞形状和细胞运动的预测与活体内成像和局部激光消融结合 将确定杆状柱影响KV细胞形状的变化如何变化 在器官发生期间。 周围的尾bud细胞和KV内部会产生KV细胞形状的变化。 预测如何将光学扰动与尾串机械进行扰动，以及与体积和细胞的局面 KV内部细胞中的自主收缩性会影响KV器官形状。 用于对复杂的3D组织结构之间动态相互作用的高齿模拟。 通过细胞生物学工具包测试测试模型的预测，以空间和时间模块化 生物力学和生化信号分子。 调节器官发生，并最终可能有助于预测或投影出生缺陷。