Quantitative characterization of a vertebrate segmentation clock response to biomechanical signals during zebrafish somitogenesis

斑马鱼体节发生过程中脊椎动物分段时钟对生物力学信号响应的定量表征

基本信息

批准号：
10369029
负责人：
Jianping Fu
金额：
$ 18.34万
依托单位：
UNIVERSITY OF MICHIGAN AT ANN ARBOR
依托单位国家：
美国
项目类别：
财政年份：
2021
资助国家：
美国
起止时间：
2021-04-01 至 2024-03-31
项目状态：
已结题

项目摘要

Project Summary Biological oscillators are essential to a variety of cellular, physiological and developmental processes, such as cell divisions, heartbeats, and somitogenesis. Impaired biological oscillators cause diseases from insomnia to cancer and have a significant impact on development and differentiation. Segmentation clock, a biological oscillator well-conserved from zebrafish to humans, plays a key role in regulating the periodic somite formation during vertebrate embryo somitogenesis. Although the central molecular players of the segmentation clock have been long identified, the clock is embedded in a large intra- and inter-cellular network, and how it responds to the complicated mechanical and biochemical microenvironments remains largely unknown. The goal of this proposal is to develop an in vitro assay that enables the quantitative dissection of the complex processes involved in the zebrafish somitogenesis. Presomitic mesoderm (PSM) cells, the precursor cells involved in the somitogenesis, will be isolated from zebrafish embryos and cultured and examined under an array of micromechanical tools with tunable mechanical cues (both substrate rigidity and mechanical stretching) across a physiological range. Live imaging will be conducted to track cell behaviors, to monitor their oscillatory behaviors, intracellular signaling activities and cell mechanics (including both cytoskeletal contractility and cell stiffness) as a function of substrate rigidity and mechanical stretching. Importantly, our studies will be conducted for both single cells as well as in the context of cell colonies where cell-cell communications are preserved. Together, our proposed studies will lead to new knowledge about how the mechanical and biochemical microenvironments jointly regulate PSM cells that self-organize into developmental patterns.

项目概要生物振荡器对于各种细胞、生理和发育过程至关重要，例如细胞分裂、心跳和体节发生。生物振荡器受损会导致失眠等疾病癌，并对发育和分化有重大影响。分段时钟，生物钟从斑马鱼到人类都保存完好的振荡器，在调节周期性体节形成中起着关键作用在脊椎动物胚胎体细胞发生过程中。尽管分段时钟的核心分子参与者人们很早就发现，时钟嵌入在一个大型的细胞内和细胞间网络中，以及它如何响应复杂的机械和生化微环境在很大程度上仍然未知。此举的目标建议开发一种体外测定方法，能够定量剖析复杂的过程参与斑马鱼的体细胞发生。前体中胚层 (PSM) 细胞，参与发育的前体细胞体节发生，将从斑马鱼胚胎中分离出来，并在一系列条件下进行培养和检查具有可调机械线索（基材刚性和机械拉伸）的微机械工具一个生理范围。将进行实时成像来跟踪细胞行为，监测其振荡行为、细胞内信号活动和细胞力学（包括细胞骨架收缩性和细胞刚度）作为基材刚度和机械拉伸的函数。重要的是，我们的研究将进行对于单个细胞以及保留细胞间通讯的细胞集落而言。总之，我们提出的研究将带来关于机械和生化如何进行的新知识。微环境共同调节 PSM 细胞自组织成发育模式。

项目成果

期刊论文数量（1）

专著数量（0）

科研奖励数量（0）

会议论文数量（0）

专利数量（0）

A human pluripotent stem cell-based somitogenesis model using microfluidics.

使用微流体的基于人类多能干细胞的体细胞发生模型。

DOI：
发表时间：
2023-11-01
期刊：
影响因子：
0
作者：
Liu, Yue;Kim, Yung Su;Xue, Xufeng;Kobayashi, Norio;Sun, Shiyu;Yang, Qiong;Pourquié, Olivier;Fu, Jianping
通讯作者：
Fu, Jianping