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

• 批准号: 10196376
• 负责人: Jianping Fu
• 金额: $ 22.29万
• 依托单位: UNIVERSITY OF MICHIGAN AT ANN ARBOR
• 依托单位国家: 美国
• 财政年份: 2021
• 资助国家: 美国
• 起止时间: 2021-04-01 至 2023-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10196376
• 关键词: Adhesives; Alagille Syndrome; Anterior; Behavior; Biochemical; Biochemical Genetics; Biological Assay; Biological Clocks; Biological Pacemakers; Biomechanics; Biophysics; CADASIL; Cartilage; Cell Communication; Cell Density; Cell division; Cell physiology; Cells; Characteristics; Complex; Cues; Data; Defect; Dermis; Development; Developmental Process; Dimensions; Disease; Dissection; Embryo; Endothelium; Environment; Fertilization; Fibroblast Growth Factor; Fibronectins; Fingerprint; Glass; Goals; Hour; Human; Image; Impairment; In Vitro; Individual; Knowledge; Link; Malignant Neoplasms; Measurement; Mechanics; Mesoderm; Mesoderm Cell; Modeling; Molecular; Monitor; Mus; Paper; Pathway interactions; Pattern; Pattern Formation; Periodicity; Phenotype; Physiological; Physiological Processes; Play; Printing; Process; Regulation; Research; Role; Segmentation Clock Pathway; Signal Transduction; Skeletal Muscle; Sleeplessness; Somites; Source; Stretching; Surface; System; Tendon structure; Tissues; Vertebrates; Zebrafish; cell behavior; cell motility; developmental disease; experience; in vitro Assay; insight; live cell imaging; mechanical properties; monolayer; physical property; precursor cell; preservation; progenitor; response; scoliosis; somitogenesis; spatiotemporal; spine bone structure; stem cells; tool; vertebrate embryos

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.

项目摘要 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，心跳和体温发生。受损的生物振荡器会导致失眠为 癌症，对发展和分化有重大影响。细分时钟，生物学 从斑马鱼到人的振荡器保存良好，在调节周期体形成中起着关键作用 在脊椎动物的胚胎生成期间。虽然分割时钟的中央分子参与者有 长期以来已经确定，时钟嵌入了大型细胞内和细胞间网络中，以及如何响应 复杂的机械和生化微环境在很大程度上仍然未知。目标的目标 建议是开发一种体外测定，以实现复杂过程的定量解剖 参与斑马鱼的生物发生。前中胚层（PSM）细胞，参与参与的前体细胞 物质发生将从斑马鱼胚胎中分离出 具有可调机械提示的微电机械工具（底物刚度和机械拉伸） 生理范围。将进行实时成像以跟踪细胞行为，以监视其振荡 行为，细胞内信号传导活性和细胞力学（包括细胞骨架收缩力和细胞 刚度）是底物刚度和机械拉伸的函数。重要的是，我们的研究将进行 对于保留细胞 - 细胞通信的细胞菌落的背景。 我们提出的研究一起将导致有关机械和生化如何的新知识 微环境共同调节自我组织为发育模式的PSM细胞。