Investigation of the mechanics of gastrulation in the chick embryo using new transgenic chicken lines

使用新的转基因鸡品系研究鸡胚原肠胚形成的机制

• 批准号: BB/T006781/1
• 负责人: Kees Weijer
• 金额: $ 79.69万
• 依托单位: University of Dundee
• 依托单位国家: 英国
• 项目类别: Research Grant
• 财政年份: 2020
• 资助国家: 英国
• 起止时间: 2020 至 无数据
• 项目状态: 未结题
• 来源: https://gtr.ukri.org/projects?ref=BB%2FT006781%2F1
• 关键词: Investigation; mechanics; gastrulation; chick; embryo

An important goal of the study of development of higher animals including humans is to understand how hundreds of thousands to millions of cells assemble into tissues that make organs and structures that make up an organism and which mechanisms control and integrate these behaviours. This coordination of cell behaviours is especially important at a critical early stage in embryonic development, gastrulation. During gastrulation the main body plan of the embryo is laid down and the main body axes emerge. Gastrulation involves large-scale, long-range cell movements and tissue deformations during which cells of the three body layers, the ectoderm, the mesoderm and endoderm take up their correct positions in the embryo. The endoderm is located inner most in the embryo forming the lining of the digestive tract and associated glands. It is surrounded by a middle layer the mesoderm that will make the muscles and the skeleton. The mesoderm is surrounded by the outmost layer, the ectoderm, which will form the epidermis (skin) and the brain and nervous system. Defects in the coordination of cell movements during gastrulation results in death of the embryo or in less severe cases in many major birth defects, such as the various forms of spina bifida and diverse heart defects.We study the mechanism of gastrulation in the chick embryo, that has the advantage that development takes place in the egg outside the mother animal and is therefore easily accessible to experimental observation and manipulation. The embryo is also flat and translucent which helps observation of cell movement during gastrulation. Gastrulation in chick embryos greatly resembles gastrulation in humans, which means that findings can be extrapolated to human development. We study how different cell behaviours such as cell division, shape changes, differentiation and motion are coordinated to generate these three embryonic tissues through specific chemical and mechanical cell-cell signalling mechanisms. Since these processes are highly dynamic we use time lapse microscopy to visualise these behaviours of the around 500.000 cells that make up the embryo of this stage of development. To do this we have developed and built a novel type of microscope, a light-sheet fluorescence microscope, that allows us to see almost all the cells in the embryo of a special chicken line in which the membranes of cells are marked with a green fluorescent protein. We have also developed extensive computational methods to extract quantitative information about relevant cell and tissue behaviours that drive gastrulation under normal and experimentally perturbed conditions. In this project we specifically test the hypotheses that forces produced by cell behaviours such as cell growth and movement are instrumental in the coordination of the complex cell behaviours of many cells in the developing embryo. Furthermore we expect that these forces feedback on the control of gene expression controlling the location and quantity of the various different types of cells in the embryo. To test this hypothesis we will generate new chicken lines that allow us to monitor and perturb the activity of critical elements of the actin-myosin cytoskeleton of cells that is responsible for generating these forces and correlating these with distinct cell and tissue behaviours using the above described live imaging and data analysis methodology.

研究较高动物在内的高等动物发展的一个重要目标是了解成千上万至数百万个细胞如何组装成构成有机体的器官和结构的组织，以及哪些机制控制和整合这些行为。细胞行为的这种协调在胚胎发育的关键早期阶段尤其重要。在胃肠道期间，放置了胚胎的主体计划，并出现主体轴。胃分解涉及大规模，远程细胞运动和组织变形，在此中，三个身体层的细胞，外胚层，中胚层和内胚层在胚胎中占据了正确的位置。内胚层位于胚胎中最内的，形成消化道和相关腺的衬里。它被中层包围，中胚层将使肌肉和骨骼形成。中胚层被最高层的外胚层包围，它将形成表皮（皮肤）以及大脑和神经系统。胃肠道过程中细胞运动配位的缺陷导致胚胎死亡或在许多主要的先天缺陷中，例如各种形式的脊柱裂和心脏缺陷的多种形式的病例。我们研究雏鸡胚胎中胃酸的机制，这具有在母亲动物之外的卵子，因此可以轻松地使用鸡蛋，因此可以轻松地使用母亲的卵子。胚胎也是扁平且半透明的，有助于观察胃部的细胞运动。雏鸡胚胎中的胃结构非常类似于人类的胃结构，这意味着发现可以推断到人类的发育中。我们研究了如何通过特定的化学和机械细胞 - 细胞信号传导机制来协调不同的细胞行为，例如细胞分裂，形状变化，分化和运动，以生成这三个胚胎组织。由于这些过程是高度动态的，因此我们使用延时显微镜可视化约500.000个细胞的这些行为，这些行为构成了这一发育阶段的胚胎。为此，我们开发并建立了一种新型的显微镜，即浅色荧光显微镜，使我们能够看到特殊鸡系的胚胎中几乎所有细胞，其中细胞的膜用绿色的荧光蛋白标记。我们还开发了广泛的计算方法，以提取有关相关细胞和组织行为的定量信息，这些细胞和组织行为在正常和实验扰动条件下驱动胃。在这个项目中，我们专门检验了细胞行为（例如细胞生长和运动）产生的力的假设，对发育中许多细胞在发育中的胚胎中许多细胞的复杂细胞行为的配位有助于。此外，我们希望这些力反馈控制控制胚胎中各种不同类型细胞的位置和数量的基因表达。为了检验这一假设，我们将生成新的鸡系，使我们能够监测和扰动细胞肌动蛋白膜肌球蛋白细胞骨架的关键元素的活性，这些元素负责产生这些力，并使用上述实时成像和数据分析方法将这些力与不同的细胞和组织行为相关联。

项目成果

The evolution of gastrulation morphologies.

• DOI: 10.1242/dev.200885
• 发表时间: 2023-04-01
• 期刊: Development (Cambridge, England)

A mechanochemical model recapitulates distinct vertebrate gastrulation modes.

• DOI: 10.1126/sciadv.adh8152
• 发表时间: 2023-12-08
• 期刊: SCIENCE ADVANCES
• 影响因子: 13.6
• 作者: Serra, Mattia;Najera, Guillermo Serrano;Chuai, Manli;Plum, Alex M.;Santhosh, Sreejith;Spandan, Vamsi;Weijer, Cornelis J.;Mahadevan, L.
• 通讯作者: Mahadevan, L.

Estimating stresses driving tissue flows using a stokes inverse problem

使用斯托克斯逆问题估计驱动组织流动的应力

• DOI: 10.1080/00207160.2022.2152281
• 发表时间: 2022
• 期刊: International Journal of Computer Mathematics
• 影响因子: 1.8
• 作者: Gao Y

Reconstruction of distinct vertebrate gastrulation modes via modulation of key cell behaviours in the chick embryo

• DOI: 10.1101/2021.10.03.462938
• 发表时间: 2021-10-03
• 期刊: bioRxiv
• 作者: Chuai, Manli;Serrano-Najere, G.;Weijer, C. J.
• 通讯作者: Weijer, C. J.