Multidimensional investigation of cellular dynamics and lineage relationships in the vertebrate neural tube

脊椎动物神经管细胞动力学和谱系关系的多维研究

• 批准号: EP/X031225/1
• 负责人: Giulia Boezio
• 金额: $ 25.97万
• 依托单位: The Francis Crick Institute
• 依托单位国家: 英国
• 项目类别: Fellowship
• 财政年份: 2023
• 资助国家: 英国
• 起止时间: 2023 至 无数据
• 项目状态: 未结题
• 来源: https://gtr.ukri.org/projects?ref=EP%2FX031225%2F1
• 关键词: Multidimensional; investigation; cellular; dynamics; lineage

Embryonic development relies on multipotent cells producing distinct cell types in a spatially and temporally organised manner.Understanding cellular lineage - the history and relationships between cells in a tissue - establishes a framework to explaindevelopment and to explore the interplay between extrinsic signals and intrinsic competence in the generation of cellular diversity.This is particularly relevant in the vertebrate spinal cord, where recent analyses have revealed even greater molecular andorganisational complexity than previously appreciated. However, lineage relationships between different cell types in the spinal cordremain poorly characterized, as well as the relative contribution of intrinsic versus extrinsic mechanisms to cell fate decisions. Recentlydeveloped imaging- and sequencing-based lineage tracing techniques now offer the unique opportunity to address this question atan unprecedented resolution. I will perform in vivo whole-genome barcoding followed by single-cell transcriptomics to characterizethe ontogeny of neurons and glial cells. I will then use high-resolution live imaging in chick embryos to track lineage trajectoriesunfolding in real-time and obtain a detailed understanding of cellular behaviour. I will integrate these data to build a comprehensivemap of lineage relationships and cell identity acquisition in the neural tube. Moreover, these approaches are compatible withsimultaneous molecular perturbations, allowing me to test the plasticity of progenitors' potential. By combining this approach with anovel method to label unperturbed neighbouring cells, I will investigate the robustness of the system, as well as the effect of cell nonautonomousmechanisms on cell fate. Together, this will provide insight into molecularly, spatially, and temporally defined in vivolineage relationships, establishing a foundation for comparative evolutionary studies and the investigation of developmentaldisorders.

胚胎发育依赖于多能细胞以空间和时间上有组织的方式产生不同的细胞类型。了解细胞谱系-组织中细胞之间的历史和关系-建立了一个框架来解释发育和探索细胞多样性产生中外在信号和内在能力之间的相互作用。这在脊椎动物脊髓中尤其相关，其中最近的分析揭示了比以前认识到的更大的分子和组织复杂性。然而，脊髓中不同细胞类型之间的谱系关系特征不佳，以及内在与外在机制对细胞命运决定的相对贡献。最近开发的成像和测序为基础的谱系追踪技术，现在提供了独特的机会，以解决这个问题，在一个前所未有的决议。我将在体内进行全基因组条形码编码，然后进行单细胞转录组学，以表征神经元和神经胶质细胞的个体发育。然后，我将在鸡胚中使用高分辨率的实时成像来实时跟踪谱系发育，并获得对细胞行为的详细了解。我将整合这些数据来建立一个谱系关系和神经管中细胞身份获取的综合地图。此外，这些方法与同时发生的分子扰动兼容，使我能够测试祖细胞潜力的可塑性。通过结合这种方法与anovel方法来标记未受干扰的相邻细胞，我将研究系统的鲁棒性，以及细胞的non-bondyousmechanisms对细胞命运的影响。总之，这将提供深入了解分子，空间和时间上定义的vivolineage关系，建立比较进化研究和发育障碍的调查的基础。