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

胚胎发育依赖于多能细胞在空间和时间组织方式中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过程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重要，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以及细胞命运决定的内在与外在机制的相对贡献仍然缺乏特征。最近发展的基于成像和测序的谱系追踪技术现在提供了独特的机会，以前所未有的分辨率解决这个问题。我将在体内进行全基因组条形码，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生。然后，我将在鸡胚胎中使用高分辨率实时成像来跟踪实时展开的谱系轨迹，并获得对细胞行为的详细了解。我将整合这些数据，构建神经管谱系关系和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。通过将这种方法与标记未受干扰的邻近细胞的新方法相结合，我将研究系统的鲁棒性，以及细胞非自主机制对细胞命运的影响。总之，这将提供在分子上、空间上和时间上定义的亲缘关系的洞察力，为比较进化研究和发育障碍的调查奠定基础。

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.