Resolving Spatiotemporally-Specific Multicellular Dynamics In Vivo During Olfactory Neurogenesis

项目摘要 虽然已知许多神经缺陷和障碍是由特定发育障碍引起的， 细胞类型和/或与充分研究的信号传导途径有关，多个细胞的系统水平协调 在神经发生过程中，它们的信号输出的时空特异性仍然很差 明白这项建议的长期目标是利用方便快捷的 开发斑马鱼嗅上皮以定量表征细胞-细胞信号传导途径， 多细胞行为驱动脊椎动物中复杂神经元群体的组装。提出 实验将检验Notch和Wnt信号通路在空间和时间上提供 敏感的线索，引导干细胞迁移到嗅上皮和调节感觉神经发生。 将在亚细胞空间分辨率下完全在体内操作和分析小的细胞亚群， 具有亚分钟的时间分辨率，以便确定干细胞迁移的系统级协调， 在正常和中断的信号传导过程中的特异化和分化。首先，新工具和技术将 用于在嗅觉发育期间的特定时间和位置扰乱体内Wnt信号传导，并 算法模型和解释祖细胞的迁移行为。接下来，Notch信号将被操纵 以定量其对整个嗅觉系统神经发生的影响，并将鉴定靶基因， 是神经元特化和/或分化所需的。最后，转录因子insm 1a将被 类似地研究以确定其在Notch信号传导介导的嗅觉神经发生中的作用。的方法 将直接分析体内数据，以理解和预测多细胞行为，而不减少 生物复杂性，并帮助揭示表型，可能会促进我们对系统范围的广泛理解 神经元的分化和组装。

Project Summary While many neurological defects and disorders are known to result from developmental perturbations in specific cell types and/or are linked to well-studied signaling pathways, the system-level coordination of multiple cell populations and the spatiotemporal specificity of their signaling outputs during neurogenesis remains poorly understood. The long-term objective of this proposal is to take advantage of the accessible and rapidly developing zebrafish olfactory epithelium to quantitatively characterize cell-cell signaling pathways and multicellular behaviors that drive the assembly of complex neuronal populations in vertebrates. Proposed experiments will test the hypothesis that the Notch and Wnt signaling pathways provide spatially- and temporally- sensitive cues to guide stem cell migration into the olfactory epithelium and regulate sensory neurogenesis. Small subsets of cells will be manipulated and analyzed completely in vivo, at subcellular spatial resolution and with sub-minute temporal resolution, so as to determine the system-level coordination of stem cell migration, specification, and differentiation during both normal and disrupted signaling. First, new tools and techniques will be used to perturb Wnt signaling in vivo at specific times and locations during olfactory development and to algorithmically model and explain progenitor cells’ migratory behavior. Next, Notch signaling will be manipulated in vivo to quantitate its effects on olfactory system-wide neurogenesis, and target genes will be identified that are required for neuronal specification and/or differentiation. Finally, the transcription factor insm1a will be similarly investigated to determine its role in Notch signaling-mediated olfactory neurogenesis. The approaches in this proposal will directly analyze in vivo data to understand and predict multicellular behavior without reducing biological complexity and help uncover phenotypes that may advance our broad understanding of system-wide neuronal differentiation and assembly in vivo.