Regulation of neurodevelopmental cell behaviors in Ciona

Ciona 神经发育细胞行为的调节

• 批准号: 10701717
• 负责人: Christina Donata Cota
• 金额: $ 31.36万
• 依托单位: GEORGIA INSTITUTE OF TECHNOLOGY
• 依托单位国家: 美国
• 财政年份: 2022
• 资助国家: 美国
• 起止时间: 2022-09-12 至 2027-06-30
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10701717
• 关键词: Anatomy; Anterior; Automobile Driving; Axon; Behavior; Biochemical Pathway; Biological Availability; CRISPR/Cas technology; Cell Differentiation process; Cell Lineage; Cell Polarity; Cell physiology; Cells; Central Nervous System; Chordata; Cues; Development; Disease; Distal; Dorsal; Ectoderm; Embryo; Endosomes; Ensure; Epithelium; Fluorescence Microscopy; Gene Expression; Gene Expression Regulation; Genes; Genetic Transcription; Genomics; Homologous Gene; Human; Human Development; Individual; Invertebrates; Knowledge; Link; Malignant Neoplasms; Maps; Membrane; Molecular; Morphogenesis; Morphology; Movement; Nervous System; Neural Crest; Neural tube; Neurites; Neuronal Differentiation; Neurons; Organism; Outcome; Paraxial Mesoderm; Pathway interactions; Peripheral; Positioning Attribute; Process; Property; Proteins; Recycling; Regulation; Reproducibility; Research; Resolution; Role; Sensory; Shapes; Side; Specific qualifier value; Spinal Ganglia; Stereotyping; Study models; Synapses; Syndrome; System; Tail; Testing; Tight Junctions; Tissues; Transcriptional Regulation; Transforming Growth Factor beta; Urochordata; Vertebrates; Work; cell behavior; cell motility; cholinergic; connectome; extracellular; genetic architecture; genome editing; in vivo; in vivo imaging; innovation; insight; knockout gene; migration; nerve stem cell; neurodevelopment; notochord; polarized cell; programs; rab GTP-Binding Proteins; receptor; response; temporal measurement; therapy development; trafficking; transcription factor; transcriptomic profiling

PROJECT SUMMARY/ABSTRACT The regulation of transient cell behaviors and regulatory “states” is indispensable for the development of multicellular organisms. Knowledge of the genetic architecture and emergent properties of these processes is also key to developing therapies for congenital diseases and neurodevelopmental syndromes based on cellular reprogramming or genome editing. The objective of this proposal is to characterize the regulation and functions of potentially important genes that control polarized neuronal migration and axon outgrowth in the Ciona larval nervous system, which shares many anatomical and molecular features with the larger systems of their close relatives the vertebrates. The central hypothesis is that these processes are controlled by precise developmental regulation of genes encoding rate-limiting components of diverse biochemical pathways, which may vary according to developmental stage and neuronal subtype. The rationale underlying the proposed research is that, by exploiting the genomic and cellular simplicity afforded by invariantly developing Ciona embryos, one can study these processes in vivo, with greater spatial and temporal resolution. With only 231 neurons and a fully mapped “connectome”, the Ciona nervous system offers a singular opportunity to understand cell behaviors and developmental trajectories in a chordate nervous system at single-cell resolution. The central hypothesis will be tested by pursuing three specific aims: 1) Testing the role of instrinsic and extrinsic TGFβ pathway components in dynamically but invariantly polarizing neuronal progenitors; 2) Investigating the causal links between regulation of effectors of receptor trafficking and precisely timed inversion of intracellular polarity and axon outgrowth orientation. 3) Investigating the role of collective epithelial sheet-like migration in precise positioning and morphogenesis of differentiated neurons, and testing the involvement of tight junction proteins in regulating this unusual mode of collective migration. These aims will be pursued using an innovative approach that combines cell lineage-specific, CRISPR/Cas9-based somatic gene knockouts and live fluorescence microscopy. The expected outcomes of the proposed work include identifying previously unrecognized functions for conserved but poorly studied genes in neurodevelopment, and understanding how precise control over neuronal subtype-specific polarized cell behaviors can be achieved through transcriptional regulation of both intrinsic and extrinsic effector genes.

项目概要/摘要 瞬时细胞行为和调节“状态”的调节对于发育是必不可少的 的多细胞生物。了解这些基因的遗传结构和新兴特性 过程也是开发先天性疾病和神经发育疾病疗法的关键 基于细胞重编程或基因组编辑的综合症。该提案的目的是 表征控制极化神经元的潜在重要基因的调节和功能 海鞘幼虫神经系统中的迁移和轴突生长，该系统具有许多解剖学和 其近亲脊椎动物的更大系统的分子特征。中央 假设这些过程是由基因的精确发育调节控制的 编码不同生化途径的限速成分，这些成分可能会根据不同的情况而有所不同 发育阶段和神经元亚型。拟议研究的基本原理是，通过 利用不断发育的玻璃海鞘胚胎所提供的基因组和细胞的简单性，一种 可以以更高的空间和时间分辨率在体内研究这些过程。只有 231 个神经元 和一个完全映射的“连接组”，Ciona 神经系统提供了一个独特的机会 了解单细胞脊索动物神经系统的细胞行为和发育轨迹 解决。将通过追求三个具体目标来检验中心假设：1）检验 动态但不变极化神经元中的内在和外在 TGFβ 通路成分 祖先； 2) 研究受体运输效应器调节与受体运输之间的因果关系 细胞内极性和轴突生长方向的精确定时反转。 3）调查 集体上皮片状迁移在精确定位和形态发生中的作用 分化的神经元，并测试紧密连接蛋白在调节这种不寻常的过程中的参与 集体迁移的方式。将采用创新方法来实现这些目标，该方法结合了 细胞谱系特异性、基于 CRISPR/Cas9 的体细胞基因敲除和活体荧光显微镜。 拟议工作的预期成果包括确定以前未认识到的职能 神经发育中保守但研究很少的基因，并了解如何精确控制 神经元亚型特异性的极化细胞行为可以通过转录来实现 内在和外在效应基因的调节。