How temporal transcription factors regulate guidance cues and dendrite morphogenesis in Drosophila somatosensory circuits

时间转录因子如何调节果蝇体感回路中的引导线索和树突形态发生

• 批准号: 10609392
• 负责人: Jake Henderson
• 金额: $ 4.75万
• 依托单位: UNIVERSITY OF CHICAGO
• 依托单位国家: 美国
• 财政年份: 2022
• 资助国家: 美国
• 起止时间: 2022-04-01 至 2025-03-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10609392
• 关键词: 3-Dimensional; Adopted; Affect; Afferent Neurons; Animals; Architecture; Area; Axon; Biological Assay; Birth Order; Confocal Microscopy; Cues; Data; Daughter; Dendrites; Development; Disease; Disparate; Drosophila genus; Drosophila melanogaster; Environment; Family; Gene Expression; Injury; Interneurons; Knowledge; Label; Lateral; Ligands; Location; Maps; Modeling; Molecular; Morphogenesis; Morphology; Nerve; Nervous System; Neurons; Pain; Pathology; Peripheral Nervous System; Positioning Attribute; Process; Proprioceptor; Receptor Gene; Receptor Signaling; Regulation; Role; Rosaniline Dyes; Signaling Molecule; Specific qualifier value; Synapses; System; Temperature; Testing; Tissue-Specific Gene Expression; Touch sensation; Work; axon guidance; cell type; confocal imaging; differential expression; genetic manipulation; innovation; loss of function; nerve stem cell; neuroblast; neurodevelopment; neuron development; neuronal circuitry; postmitotic; receptor; receptor expression; self-renewal; single-cell RNA sequencing; somatosensory; stem cells; therapy development; tool; transcription factor

ABSTRACT Properly developed somatosensory circuits are critical for any animal to sense and interact with their environment. The development of somatosensory circuits requires that each neuron within the circuit to correctly position its axons and dendrites. This allows them to establish specific synaptic connections and for circuits to assemble. Within somatosensory circuits, interneurons represent are the most numerous neuron type. The other types are sensory neurons of the peripheral nervous system. In comparison to sensory neurons, interneurons are under investigated due to lack of tools and knowledge about somatosensory circuit architecture. Molecular factors, like temporal transcription factors and guidance cues, are known to regulate different aspects of neuronal development, however the role of these molecular factors has not been deeply investigated in interneuron dendrite development, synaptic partner selection, and circuit formation. I will test the central hypothesis that temporal transcription factors regulate guidance receptor expression, and this establishes specific dendritic morphologies and synaptic connections. To investigate this central hypothesis, in the Drosophila nerve cord, I will be using the neuroblast 3-3 stem cell and the EL interneurons it generates as a model. In Aim 1, I will test the working hypothesis in NB3-3, temporal transcription factors, Cas and Nab, activate different sets of guidance cue receptors in its early-born and late-born EL daughter interneurons, and that this determines differential dendrite positioning. I use single cell RNA-sequencing, NB3-3 specific gene manipulation of Cas and Nab, and confocal microscopy to examine dendrite morphology. I expect to connect the role of temporal transcription factors to the role of guidance cue receptor expression. Additionally, I expect to introduce an innovative sequencing approach to our current analysis of somatosensory circuit development. In Aim 2, I will test the working hypothesis that the guidance receptor Roundabout3 (Robo3) is necessary in early-born EL dendrites and sufficient in late-born EL dendrites to regulate dendrite morphogenesis and position. I will perform Robo3 subcellular localization assays alongside gain and loss-of-function assays of Robo3 to examine the function of Robo3 in EL dendrites. This aim will serve to identify the requirement of Robo3 in interneurons and develop new assays to analyze Robo3 localization. Overall, this proposal will connect to previous disparate areas of somatosensory circuit development. This work will also enable future research to investigate the broad conservation of neuronal circuit specification and development of therapies for neurodevelopmental pathologies in which circuit wiring is disrupted.

抽象的 正确开发的体感电路对于任何动物都至关重要 环境。体感电路的发展要求电路中的每个神经元正确 放置其轴突和树突。这使他们能够建立特定的突触连接，并使电路 集合。在体感电路中，中间神经元代表是最多的神经元类型。另一个 类型是周围神经系统的感觉神经元。与感觉神经元相比 由于缺乏有关体感应电路架构的工具和知识，因此未经研究。分子 已知因素转录因子和指导提示等因素可以调节神经元的不同方面 开发，但是这些分子因子的作用尚未在中间神经元中进行深入研究 树突开发，突触伴侣的选择和电路形成。我将测试中心假设 时间转录因子调节引导受体表达，这建立了特定的树突状 形态和突触连接。为了研究果蝇神经索中的这一中心假设 将使用神经细胞3-3干细胞和它作为模型生成的EL中间神经元。在AIM 1中，我将测试 NB3-3，时间转录因子，CA和NAB中的工作假设激活不同的指导集 提示受体在其早期出生和后期的EL女儿中间神经元中，这决定了差异 树突定位。我使用单细胞RNA测序，NB3-3的CA和NAB的特异性基因操纵，以及 共聚焦显微镜检查树突形态。我希望连接时间转录的角色 引导提示受体表达的作用的因素。此外，我希望引入创新 测序方法我们当前对体感电路发展的分析。在AIM 2中，我将测试 工作假设是，在早期出生的El dendrites中必须使用指导受体roundabout3（robo3） 并且足以在后期的El树突中调节树突形态发生和位置。我会表演robo3 亚细胞定位分析以及ROBO3的获得和功能丧失测定法检查的功能 El Dendrites中的Robo3。此目标将有助于确定中间神经元中ROBO3的要求并开发新的 分析ROBO3本地化的测定。总体而言，该建议将连接到以前的不同领域 体感电路的发展。这项工作还将使未来的研究能够调查广泛的研究 神经元电路规范的保护和神经发育病理疗法的发展 其中电路接线被破坏。