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 正确发展的体感回路对于任何动物感知并与其互动都至关重要 环境。体感回路的发展要求回路内的每个神经元正确地 定位其轴突和树突。这使得它们能够建立特定的突触连接，并使电路能够 集合。在体感回路中，中间神经元是数量最多的神经元类型。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，中间神经元 由于缺乏有关体感电路架构的工具和知识，目前正在研究中。分子 已知诸如时间转录因子和引导信号之类的因子可以调节神经元的不同方面 然而，这些分子因素在中间神经元中的作用尚未得到深入研究 树突发育、突触伙伴选择和电路形成。我将检验中心假设 时间转录因子调节引导受体的表达，从而建立特定的树突 形态和突触连接。为了研究果蝇神经索的这一中心假设，我 将使用神经母细胞 3-3 干细胞及其生成的 EL 中间神经元作为模型。在目标 1 中，我将测试 NB3-3 中的工作假设、时间转录因子 Cas 和 Nab 激活不同的指导集 其早生和晚生 EL 子代中间神经元中的提示受体，这决定了差异 树突定位。我使用单细胞 RNA 测序、Cas 和 Nab 的 NB3-3 特异性基因操作，以及 共焦显微镜检查树突形态。我希望将时间转录的作用联系起来 指导线索受体表达作用的因素。此外，我希望引入一种创新的 我们目前对体感电路开发的分析采用测序方法。在目标 2 中，我将测试 工作假设：引导受体 Roundabout3 (Robo3) 在早期出生的 EL 树突中是必需的 并足以在晚生的 EL 树突中调节树突的形态发生和位置。我会表演Robo3 亚细胞定位测定以及 Robo3 的功能获得和丧失测定，以检查 EL 树突中的 Robo3。这一目标将有助于确定 Robo3 在中间神经元中的需求并开发新的 分析 Robo3 定位的分析方法。总体而言，该提案将与之前不同的领域相联系 体感电路开发。这项工作还将使未来的研究能够调查广泛的 神经元回路规范的保护和神经发育病理疗法的开发 其中电路布线被破坏。