miRNA Control of Synaptic Stability and Structural Plasticity

项目摘要/摘要 让我着迷的是神经系统调节我们的适应性反应的能力。 不断变化的环境和我们不断变化的行为状态。作为进入这一现象的切入点，在我的 在研究生培训中，我使用了果蝇神经肌肉接头(NMJ)来建立应用专业知识 基因和分子分析，以了解神经回路如何适应生理下游 输入。在这种情况下，在细胞水平上，我最好奇的是神经元和它们的靶子如何适应 突触形态发生过程中高度专门化和互补性的细胞形态。在 在分子水平上，神经活动启动钙依赖转录和后转录的部署。 调节突触形态和组织的转录程序。在了解了以下因素后 控制突触间隔内的局部翻译，如脆性X智力低下蛋白 (FMRP)，我对SM的转录后控制非常感兴趣。从机械上讲，我对 在SM期间，单个基因的翻译是如何在神经活动的下游受到控制的，这让我 去研究miRNA。在我到目前为止的论文工作中，我已经确定了几个对活动至关重要的miRNAs- 包括miR-973在内的NMJ末端的依赖形态发生。MIR-973失去功能揭示了一种新的 表型，提示它是活性依赖的突触稳定性所必需的。 通过这些努力，我对描述变化的方法有了很好的理解 在突触结构和可塑性方面。在本建议书的F99阶段，我将通过使用 解决miR-973机制贡献的光遗传学、核糖体图谱和遗传学方法 在NMJ的突触稳定性方面。同时，我与我的赞助人和合作伙伴制定了培训计划- 赞助商，以确保我正在积极思考我的工作与突触可塑性的更广泛的相关性 哺乳动物的系统。总体而言，F99阶段的培训将提供坚实的基础，使 我希望建立有效的实验方法，在广泛的背景下研究神经可塑性。 展望未来，我将处于有利的地位，可以过渡到K00阶段，在那里我将在以下方面应用我的专业知识 成年哺乳动物脑中突触形态发生对运动介导的神经可塑性的影响。我的终极目标 是利用这种方法来识别新的分子通路和信号因子，这些分子通路和信号因子介导神经 可塑性作为治疗应用的潜在靶点，这与大脑倡议相一致 神经治疗学和塑造神经回路的机制。

PROJECT SUMMARY / ABSTRACT What fascinates me is the ability of the nervous system to mediate our adaptive responses to our changing environment and our changing behavioral states. As an entry point into this phenomenon, in my graduate training, I’ve used the Drosophila neuromuscular junction (NMJ) to build expertise in applying genetic and molecular analysis to understand how neural circuitry is adapted downstream of physiological input. In this context, at the cellular level, I’ve been most curious as to how neurons and their targets adapt highly specialized and complementary cellular morphologies during synaptic morphogenesis (SM). At the molecular level, neural activity initiates the deployment of calcium-dependent transcriptional and post- transcriptional programs that regulate synaptic morphology and organization. After learning about factors that control local translation within the synaptic compartment, such as the fragile X mental retardation protein (FMRP), I became very interested in post-transcriptional control of SM. Mechanistically, I am interested in how the translation of individual genes is controlled downstream of neural-activity during SM, which lead me to studying miRNAs. In my dissertation work thus far, I have identified several miRNAs as essential for activity- dependent morphogenesis of the NMJ terminal including miR-973. MiR-973 loss of function revealed a novel phenotype and suggested that it is required for activity-dependent synapse stability. Through these efforts, I have developed a sound understanding of approaches to characterize changes in synaptic architecture and plasticity. I will build upon this skill set in the F99 phase of this proposal by using optogenetics, ribosome profiling, and genetic approaches to resolve the mechanistic contribution of miR-973 in synaptic stability at the NMJ. Concurrently, I have developed a training plan with my sponsor and co- sponsor to ensure that I am actively thinking about the broader relevance of my work to synaptic plasticity in mammalian systems. Collectively, training during the F99 phase will provide a solid foundation that will enable me to construct effective experimental approaches to studying neural plasticity in a broad range of contexts. Moving forward, I will be in great position to transition into the K00 phase where I will apply my expertise in synaptic morphogenesis to exercise-mediated neural plasticity in the adult mammalian brain. My ultimate goal is to utilize this approach to identify novel molecular pathways and signaling factors that mediate neural plasticity as potential targets for therapeutic applications, which aligns with the Brain Initiative on Neurotherapeutics and Mechanisms shaping neuronal circuitry.