A New Animal Model to Examine Nervous System Function Development and Regeneration

模式生物已被证明是非常宝贵的，因为它是研究生物学基本原理的更易处理的系统，并通过研究进化创新而取得突破，例如水母的GFP或细菌的CRISPR。我的目标是利用一个新的模型系统，在我的博士后工作中开发的，这是独特的定位，以提供这两种类型的见解：Clytia hemisphaerica，一种地中海水母。 该提案使用Clytia的独特特征来确定神经发育，神经再生和系统神经科学界面的精确机制。首先，Clytia很小（<1 mm-1 cm），透明，遗传上易于处理，使得使用遗传编码光学技术在体内同时成像和操纵神经系统中每个神经元的活动成为可能。此外，Clytia在其生命周期内将其神经系统的数量级扩展至少一个数量级，而不会破坏系统功能，最终超过一万个神经元。因此，还可以在整个生物体中同时观察连续分化、迁移、轴突靶向和功能活性。最后，Clytia具有鲜为人知的强大再生能力。这些包括大量基因消融神经元的再生，以及这些神经元控制的行为的快速恢复。·系统神经科学中的基本问题，包括行为和行为状态的机制，神经调节的作用，以及研究系统组织和跨尺度功能的方法。 神经发育的基本原则，特别是在发育和功能的界面。 ·使再生和新神经元无缝整合到功能网络中的机制。在目标1中，我将描述神经元的分子表型，并建立基于CRISPR的基因敲入，将效应子靶向特定的细胞亚群。在目标2中，我将使用协调行为作为切入点，开发和测试潜在神经机制的模型。在目标3中，我将研究这种行为系统如何对持续的神经发生保持稳健，以及在神经元基因消融后使其快速恢复的机制。 我的愿景是，一旦关键的基础工作完成并发表，Clytia将成为一个广泛使用的模型系统。这个提议是第一步，为我未来的独立项目和更广泛的Clytia社区奠定了基础，并提供了必要的培训，我需要填补我的知识空白，专注于计算和单细胞RNA测序方法。

Model organisms have proven invaluable as more tractable systems to study fundamental principles of biology and by yielding breakthroughs that result from studying evolutionary innovations, for example, GFP, from jellyfish, or CRISPR, from bacteria. My goal is to leverage a new model system, developed during my postdoctoral work, that is uniquely positioned to provide insights of both types: Clytia hemisphaerica, a species of Mediterranean jellyfish. This proposal uses Clytia's distinctive features to identify precise mechanisms at the interface of neural development, neural regeneration, and systems neuroscience. First, Clytia are tiny (<1mm-1cm), transparent, and genetically tractable, making it possible to image and manipulate the activity of every neuron in the nervous system simultaneously, in vivo, using genetically encoded optical techniques. Further, Clytia numerically scale their nervous system at least an order of magnitude during their lifespan without disrupting system function, ending with more than ten thousand neurons. It is therefore also possible to observe continuous differentiation, migration, axon targeting, and functional activity simultaneously across the whole organism. Lastly, Clytia have poorly understood and powerful regenerative capabilities. These include the regeneration of large populations of genetically ablated neurons, with rapid recovery of the behaviors that those neurons control. These properties make Clytia an experimentally tractable platform to investigate: · Fundamental questions in systems neuroscience, including mechanisms underlying behaviors and behavior states, roles of neuromodulation, and approaches for studying system organization and function across scales. · Basic principles of neurodevelopment, particularly at the interface of development and function. · Mechanisms enabling regeneration and the seamless integration of new neurons into a functioning network. In Aim 1, I will characterize the molecular phenotypes of neurons and establish CRISPR-based knock-in to target effectors to specific subpopulations of cells. In Aim 2, I will use a coordinated behavior as my point of entry and develop and test models of underlying neural mechanisms. In Aim 3, I will examine how this behavioral system is robust to constant neurogenesis, and the mechanisms that enable its rapid recovery following genetic ablation of neurons. My vision is that, once the key foundational work has been completed and published, Clytia will become a widely used model system. This proposal serves as the first step, laying the foundation for my future independent program and for a Clytia community more broadly, and providing the essential training that I need to fill gaps in my knowledge, focusing on computational and single-cell RNA sequencing approaches.