A new animal model to examine nervous system function, development, and regeneration

事实证明，模式生物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产生的突破，例如来自水母的GFP，或来自细菌的CRISPR，被证明是非常宝贵的。我的目标是利用我在博士后工作期间开发的一个新的模型系统，该系统处于独特的位置，可以提供这两种类型的见解：Clytia hemisphaerica，一种地中海水母。这项建议利用Clytia的独特特征来识别神经发育、神经再生和系统神经科学界面上的精确机制。首先，Clytia很小(&lt；1 mm-1 cm)，透明，遗传上容易处理，使得使用遗传编码的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在它们的生命周期中，在不破坏系统功能的情况下，至少在数字上扩展它们的神经系统一个数量级，最终拥有超过一万个神经元。因此，也有可能同时观察到整个生物体的持续分化、迁移、轴突靶向和功能活动。最后，Clytia没有很好的理解和强大的再生能力。这些措施包括大量经过基因消融的神经元的再生，这些神经元控制的行为迅速恢复。这些特性使Clytia成为一个易于实验处理的平台，用于研究：·系统神经科学中的基本问题，包括行为和行为状态的潜在机制，神经调节的作用，以及跨尺度研究系统组织和功能的方法。·神经发育的基本原则，特别是在发育和功能的界面上。·能够再生和将新神经元无缝整合到一个正常运作的网络中的机制。在目标1中，我将描述神经元的分子表型，并建立基于CRISPR的针对特定细胞亚群的靶向效应。在目标2中，我将使用协调行为作为我的切入点，开发和测试潜在神经机制的模型。在目标3中，我将研究这一行为系统如何对持续的神经发生强大，以及使其能够在神经元遗传消融后迅速恢复的机制。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。这项提议是第一步，为我未来的独立计划和更广泛的Clytia社区奠定了基础，并提供了我需要的基本培训，以填补我的知识空白，重点是计算和单细胞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.