Mapping Neural Connectivity in Zebrafish Larvae Using a Photoconvertible Protein

项目总结 全脑功能连接的建立将促进对记忆的整体理解 形成，这是现代神经科学的一个主要目标。从了解职能部门获得的信息 神经元之间的连接和连接之间的潜在可塑性可以用来理解 治疗神经和精神疾病。 利用现有技术进行活体全脑成像需要具有优势的生物学特性 与强大的分子工具箱相结合。两种无脊椎动物模式生物具有这些特性：D. 黑腹蛇和线虫。然而，斑马鱼(Danio Rerio)是唯一一种有这种症状的脊椎动物 容量。与斑马鱼幼体形成功能连接体的能力提供了明显的优势 在人类疾病的动物模型中了解记忆的形成或精确定位异常神经回路。 此外，斑马鱼的快速发展、体型小、繁殖力高，使其成为高度养殖的理想生物。 通过筛查，发现治疗这些疾病的新疗法的有用机制。 该项目将尝试创造一种新的工具，在体内自由地快速形成全脑连接 移动鱼的长期目标是在电路水平上了解大脑功能和功能障碍。朝向 这个目标，我们将创造转基因鱼，在活性控制下表达一种可光转换的蛋白质-- 从属推动者。然后我们将确定每一条转基因鱼的实验程序 最有效和准确地反映正在调查的神经活动，并将与以下方面相关的信号降至最低 非特异性神经活动。在我们定义了我们的转基因的实验潜力和限制之后 我们将寻求确定记录的神经活动是否在生理上有效。要做到这一点，我们将 根据预先定义的感觉系统刺激，形成神经活动的功能图 神经连接。这些实验应该提供开始调查变化所需的试点数据 在斑马鱼神经发育障碍模型中由于记忆形成而产生的功能连接。 来自拟议研究的数据有望促进开发有效的治疗方法 大脑疾病和障碍，包括阿尔茨海默病、创伤后应激障碍、精神分裂症和自闭症。

PROJECT SUMMARY Creation of whole-brain functional connectomes will facilitate a holistic understanding of memory formation, which is a major goal of modern neuroscience. Information gained from understanding the functional connections between neurons and the underlying plasticity between connections can be used to understand and treat neurological and mental disorders. In vivo whole-brain mapping with current technologies requires advantageous biological attributes in combination with a substantial molecular toolbox. Two invertebrate model organisms have these properties: D. melanogaster and C. elegans. However, zebrafish (Danio rerio) is the only vertebrate species with this capacity. The ability to form a functional connectome with larval zebrafish affords distinct advantages to understanding memory formation or pinpointing aberrant neural circuits in animal models of human disorders. Further, rapid development, small size, and high fecundity of zebrafish makes them an ideal organism for high- throughput screening, a useful mechanism to discover novel therapeutics for these disorders. The project will attempt to create a new tool to rapidly form in vivo whole-brain connectomes in freely moving fish with the long-term goal of understanding brain function and dysfunction at a circuit-level. Towards this goal, we will create transgenic fish that express a photoconvertible protein under the control of activity- dependent promoters. We will then determine the experimental procedures for each line of transgenic fish that most efficiently and accurately reflect the neural activity under investigation and minimize signals related to nonspecific neural activity. After we have defined the experimental potential and constraints of our transgenic lines of fish, we will seek to determine if the recorded neural activity is physiologically valid. To do so, we will form functional maps of neural activity in response to stimulation of sensory systems with previously defined neural connections. These experiments should provide the pilot data necessary to begin investigating changes in functional connectomes due to memory formation in zebrafish models of neurodevelopmental disorders. Data from the proposed studies are expected to facilitate the development of effective treatments for brain diseases and disorders, including Alzheimer's disease, PTSD, schizophrenia and autism.