A multimodal platform to bridge the experimental gap between behavioral, neuronal, and molecular studies

摘要 描述特定脑功能下的特定神经元身份和连接仍然是 神经科学的核心目标世纪以来，神经解剖学一直在 将神经元的突触接触、树突形态和轴突投射与其连接性联系起来。 遗传探针、光学成像模式和计算机技术的进步使得监测成为可能 并以前所未有的精度和规模操纵活体动物的神经活动。此外，本发明还提供了一种方法， “组学”研究的前沿开始区分异质性的分子多样性， 同一脑区的神经元独特分子标记的鉴定进一步 能够创建新的转基因模型来询问神经元的精确子集。尽管 将这些革命性的技术应用于研究系统和行为的巨大成功 神经科学，目前缺乏一个统一的实验范式，直接联系活动，连接 和分子信息的功能电路中完全相同的神经元在单细胞/单 突触分辨率这样做的能力将消除目前试图将不同的 从不同动物中取样的“相同”神经元群体的属性。更重要的是， 这样做将大大提高我们区分特定神经元的效率和准确性， 与同一大脑区域中不同回路功能相关的人群。 在这里，我们证明了coMAAP的可行性，这是一种多模态实验范式， 在相同的神经元中，相同神经元的活动、解剖和分子身份的相关光学映射， 动物重要的是，coMAAP可以使用标准仪器实现。同时结合 它可以实现前所未有的分辨率和规模。我们的目标 建议是优化和验证coMAAP实验范式，并利用coMAAP 描绘了在小鼠腹侧被盖中被唤醒激活的异源神经元群 区域（VTA）。

ABSTRACT Depicting the specific neuronal identity and connectivity underlying particular brain function remains a central goal for neuroscience. For over a century, neuroanatomy has continued to play critical roles in referencing a neuron's synaptic contact, dendritic morphology and axonal projection to its connectivity. The advances of genetic probes, optical imaging modalities and computer technologies permit monitoring and manipulating neuronal activity in living animals with unprecedented precision and scale. In addition, the forefront of “-omics” study begins to discriminate the molecular diversity of the heterogenous population neurons in the same brain region. The identification of unique molecular markers further enables creating novel transgenic models to interrogate precise subsets of neurons. Despite the tremendous success in applying these revolutionary technologies in studying systems and behavior neuroscience, there currently lacks a unified experimental paradigm to directly link activity, connection and molecular information of the exact same neurons in a functional circuit at the single cell/single synapse resolution. The ability to do so will remove the ambiguity in current attempts to correlate different attributes of the “same” neuronal populations sampled from different animals. More importantly, the ability to do so will tremendously improve our efficiency and accuracy in differentiating specific neuronal populations that correlate with distinct circuit functions in the same brain region. Here, we demonstrate the feasibility of coMAAP, a multimodal experimental paradigm that allows correlative optical mapping of activity, anatomy and molecular-identity of the same neurons in the same animal. Importantly, coMAAP can be implemented using standard instruments. While combining with specialized imaging modalities it can achieve unprecedented resolution and scale. The goals of our proposal are to optimize and validate the coMAAP experimental paradigm, and to utilize coMAAP to depict the heterogenous neuronal populations that are arousal activated in the mouse ventral tegmental area (VTA).