Odor Memory Traces in the Mouse Olfactory Cortex - Supplement

项目摘要 学习和记忆是大脑的基本功能，但它们的基本细胞和神经回路 机制仍然知之甚少。气味记忆在人类和动物中异常强大， 对生存和繁殖具有突出的重要性，并且极易患上神经退行性疾病 包括阿尔茨海默病。长期以来，人们一直认为嗅觉(梨状)皮质编码气味记忆， 然而，嗅觉学习的细胞底物和电路机制尚不清楚。我们的长期目标 是为了了解气味感知和记忆的细胞和神经回路机制。这样做的目的是 提议是提供一种机械的细胞/分子理解气味记忆是如何编码的 表达。我们已经在老鼠身上开发了基于活动的交叉遗传方法，使我们能够识别 并操纵在嗅觉学习过程中被激活的梨状神经元的活动。目标1：确定 操纵气味记忆痕迹细胞的活动如何改变行为。我们将使用基于以下因素的基因标签 CFOS启动子活性(“Fos标记”)，以可视化和操纵梨状神经元的活性 在嗅觉学习过程中被激活。目标2：确定嗅觉学习如何改变气味反应 梨形合奏的性质。我们将在清醒状态下对气味诱发的活动进行慢性双光子成像， 在厌恶和嗜好的嗅觉条件作用之前、期间和之后的小鼠行为。我们将测试 嗅觉学习选择性地增强刺激检测和辨别能力编码的假说 构成嗅觉记忆痕迹的神经元。目标3：确定分子同一性和连接性 嗅觉记忆痕迹细胞。使用我们之前确定的一组标记基因和单细胞转录 我们将确定在学习过程中被激活的梨状神经元的分子身份，我们将追踪 他们的轴突投射。我们提出了一种多层次的实验方法来确定细胞底物 嗅觉学习和记忆，提供机械洞察神经电路功能是如何塑造的 经验。 要实现我们的目标，需要生成和分析大型多模式数据集，包括体内慢性数据集 钙成像和行为监测。我们将开发强大的软件工具，用于数据集成、分析、 存储和共享。通过与软件工程和计算神经科学专家的合作，我们 为数据处理构建了开放源码的标准化代码和文件格式。我们将增强代码的健壮性 和集成，我们将建立云兼容性。我们将调整我们的管道以适应一系列 开源软件工具，并提供图形用户界面。我们提出的创新软件解决方案 将提高整合的神经活动和行为数据的重复性和共享性，具有重要意义 一个庞大的神经科学用户群体。

Project Summary Learning and memory are fundamental brain functions, yet their underlying cellular and neural circuit mechanisms remain poorly understood. Odor memories are exceptionally robust in humans and animals, of outstanding importance for survival and reproduction, and highly susceptible to neurodegenerative disorders including Alzheimer Disease. The olfactory (piriform) cortex has long been suggested to encode odor memories, however, the cellular substrates and circuit mechanisms of olfactory learning are unknown. Our long-term goal is to understand the cellular and neural circuit mechanisms of odor perception and memory. The objective of this proposal is to provide a mechanistic cellular/molecular understanding of how odor memories are encoded and expressed. We have developed activity-based intersectional genetic approaches in mice that allow us to identify and manipulate the activity of piriform neurons that were activated during olfactory learning. Aim 1: To determine how manipulating the activity of odor memory trace cells alters behavior. We will use genetic tagging based on cFos promoter activity (“Fos-tagging”) to visualize and manipulate the activity of piriform neurons that were activated during olfactory learning. Aim 2: To determine how olfactory learning alters the odor response properties of piriform ensembles. We will perform chronic two-photon imaging of odor-evoked activity in awake, behaving mice, before, during, and after aversive and appetitive olfactory conditioning. We will test the hypothesis that olfactory learning selectively enhances the encoding of stimulus detection and discriminability in neurons constituting an olfactory memory trace. Aim 3: To determine the molecular identity and connectivity of olfactory memory trace cells. Using our previously identified set of marker genes and single cell transcriptomics we will determine the molecular identities of piriform neurons that are activated during learning, and we will trace their axonal projections. We propose a multi-tiered experimental approach to identify the cellular substrates for olfactory learning and memory, to provide mechanistic insight into how neural circuit functions are shaped by experience. Achieving our goals requires the generation and analysis of large, multimodal data sets, including chronic in vivo calcium imaging and behavioral monitoring. We will develop robust software tools for data integration, analysis, storage, and sharing. In collaboration with experts in software engineering and computational neuroscience we have built open-source standardized code and file formats for data processing. We will enhance code robustness and integration, and we will establish cloud compatibility. We will adapt our pipeline for usage with a range of open-source software tools and provide a graphical user interface. The innovative software solutions we propose will enhance reproducibility and shareability of integrated neural activity and behavioral data, with significance to a large neuroscience user group.