A three dimensional multimodal cellular connectivity atlas of the mouse hypothalamus

小鼠下丘脑三维多模态细胞连接图谱

• 批准号: 10719606
• 负责人: Hong-Wei Dong
• 金额: $ 65.22万
• 依托单位: UNIVERSITY OF CALIFORNIA LOS ANGELES
• 依托单位国家: 美国
• 财政年份: 2023
• 资助国家: 美国
• 起止时间: 2023-07-15 至 2028-05-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10719606
• 关键词: 3-Dimensional; Aggressive behavior; Algorithms; Anatomy; Androgen Receptor; Animals; Antibodies; Architecture; Arousal; Atlases; Axon; Behavior; Behavioral; Brain; Brain region; Catalogs; Circadian Rhythms; Classification; Data; Dendrites; Disease; Estrogen Receptors; Feeding behaviors; Female; Foundations; Functional disorder; Genes; Genetic; Glutamates; Goals; Heterogeneity; Histologic; Homeostasis; Human; Hypothalamic structure; Immunohistochemistry; Individual; Informatics; Internal Ribosome Entry Site; Investigation; Knock-in Mouse; Label; Lateral Hypothalamic Area; Light; Manuals; Maps; Metabolism; Methods; Microscope; Microscopy; Modeling; Molecular; Monsters; Morphology; Mus; Neural Pathways; Neurons; Neuropeptides; Organizational Productivity; Pathway interactions; Physiologic Thermoregulation; Play; Process; Property; Publishing; Rabies; Reporter; Reproductive Behavior; Resolution; Role; Social Behavior; Specificity; Structure; Synapses; Taxonomy; Techniques; Testing; Tracer; Undifferentiated; Viral; Visualization; calbindin; cell type; cloud based; computational platform; computer infrastructure; data exchange; data portal; design; excitatory neuron; hypocretin; inhibitory neuron; innovation; male; microscopic imaging; multimodality; network architecture; neural; novel; reconstruction; reproductive; sex; sexual dimorphism

Project Summary/Abstract The mammalian hypothalamus plays a critical role in behaviors essential for survival. The lateral hypothalamic area (LHA) occupies 55% of the hypothalamus and contains diverse neuron types, yet remains an undifferentiated region in mouse atlases. Much remains unknown about its structural and functional organization at the regional, cell-type specific, and individual neuron scales. Further, the hypothalamus is structurally and functionally sexually dimorphic, yet differentiated brain atlases for the sexes is absent. We hypothesize that multiscale heterogeneity of LHA structural organization underlies the specificity and properties of its cell type- specific circuit functional organization. We will investigate this using established and innovative approaches to correlate and synthesize molecular, cellular, and circuit-level LHA data into a multimodal, multiscale LHA reference atlas. In Aim 1, we will construct a multiscale, multimodal, 3D reference atlas of the male and female mouse hypothalamus based on the cyto- and chemoarchitecture of the LHA (Aim 1a). These volumetric 3D lightsheet data will be registered to the Allen CCF and structural delineations will be conducted using a novel cloud-based 3D approach. In Aim 1b, we will use our connectivity-based parcellation strategy to refine the delineation of the LHA at a much higher granularity based on the systematic analysis of >300 sets of pre-collected neural pathways and additional connectivity data collected via conditional (Cre-dependent) viral pathway tracing methods. The refined parcellation of LHA subdivisions will be validated by investigating specificities of their input/output organization. In Aim 2, we will systematically investigate LHA molecular diversity and network organization across meso- and microscale resolutions. First, for each LHA subdivision, we will investigate the molecular identities of target-specific GABA and GLU projection neurons by combining retrograde pathway labeling with RNAscope or HiPlex RNAscope (Aim 2a). Then, for these target-specific GABA & GLU neuron types, we will systematically examine their axon collateralizations (Aim 2b) and neural inputs (Aim 2c) using a 2- way viral labeling method in Vgat-Cre and Vglut2-Cre knock-in mice. Finally, in Aim 2d, by combining a novel genetic sparse labeling method (MORF) with brain clearing, 3D microscopic imaging, and computational reconstruction algorithms, we will reconstruct the fine detailed single neuron morphology (dendrites and axons) of >2000 GABA and GLU neurons in male and female LHA. All of the data will be registered into the newly constructed 3D hypothalamic atlas (Aim 1) within the CCF to construct an unprecedentedly comprehensive cellular atlas of the male and female hypothalamus. In Aim 3, we will establish a computational platform and a specialized computational infrastructure for (1) circuit, neuronal, and synaptic reconstruction, annotation, and error correction validated with efficient ground truth human assessments; and (2) for efficient data transfer, storage, and dissemination. All CCF-registered neuronal morphology and connectivity data will be processed and disseminated through our UCLA BRAIN data portal.

项目摘要/摘要 哺乳动物下丘脑在生存至关重要的行为中起关键作用。下丘脑侧 面积（LHA）占下丘脑的55％，包含多种神经元类型，但仍然是 小鼠地图酶中未分化的区域。关于其结构和功能组织仍未清楚 在区域，细胞型特异性和单个神经元尺度上。此外，下丘脑在结构上是 在功能上具有性二态性，但不存在分化性别的大脑图谱。我们假设这一点 LHA结构组织的多尺度异质性是其细胞类型的特异性和特性 - 特定电路功能组织。我们将使用已建立和创新的方法进行调查 将分子，细胞和电路级LHA数据相关联和合成为多模式的LHA 参考地图集。在AIM 1中，我们将构建男女的多尺度，多模式，3D参考地图集 小鼠下丘脑基于LHA的细胞和化学结构（AIM 1A）。这些体积3D 灯表数据将注册到Allen CCF，结构描述将使用新颖 基于云的3D方法。在AIM 1B中，我们将使用基于连接性的划分策略来完善 基于对> 300组预制的系统分析，以更高的粒度描绘LHA 神经通路和通过条件（CRE依赖性）病毒途径收集的其他连接数据 方法。 LHA细分的精致分类将通过调查其特异性来验证 输入/输出组织。在AIM 2中，我们将系统地研究LHA分子多样性和网络 跨Meso和Microscale决议的组织。首先，对于每个LHA细分，我们将研究 通过结合逆行途径，目标特异性GABA和GLU投影神经元的分子身份 用rnascope或hiplex rnascope标记（AIM 2A）。然后，对于这些目标特异性GABA和GLU神经元 类型，我们将系统地检查它们的轴突抵押品（AIM 2B）和神经输入（AIM 2C）使用2- VGAT-CRE和VGLUT2-CRE敲入小鼠中的病毒标记方法。最后，在AIM 2D中，结合了一本小说 带有大脑清除，3D微观成像和计算的遗传稀疏标记方法（MORF） 重建算法，我们将重建精细的详细单神经元形态（树突和轴突） 男性和女性LHA中的> 2000 GABA和GLU神经元。所有数据将注册到新的 在CCF中建造的3D下丘脑图集（AIM 1），以建造一个前所未有的全面 雄性和女性下丘脑的细胞地图集。在AIM 3中，我们将建立一个计算平台和一个 （1）电路，神经元和突触重建，注释以及 通过有效的地面真相评估验证的错误纠正； （2）对于有效的数据传输， 存储和传播。将处理所有CCF注册的神经元形态和连接数据 并通过我们的UCLA脑数据门户传播。