Sexual dimorphic cell type and connectivity atlases of the aging and AD mouse brains

项目摘要 为了响应NOT-21-039和相关的PAR-22-093，我们提出这个项目来解决几个紧急需求 在战场上。跨越不同年龄和性别的小鼠模型通常被用来量化解剖，分子， 以及阿尔茨海默病(AD)等神经退行性疾病的病理变化。然而，唯一可用的 标准的小鼠脑图谱是从2个月大的成年雄性小鼠身上构建的。此外，虽然两者 正常衰老和阿尔茨海默病神经退行性变表现出性二态特征，科学界缺乏 研究这些属性所需的性别分化啮齿动物脑图谱。通过应用尖端技术 我们为大脑倡议连接和细胞类型映射项目开发的技术，我们将(1) 用颗粒海马区(HPF)分子生成性二态3D衰老和AD脑图谱 域名，可用作所有HPF工作的标准图集模板和(2)我们将全面 描述衰老和阿尔茨海默病的形态障碍，以及连接和突触中断。在……里面 具体目标1，将创建公开可用的标准3D HPF老化和AD大脑地图集。数据将是 在2个月、9个月和18个月大的野生型(WT)、5xFAD(早期AD)和MAPT(H1)*N279K(晚期AD)中生成 发病)雄鼠和雌鼠。精细的HPF结构域的勾画将通过细胞的3D体积图像来实现。 和骨髓结构，而其他组织病理学标记(Aβ斑块/tau缠结)和 化学结构细节(谷氨酸、GABA、PV、SST、Calb1)将被映射以创建全面的 组织病理学和化学构筑的HPF图谱。在具体目标2中，我们将系统地应用一种基因 MORF3稀疏标记方法标记、重建和分析ALL细胞类型特定的神经元形态 在WT、Vgu1.MORF3/5xFAD、PV.MORF3/5xFAD、 不同年龄和性别的小鼠，分别为：1.MORF3/MAPT(H1)*N279K和PV.MORF3/MAPT(H1)*N279K。我们的整个 将应用脑3D清除、免疫染色、成像和3D神经元重建管道。考虑到 AD相关认知功能下降的原因之一是选择性HPF突触中断，HPF背侧结节是 在特定目标3中，一些最早受到AD影响的患者将检查沿途的渐进性连接中断 淀粉样蛋白和tau病理进展。雄性和雌性MORF3和双转基因小鼠，MORF3/5xFAD 和MORF3/MAPT(H1)*N279K，年龄分别为2m、9m和18m，将用于揭示潜在的连通性变化 跨越老龄化和AD。相同的组将用于确定突触水平的HPF中断 应用扩展显微镜捕捉突触连接的超分辨率图像。在……里面 具体目标4，我们将创建一个基于Web的数据门户，以实现可视化、比较和分析 3D老化和AD脑中的神经回路和细胞类型。我们的团队，在连接学方面有几十年的经验， 脑图谱和在线可视化肯定会为所有神经科学提供标准的HPF图谱模板 研究并确定易受年龄和AD进展影响的性二型解剖区域。

Project Abstract In response to NOT-21-039 and related PAR-22-093, we propose this project to address several urgent needs in the field. Mouse models spanning different ages and sexes are routinely used to quantify anatomic, molecular, and pathologic changes in neurodegenerative diseases like Alzheimer’s Disease (AD). Yet, the only available standard mouse brain atlases are constructed from 2-month-old adult male mice. Furthermore, although both normal aging and AD neurodegeneration display sexually dimorphic features, the scientific community lacks the sexually differentiated rodent brain atlases necessary to study these attributes. By applying cutting edge technologies, we have developed for BRAIN Initiative connectomic and cell type mapping projects, we will (1) generate sexually dimorphic 3D aging and AD brain atlases with granular hippocampus (HPF) molecular domains, that can be used as standard atlas templates for all HPF work and (2) we will comprehensively characterize morphological dystrophies, as well as connectional and synaptic disruptions, in aging and AD. In Specific Aim 1, publicly available standard 3D HPF atlases of aging and AD brains will be created. Data will be generated in 2-, 9-, and 18-month-old wildtype (WT), 5xFAD (early AD onset), and MAPT(H1)*N279K (late AD onset) male and female mice. Fine HPF domain delineations will be facilitated by 3D volumetric images of cyto- and myeloarchitecture, while additional histopathological markers (Aβ plaques/tau tangles) and chemoarchitectural details (glutamate, GABA, PV, SST, Calb1) will be mapped to create comprehensive histopathological and chemoarchitectural HPF atlases. In Specific Aim 2, We will systematically apply a genetic MORF3 sparse labeling approach to label, reconstruct, and analyze cell type specific neuronal morphology of all HPF regions at the granular level of their domains in WT, Vglut1.MORF3/5xFAD, PV.MORF3/5xFAD, Vglut1.MORF3/MAPT(H1)*N279K, and PV.MORF3/MAPT(H1)*N279K mice across age and sex. Our whole brain 3D clearing, immunostaining, imaging, and 3D neuronal reconstruction pipeline will be applied. Given that an etiology of AD-related cognitive decline is selective HPF synaptic disruptions, with dorsal HPF nodes being some of the earliest affected in AD, in Specific Aim 3, will examine progressive connectional disruptions along amyloid and tau pathology progression. Male and female MORF3 and double transgenic mice, MORF3/5xFAD and MORF3/MAPT(H1)*N279K, at 2m, 9m, and 18m of age will be used to reveal potential connectivity changes across aging and AD. The same groups will be used to determine synaptic-level HPF disruptions with the application of Expansion Microscopy that will capture super-resolution images of synaptic connections. In Specific Aim 4, we will create a web-based data portal that enables visualization, comparison, and analysis of neural circuits and cell types in 3D aging and AD brains. Our team, with decades of experience in connectomics, brain atlasing, and online visualization, is sure to deliver standard HPF atlas templates for all neuroscience research and to determine sexually dimorphic anatomic regions vulnerable across age and AD progression.