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PAR-22-093，我们提出了本项目，以解决几个紧迫的需求 在外地跨越不同年龄和性别的小鼠模型通常用于定量解剖学、分子学、生物学和分子生物学， 和神经退行性疾病如阿尔茨海默病（AD）的病理变化。然而，唯一可用的 标准小鼠脑图谱由2个月大的成年雄性小鼠构建。此外，虽然两者 正常衰老和AD神经变性显示性二态性特征，科学界缺乏 性分化啮齿动物大脑图谱研究这些属性所必需的。通过应用尖端技术 技术，我们已经开发了脑倡议连接和细胞类型映射项目，我们将（1） 用颗粒海马（HPF）分子生成性二态3D衰老和AD脑图谱 域，可用作所有HPF工作的标准图谱模板，以及（2）我们将全面 表征衰老和AD中的形态学营养不良以及连接和突触破坏。在 具体目标1，将创建老化和AD大脑的公开可用的标准3D HPF图谱。数据将 在2、9和18月龄野生型（WT）、5xFAD（早期AD发作）和MAPT（H1）* N279 K（晚期AD）中产生 发病）雄性和雌性小鼠。细HPF域描绘将通过细胞的3D体积图像来促进， 和骨髓结构，而其他组织病理学标志物（Aβ斑块/tau缠结）和 将绘制化学结构细节（谷氨酸、GABA、PV、SST、Calb 1），以创建全面的 组织病理学和化学结构HPF图谱。在具体目标2中，我们将系统地应用遗传学 MORF 3稀疏标记方法标记，重建和分析所有细胞类型特异性神经元形态 在WT、VAP1.M0RF3/5xFAD、PV.M0RF3/5xFAD中，HPF区域在其结构域的颗粒水平上， M0 RF 3/MAPT（H1）* N279 K和PV.M0RF3/MAPT（H1）* N279 K小鼠。我们整个 将应用脑3D清除、免疫染色、成像和3D神经元重建流水线。鉴于 AD相关认知功能减退的病因是选择性HPF突触破坏， 在具体目标3中，一些最早受AD影响的人将检查沿着的进行性连接中断。 淀粉样蛋白和tau病理学进展。雄性和雌性MORF 3和双转基因小鼠，MORF 3/5xFAD 和MORF 3/MAPT（H1）* N279 K，在2 m，9 m和18 m年龄，将用于揭示潜在的连通性变化 和AD之间的关系相同的组将用于确定突触水平的HPF破坏， 扩展显微镜的应用，将捕捉突触连接的超分辨率图像。在 具体目标4，我们将创建一个基于Web的数据门户，使可视化，比较和分析， 3D老化和AD大脑中的神经回路和细胞类型。我们的团队在连接组学方面有着数十年的经验， 大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，并确定性二态性解剖区域在年龄和AD进展中的脆弱性。

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.