Profiling the human dentate gyrus across the lifespan with spatially-resolved transcriptomics

项目摘要 海马体在整个生命周期的学习和记忆中发挥着重要作用， 从婴儿期到老年观察到的发育和可塑性相关基因表达变化。由于 它的结构和电路，该地区已被链接到一些关键的行为功能。海马 根据树突-轴突极性和突触连接，神经元被组织成密集的层 海马体内是整个生命周期中结构和功能可塑性的主要场所， 与学习、记忆、情绪和压力调节相关的关键功能。许多重要的塑料相关 转录物定位于树突区室，并且它们的转录、运输出细胞核， 树突区室内的翻译受到发育和神经元活动的严格调节。 此外，尽管对出生后神经发生的功能重要性进行了广泛的表征， 啮齿动物齿状回（DG），成人DG（hDG）神经发生的无可争议的证据仍然存在 难以捉摸，其在整个生命周期中的持续性仍然存在争议。近年来，细胞类型特异性分子 已经描述了成年人和整个生命周期中的人类海马体的概况，使用单- 核RNA测序（snRNA-seq）;然而，这些资源缺乏空间分辨率， 海马和失去转录组信息从胞质区室。考虑到 HPC的空间结构和功能之间的关系，以及定位于HPC的转录本的特别重要性。 突触区室，分子分析技术，有能力解决这些差距，在我们的 知识是需要的。因此，我们建议产生空间分辨的，转录组范围的基因 在整个生命周期中hDG的表达谱，来自人类四个年龄组的神经型供体， 寿命（婴儿、青少年、中年和老年人）。数据将被编译成一个用户友好的浏览器， 社区资源。专家神经生物学家将执行规范子字段的手动注释， 生物统计学家将应用无监督聚类算法来确定新的空间域。然后我们将 比较整个生命周期的基因表达，以确定发育和空间调控的基因 表情我们将验证特定的新基因标记的细胞表达模式， smFISH在我们的供体队列中。这种方法将有助于空间基因表达模式的精细注释 在人类海马体中，有助于理解神经发育和神经退行性疾病 通过鉴定与空间上限定的细胞群的临床关联， 预防和治疗。

PROJECT SUMMARY The hippocampus plays a well-established role in learning and memory across the lifespan, with developmental and plasticity-related gene expression changes observed from infancy through old age. Due to its structure and circuitry, the region has been linked to a number of critical behavioral functions. Hippocampal neurons are organized in densely packed layers according to dendrite-axon polarity, and synaptic connections within the hippocampus are major sites of structural and functional plasticity across the lifespan that regulate critical functions related to learning, memory, mood and stress regulation. Many important plasticity-related transcripts are localized to the dendritic compartment, and their transcription, transport out of the nucleus, and translation within the dendritic compartment is tightly regulated, both developmentally and by neuronal activity. Additionally, despite extensive characterization of the functional importance of postnatal neurogenesis in rodent dentate gyrus (DG), indisputable evidence of adult neurogenesis in the human DG (hDG) remains elusive and its persistence throughout the lifespan remains controversial. Recently, cell-type specific molecular profiles of the human hippocampus in adults and across the lifespan have been described, using single- nucleus RNA sequencing (snRNA-seq); however, these resources lack spatial resolution within the hippocampus and lose transcriptomic information from the cytosolic compartment. Given the tight correlation between spatial structure and function in HPC, and the particular importance of transcripts localized to the synaptic compartment, molecular profiling technologies with the capacity to address these gaps in our knowledge are needed. Thus, we propose the generation of spatially-resolved, transcriptome-wide gene expression profiles in hDG across the lifespan, from neurotypical donors in four age groups across the human lifespan (infant, teen, middle-age, and elderly). Data will be compiled into a user-friendly browser as a community resource. Expert neurobiologists will perform manual annotations of canonical subfields, while biostatisticians will apply unsupervised clustering algorithms to identify novel spatial domains. We will then compare gene expression across the lifespan to identify developmentally- and spatially-regulated gene expression. We will validate the cellular expression patterns of specific novel gene markers by performing smFISH in our donor cohort. This approach will facilitate refined annotation of spatial gene expression patterns in the human hippocampus, and contribute to understanding neurodevelopmental and neurodegenerative disorders by identifying clinical associations with spatially-defined cell populations that can be targeted for prevention and treatment.