Meso-Seq for in-depth transcriptomics in ultra-low amounts of FACS-purified neuronal nuclei. Meso-Seq for in-depth transcriptomics in ultra-low amounts of FACS-purified neuronal nuclei

Profiling of gene expression in sparse populations of genetically defined neurons is essential for dissecting the molecular mechanisms that control the development and plasticity of neural circuits. However, current transcriptomic approaches are ill suited for detailed mechanistic studies in sparse neuronal populations, as they either are technically complex and relatively expensive (e.g., single-cell RNA sequencing [RNA-seq]) or require large amounts of input material (e.g., traditional bulk RNA-seq). Overall design: Visual cortex mRNA profiles of adult mice' nuclear RNA, including (1) RNA-Seq of neurons sparsely labeled with AAV-constructs, (2) RNA-Seq from low amounts of antibody-labeled neuronal nuclei in the cortex of mice and of non-human primates, and (3), analyses of experience-regulated gene programs in sparse visual cortex neurons.

对经遗传标记定义的稀疏神经元群体进行基因表达谱分析，对于解析调控神经环路发育与可塑性的分子机制至关重要。然而，当前的转录组学方法并不适用于稀疏神经元群体的精细机制研究，因为这些方法要么技术复杂度高且成本相对高昂（例如单细胞RNA测序（RNA-seq）），要么需要大量起始实验材料（例如传统批量RNA测序）。实验整体设计：成年小鼠视觉皮层细胞核RNA的mRNA表达谱分析，涵盖以下三部分：(1) 经腺相关病毒（AAV）载体稀疏标记的神经元的RNA测序；(2) 小鼠及非人灵长类动物皮层中少量抗体标记神经元细胞核的RNA测序；(3) 稀疏标记视觉皮层神经元的经验依赖型基因表达程序分析。