Metabolic adaptations in the brain are critical to the establishment and maintenance of normal cellular functions and to the pathological responses to disease processes. Here, we have focused on specific metabolic pathways that are involved in immune-mediated neuronal processes in brain using isolated neurons derived from human autopsy brain sections of normal individuals and individuals diagnosed as Alzheimer’s disease (AD). Laser capture microscopy was used to select specific cell types in immune-stained thin brain sections followed by NanoString technology to identify and quantify differences in mRNA levels between age-matched control and AD neuronal samples. Comparisons were also made between neurons isolated from AD brain sections expressing pathogenic hyperphosphorylated AT8- positive (AT8+) tau and non-AT8+ AD neurons using double labeling techniques. The mRNA expression data showed unique patterns of metabolic pathway expression between the subtypes of captured neurons that involved membrane based solute transporters, redox factors, and arginine and methionine metabolic pathways. We also identified the expression levels of a novel metabolic gene, Radical-S-Adenosyl Domain1 (RSAD1) and its corresponding protein, Rsad1, that impact methionine usage and radical based reactions. Immunohistochemistry was used to identify specific protein expression levels and their cellular location in NeuN+ and AT8+ neurons. APOE4 vs APOE3 genotype-specific and sex-specific gene expression differences in these metabolic pathways were also observed when comparing neurons from individuals with AD to age-matched individuals.

大脑中的代谢适应对于正常细胞功能的建立和维持以及对疾病过程的病理反应至关重要。在这里，我们使用来自正常个体和被诊断为阿尔茨海默病 (AD) 个体的人体尸检大脑切片的分离神经元，专注于参与大脑中免疫介导的神经元过程的特定代谢途径。激光捕获显微镜用于选择免疫染色的脑薄切片中的特定细胞类型，然后使用 NanoString 技术识别和量化年龄匹配的对照和 AD 神经元样本之间 mRNA 水平的差异。还使用双重标记技术对从表达致病性过度磷酸化 AT8 阳性 (AT8+) tau 和非 AT8+ AD 神经元的 AD 脑切片分离的神经元进行了比较。mRNA 表达数据显示了捕获的神经元亚型之间代谢途径表达的独特模式，这些亚型涉及基于膜的溶质转运蛋白、氧化还原因子以及精氨酸和蛋氨酸代谢途径。我们还鉴定了一种新的代谢基因 Radical-S-Adenosyl Domain1 的表达水平。RSAD1 ) 及其相应的蛋白质 Rsad1，它们会影响蛋氨酸的使用和基于自由基的反应。免疫组织化学用于鉴定特定蛋白质表达水平及其在 NeuN+ 和 AT8+ 神经元中的细胞位置。在比较 AD 个体与年龄匹配个体的神经元时，还观察到这些代谢途径中APOE4与APOE3基因型特异性和性别特异性基因表达的差异。