INTERROGATING THE GENOME TO UNCOVER HUMAN SPECIALIZATIONS OF BRAIN & COGNITION

这个子项目是许多利用 由NIH/NCRR资助的中心赠款提供的资源。子项目和 研究者（PI）可能从另一个NIH来源获得了主要资金， 因此可在其他CRISP条目中表示。所列机构为 研究中心，而研究中心不一定是研究者所在的机构。 人类大脑具有独特的功能，但人类大脑在结构和生物化学上与其他灵长类动物的大脑有何不同，人们对此知之甚少。 在本报告所述期间，我们通过识别物种（人类、黑猩猩和猕猴）之间、大脑区域（多个皮质和皮质下部位）之间以及半球之间的基因和蛋白质表达差异来研究这些问题。 基因表达差异使用来自短死后时间（8小时或更少）病例的未固定组织样本来确定。 将样品快速冷冻并储存在 7080摄氏度 随后提取信使RNA并与寡核苷酸微阵列杂交以确定每个样品的基因表达水平。 可能的物种差异和区域差异通过比较样品中的微阵列结果来标记，然后使用定量实时RT-PCR来证实。 蛋白质表达水平的差异通过蛋白质印迹法测定。 最后，mRNA和蛋白质定位在皮质组织中使用原位杂交和免疫细胞化学。

This subproject is one of many research subprojects utilizing the resources provided by a Center grant funded by NIH/NCRR. The subproject and investigator (PI) may have received primary funding from another NIH source, and thus could be represented in other CRISP entries. The institution listed is for the Center, which is not necessarily the institution for the investigator. Human brains have unique functional capacities, but how the human brain differs structurally and biochemically from that of other primates is poorly understood. During the reporting period, we pursued these questions by identifying differences in gene and protein expression between species (humans, chimpanzees, and macaque monkeys), between brain regions (multiple cortical and subcortical sites), and between hemispheres. Gene-expression differences are determined using unfixed tissue samples from short post-mortem time (8 hours or less) cases. Samples are flash-frozen and stored at 7080¿C. Messenger RNA is later extracted and hybridized to oligonucleotide microarrays to determining gene-expression levels for each sample. Possible species differences and regional differences were flagged by comparing microarray results across samples, and then are corroborated using quantitative real-time RT-PCR. Differences in levels of protein expression were determined by Western blotting. Finally, mRNA and protein were localized in cortical tissue using in situ hybridization and immunocytochemistry.