INTERROGATING THE GENOME TO UNCOVER HUMAN SPECIALIZATIONS OF BRAIN & COGNITION

这个子项目是许多研究子项目中利用 资源由NIH/NCRR资助的中心拨款提供。子项目和 调查员(PI)可能从NIH的另一个来源获得了主要资金， 并因此可以在其他清晰的条目中表示。列出的机构是 该中心不一定是调查人员的机构。 人类大脑具有独特的功能能力，但人类大脑在结构和生化方面与其他灵长类动物的不同之处鲜为人知。我们正在通过识别不同物种(人类、黑猩猩和猕猴)、不同大脑区域(多个皮质和皮质下部位)以及不同半球之间的基因和蛋白质表达差异来解决这些问题。基因表达差异是使用短时间(8小时或更短)死后的未固定组织样本来确定的。样本被快速冷冻并储存在7080℃下。信使RNA随后被提取并与寡核苷酸微阵列杂交，以确定每个样本的基因表达水平。通过比较样本之间的微阵列结果来标记可能的物种差异和地区差异，然后使用定量实时RT-PCR来证实。蛋白质表达水平的差异由Western blotting确定。最后，通过原位杂交和免疫细胞化学方法对大脑皮质组织中的mRNA和蛋白质进行定位。

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. We are pursuing 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 are flagged by comparing microarray results across samples, and then are corroborated using quantitative real-time RT-PCR. Differences in levels of protein expression are determined by Western blotting. Finally, mRNA and protein are localized in cortical tissue using in situ hybridization and immunocytochemistry.