Bernal J, Godbout M, Hasel K W, Travis G H, Sutcliffe J G
Department of Molecular Biology, Research Institute of Scripps Clinic, La Jolla, California.
J Neurosci Res. 1990 Oct;27(2):153-8. doi: 10.1002/jnr.490270205.
A pool of 163 clones, isolated by screening 60,000 members of a Macaca fascicularis cerebral cortex cDNA library with a cortex-minus-cerebellum subtracted probe prepared by the phenol enhancement method, was analyzed by Northern blot hybridization studies. One hundred fifty-three of these clones corresponded to 22 RNAs whose abundance was at least 2-fold higher in cerebral cortex poly(A)+ RNA samples than in samples of cerebellar poly(A)+ RNA. Seven of these RNAs, represented by 131 clones, were undetectable in cerebellum. Only 10 of the 163 clones proved to be false positives. The abundance of several of these cortex-enriched RNAs was altered in Alzheimer's disease brains. Several RNAs that were present in cerebral cortex but undetectable in cerebellum were generally enriched in telencephalon, although none was restricted to the cortex. One of the cortex enriched RNAs, whose nucleotide sequence is presented, encoded monkey preprocholecystokinin. Overall, this study provides insights into the powers and limitations of subtractive hybridization and into the patterns of gene expression in the central nervous system.
通过用酚增强法制备的皮质减小脑扣除探针筛选60000个猕猴大脑皮质cDNA文库成员，分离出163个克隆。通过Northern印迹杂交研究对其进行了分析。其中153个克隆对应于22种RNA，其在大脑皮质聚腺苷酸加（poly(A)+）RNA样品中的丰度至少比小脑聚腺苷酸加（poly(A)+）RNA样品中的丰度高2倍。其中7种RNA（由131个克隆代表）在小脑中无法检测到。163个克隆中只有10个被证明是假阳性。这些大脑皮质富集的RNA中有几种在阿尔茨海默病大脑中的丰度发生了改变。几种存在于大脑皮质但在小脑中无法检测到的RNA通常在端脑中富集，尽管没有一种仅限于皮质。其中一种大脑皮质富集的RNA（其核苷酸序列已给出）编码猴前胆囊收缩素原。总体而言，这项研究深入了解了扣除杂交的能力和局限性以及中枢神经系统中的基因表达模式。
