Travis G H, Naus C G, Morrison J H, Bloom F E, Sutcliffe J G
Department of Molecular Biology, Research Institute of Scripps Clinic, La Jolla, CA 92037.
Neuropharmacology. 1987 Jul;26(7B):845-54. doi: 10.1016/0028-3908(87)90061-x.
Morphological heterogeneity of individual neurons in the mammalian brain must ultimately result from differences between cells in their profiles of gene expression. However, the degree to which neurons located in different regions of the brain express different sets of genes is not known. Using differential cDNA cloning procedures, including subtractive hybridization and differential colony screening, a quantitative analysis of RNAs with heterogeneous distributions in the telencephalon of the rat and Old World monkey has been performed. The results suggest that no species of RNA exist with a distribution specific to the hippocampus or neocortex of the rat with an abundance greater than 0.05%. Additionally, the results with an abundance of 0.05% or greater which is present in neocortex of the monkey but absent from the cerebellum suggest that only one species of mRNA exists. A Northern blot analysis of cDNA clones representing mRNAs present in the neocortex but absent from the cerebellum of the monkey is presented, establishing the effectiveness of subtractive hybridization between different regions of the brain for cDNA cloning. Also presented is in situ hybridization analysis of the cellular distributions in primate neocortex of two mRNAs. One of these RNAs, detected by clone 1B4 is present in cortical laminae V and VI, and exhibits a high degree of heterogeneity in the overall density of labelled cells.
哺乳动物大脑中单个神经元的形态异质性最终必定源于细胞间基因表达谱的差异。然而，位于大脑不同区域的神经元表达不同基因集的程度尚不清楚。利用差异cDNA克隆程序，包括消减杂交和差异菌落筛选，对大鼠和旧世界猴端脑中分布不均一的RNA进行了定量分析。结果表明，不存在丰度大于0.05%且特异地分布于大鼠海马体或新皮质的RNA种类。此外，在猴新皮质中存在而在小脑中不存在且丰度为0.05%或更高的结果表明，仅存在一种mRNA。本文展示了对代表猴新皮质中存在而小脑中不存在的mRNA的cDNA克隆进行的Northern印迹分析，证实了大脑不同区域间消减杂交用于cDNA克隆的有效性。本文还展示了对两种mRNA在灵长类新皮质中的细胞分布进行的原位杂交分析。其中一种RNA由克隆1B4检测到，存在于皮质第V和第VI层，并且在标记细胞的总体密度上表现出高度异质性。
