Subtractive cloning of complementary DNAs and analysis of messenger RNAs with regional heterogeneous distributions in primate cortex.
Subtractive cloning of complementary DNAs and analysis of messenger RNAs with regional heterogeneous distributions in primate cortex.
复制标题
灵长类皮质中互补 DNA 的消减克隆和区域异质分布的信使 RNA 的分析。
DOI：
10.1016/0028-3908(87)90061-x
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发表时间：
1987
影响因子：
4.7
通讯作者：
Sutcliffe,JG
中科院分区：
文献类型：
--
作者：
Travis,GH;Naus,CG;Morrison,JH;Bloom,FE;Sutcliffe,JG
哺乳动物大脑中单个神经元的形态异质性最终必然是由细胞间基因表达谱的差异引起的。然而，位于大脑不同区域的神经元表达不同基因组的程度尚不清楚。利用差异cDNA克隆技术，包括消减杂交和差异集落筛选，对大鼠和旧大陆猴端脑中具有异质性分布的RNA进行了定量分析。结果表明，不存在丰度大于0.05%的对大鼠海马或新皮层具有特异性分布的RNA种类。另外，在猴的新皮质中存在的mRNA丰度为0.05%或更高，但小脑中不存在，这表明只有一种mRNA存在。对代表猴新皮质中存在但小脑中不存在的mRNA的cDNA克隆进行了北方印迹分析，建立了脑不同区域之间的消减杂交用于cDNA克隆的有效性。还对两种mRNA在灵长类动物大脑新皮层的细胞分布进行了原位杂交分析。这些RNA之一，检测克隆1B4是存在于皮质层V和VI，并表现出高度的异质性标记的细胞的整体密度。
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 isin situhybridization 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.