Analysis of Area-Specific Expression Patterns of RORbeta, ER81 and Nurr1 mRNAs in Rat Neocortex by Double In Situ Hybridization and Cortical Box Method
Analysis of Area-Specific Expression Patterns of RORbeta, ER81 and Nurr1 mRNAs in Rat Neocortex by Double In Situ Hybridization and Cortical Box Method
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DOI：
10.1371/journal.pone.0003266
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发表时间：
2008-09-25
期刊：
影响因子：
3.7
通讯作者：
Yamamori, Tetsuo
中科院分区：
文献类型：
--
作者：
Hirokawa, Junya;Watakabe, Akiya;Yamamori, Tetsuo
背景：哺乳动物的新皮层被细分为许多区域，每个区域都表现出独特的层状结构。为了详细研究这种区域差异，我们选择了三个基因进行比较分析，即RORbeta， ER81和Nurr1，它们的mrna分别主要表达在第4层，第5层和第6层。为了分析它们在大鼠新皮层中的定性和定量共表达谱，我们使用了我们之前开发的双原位杂交（ISH）组织化学和皮质盒法，将不同染色和个体的数据整合在一个标准的三维空间中。主要发现：我们的新方法产生了三个主要观察结果。首先，这三个基因表现出独特的区域分布模式，它们大多是互补的。皮质盒法显示的模式与尼氏染色确定的细胞结构区吻合良好。其次，在单细胞水平，RORbeta和ER81 mrna在第5层神经元亚群中共表达，而Nurr1和ER81 mrna没有共定位。第三，主成分分析表明，皮层的层次加工顺序与这三个基因的表达谱密切相关。在此基础上，我们认为体感区的非颗粒区（DZ）表现出一个更高阶的区域，这与之前的建议是一致的。结论/意义：揭示了三层特异性基因的表达与功能区域之间的密切关系，证明了皮质盒法在大脑皮层研究中的实用性。特别是，它使我们能够进行统计评估和模式匹配，这对于解释皮层中不断增加的基因表达数据将变得重要。
Background: The mammalian neocortex is subdivided into many areas, each of which exhibits distinctive lamina architecture. To investigate such area differences in detail, we chose three genes for comparative analyses, namely, RORbeta, ER81 and Nurr1, mRNAs of which have been reported to be mainly expressed in layers 4, 5 and 6, respectively. To analyze their qualitative and quantitative coexpression profiles in the rat neocortex, we used double in situ hybridization (ISH) histochemistry and cortical box method which we previously developed to integrate the data of different staining and individuals in a standard three-dimensional space.Principal Findings: Our new approach resulted in three main observations. First, the three genes showed unique area distribution patterns that are mostly complementary to one another. The patterns revealed by cortical box method matched well with the cytoarchitectonic areas defined by Nissl staining. Second, at single cell level, RORbeta and ER81 mRNAs were coexpressed in a subpopulation of layer 5 neurons, whereas Nurr1 and ER81 mRNAs were not colocalized. Third, principal component analysis showed that the order of hierarchical processing in the cortex correlates well with the expression profiles of these three genes. Based on this analysis, the dysgranular zone (DZ) in the somatosensory area was considered to exhibit a profile of a higher order area, which is consistent with previous proposal.Conclusions/Significance: The tight relationship between the expression of the three layer specific genes and functional areas were revealed, demonstrating the usefulness of cortical box method in the study on the cerebral cortex. In particular, it allowed us to perform statistical evaluation and pattern matching, which would become important in interpreting the ever-increasing data of gene expression in the cortex.