Department of Physiology and Biophysics, University of Washington, Seattle, Washington 98195, USA.
Mol Cell Neurosci. 1992 Oct;3(5):395-405. doi: 10.1016/1044-7431(92)90051-3.
We present a new method for the simultaneous detection of two mRNA species within individual neurons. The technique involves the use of radio-labeled and digoxigenin-labeled cRNA probes, the application of which confers a high specificity and sensitivity to the in situ hybridization analysis. We demonstrate the use of this method by illustrating the coexpression of preprogonadotropin-releasing hormone (GnRH) mRNA and preprogalanin mRNA in neurons in the rat forebrain and report a distinct sexual dimorphism in galanin gene expression by GnRH neurons. Coupling this technology with semi-quantitative analysis of the mRNA species hybridized with the isotopically labeled mRNA would permit studies of gene regulation in individual cells among the heterogeneous populations of the brain.
我们提出了一种新方法，可用于在单个神经元中同时检测两种 mRNA 分子。该技术涉及放射性标记和地高辛标记的 cRNA 探针的应用，这为原位杂交分析提供了高度的特异性和敏感性。我们通过展示 GnRH 前体释放激素 (GnRH) mRNA 和前甘丙肽原 mRNA 在大鼠前脑中神经元中的共表达，证明了该方法的用途，并报告了 GnRH 神经元中甘丙肽基因表达的明显性别二态性。将这项技术与用同位素标记的 mRNA 杂交的 mRNA 种类的半定量分析相结合，将允许在大脑的异质细胞群体中研究单个细胞中的基因调控。
