Non-radioactive detection of nerve growth factor receptor (NGFR) mRNA in rat brain using in situ hybridization histochemistry.
Non-radioactive detection of nerve growth factor receptor (NGFR) mRNA in rat brain using in situ hybridization histochemistry.
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DOI：
10.1177/39.2.1846159
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发表时间：
1991-02
期刊：
影响因子：
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通讯作者：
J. Springer;E. Robbins;B. Gwag;M. E. Lewis;F. Baldino
中科院分区：
文献类型：
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作者：
J. Springer;E. Robbins;B. Gwag;M. E. Lewis;F. Baldino
放射性标记的RNA探针结合原位杂交组织化学已成为研究中枢神经系统基因表达的有用方法。我们使用地高辛标记的尿苷三磷酸合成cRNA探针定位神经生长因子受体（NGFR）mRNA在大鼠基底前脑。使用与碱性磷酸酶偶联的地高辛抗体检测含有地高辛标记的NGFR mRNA的细胞。NGFR mRNA阳性细胞分布在基底前脑的三个主要细胞群：内侧隔核、Broca斜角带的垂直和水平分支以及基底核。该技术提供了一种快速和灵敏的方法，用于高分辨率检测中枢神经系统中的mRNA种类，以及两种不同mRNA种类在单个细胞内共定位的潜力。
Radioactively labeled RNA probes in conjunction with in situ hybridization histochemistry have become a useful method for studying gene expression in the central nervous system. We used digoxigenin-labeled uridine triphosphate to synthesize cRNA probes for localization of nerve growth factor receptor (NGFR) mRNA in the rat basal forebrain. Detection of cells containing digoxigenin-labeled NGFR mRNA was accomplished using a digoxigenin antibody conjugated with alkaline phosphatase. NGFR mRNA-positive cells were distributed in three major cell groups in the basal forebrain: the medial septal nucleus, vertical and horizontal limbs of the diagonal band of Broca, and nucleus basalis. This technique provides a rapid and sensitive method for high-resolution detection of mRNA species in the central nervous system, as well as the potential for co-localization of two different mRNA species within individual cells.