Koji T, Nakane P K
Department of Anatomy (III), Nagasaki University School of Medicine, Japan.
J Electron Microsc (Tokyo). 1996 Apr;45(2):119-27. doi: 10.1093/oxfordjournals.jmicro.a023420.
Over the past decade, considerable efforts to understand the states of specific gene expression at cellular and/or subcellular levels have been made. For this particular purpose, nonradioactive in situ hybridization to localize mRNAs has been developed and improved substantially, and it is now recognized as a powerful, established light-microscopical technique. In this review, we focus on the recent advances in the technological aspects of nonradioactive in situ hybridization including the use of synthetic oligodeoxynucleotide probes, the progress in analysis of signals, and the application to electron microscopy. Also, southwestern histochemistry, a relatively new method of localizing transcription regulatory proteins by utilizing haptenized DNA with responsive element sequences is described. Then we discuss what we can see by combining these molecular histochemical methods which were brought about by the merger of molecular biology and structural biology.
在过去十年中,人们付出了巨大努力来了解细胞和/或亚细胞水平上特定基因表达的状态。为了实现这一特定目的,用于定位mRNA的非放射性原位杂交技术得到了显著发展和改进,如今它已被公认为一种强大的、成熟的光学显微镜技术。在这篇综述中,我们重点关注非放射性原位杂交技术方面的最新进展,包括合成寡脱氧核苷酸探针的使用、信号分析的进展以及在电子显微镜中的应用。此外,还介绍了西南组织化学,这是一种通过利用带有反应元件序列的半抗原化DNA来定位转录调节蛋白的相对较新的方法。然后,我们讨论通过结合分子生物学和结构生物学融合带来的这些分子组织化学方法,我们能够看到什么。