Yabe Tomio, Maeda Nobuaki
Faculty of Applied Biological Sciences, Department of Applied Life Science, Gifu University, 1-1 Yanagido, Gifu, 501-1193, Japan,
Methods Mol Biol. 2015;1229:377-87. doi: 10.1007/978-1-4939-1714-3_29.
In situ hybridization provides information for understanding the localization of gene expression in various tissues. The relative expression levels of mRNAs in a single cell can be sensitively visualized by this technique. Furthermore, since in situ hybridization is a histological technique, tissue structure is maintained after fixation, and it is possible to accurately identify cell types. We have examined the expression of heparan sulfate sulfotransferases by in situ hybridization to better understand the functions of heparan sulfate in the development of mouse nervous system. This chapter describes methods of in situ hybridization analyses using cRNA probes labeled with nonradioactive nucleotides.
原位杂交为理解基因在各种组织中的表达定位提供了信息。通过该技术可以灵敏地观察到单个细胞中mRNA的相对表达水平。此外,由于原位杂交是一种组织学技术,固定后组织结构得以保留,从而能够准确识别细胞类型。我们通过原位杂交检测了硫酸乙酰肝素硫酸转移酶的表达,以更好地了解硫酸乙酰肝素在小鼠神经系统发育中的功能。本章介绍了使用非放射性核苷酸标记的cRNA探针进行原位杂交分析的方法。