In situ fluorescent RNA hybridization in insect small organs
Author of the article:CHEN Min** TANG Wen-Qian SHEN Jie WANG Dan***
Author's Workplace：Department of Entomology, China Agriculture University, Beijing 100193, China
Key Words：wing disc, RNA in situ hybridization, gene expression and regulation, trithorax, optomotor-blind
Abstract：[Objectives] RNA in situ hybridization is a popular technique for analyzing the expression and function of insect genes. This technique provides both qualitative and quantitative data on not only the spatio-temporal expression patterns of genes, but also on their cellular regulation. To optimize this technique, we performed RNA in situ fluorescent hybridization assays using the Drosophila larval wing disc as a representative insect small organ. [Methods] Wing discs of 3rd instar Drosophila larvae were dissected and used in RNA in situ hybridization. [Results] Many factors can influence the results of in situ hybridization, including the developmental stage of insect organs, probes, time and temperature of pre-hybridization and hybridization, washing time, and proper controls. We found that the cell memory gene trithorax is ubiquitously expressed in the wing disc and negatively regulated by the transcription factor Optomotor-blind. [Conclusion] In situ RNA hybridization is a convenient tool for revealing the expression patterns and regulation of genes in small organs of insects.