RNA <i>In Situ</i> Hybridization for Detecting Gene Expression Patterns in the Abdomens and Wings of <i>Drosophila</i> Species

Abstract

RNA in situ hybridization (ISH) is used to visualize spatio-temporal gene expression patterns with broad applications in biology and biomedicine. Here we provide a protocol for mRNA ISH in developing pupal wings and abdomens for model and non-model Drosophila species. We describe best practices in pupal staging, tissue preparation, probe design and synthesis, imaging of gene expression patterns, and image-editing techniques. This protocol has been successfully used to investigate the roles of genes underlying the evolution of novel color patterns in non-model Drosophila species.

Keywords

• <i>Drosophila</i>
• <i>in situ</i> hybridization
• gene expression