Chapter 14 Whole-Mount in Situ Hybridization for the Detection of RNA in Caenorhabditis elegans Embryos

Abstract

Publisher Summary This chapter discusses the whole-mount in situ hybridization for the detection of RNA in Caenorhabditis elegans embryos. In situ hybridization to RNA is an effective tool for the analysis of gene expression during development. This technique is particularly important for Caenorhabditis elegans, as isolation of RNA from specific tissues or developmental stages is generally not possible in this organism. The availability of the complete cell lineage and the reproducibility of cell positions from one animal to the next allow RNA expression patterns to be analyzed at the level of individual cells. A number of in situ hybridization protocols have been developed for the detection of RNA in squashed, dissected, or sectioned tissues of C. elegans. More recently, protocols using whole-mount preparations of C. elegans have also been described. By preserving the three-dimensional structure of the specimen, whole-mount preparations facilitate the identification of specific cells and the analysis of complex expression patterns. This chapter describes a protocol for detection of RNA in whole-mount C. elegans embryos. This procedure is based on protocols for Drosophila, which makes use of highly sensitive digoxigenin-labeled probes.