RNA in situ hybridization on planarian paraffin sections

Abstract

RNA in situ hybridization techniques are an important tool for the study of gene expression patterns in freshwater planarians. Here I describe a RNA in situ hybridization method on histological paraffin sections of planarian tissue. This protocol allows the visualization of gene expression at cellular or subcellular resolution. Following paraffin-embedding and sectioning of planarians, the resulting sections are hybridized with hapten-labeled RNA probes. Subsequent immunological probe detection is carried out with either chromogenic or fluorescent development. This protocol can be performed alone, or in combination with other immunodetection techniques, and represents a useful alternative to whole-mount protocols more commonly used in the community.