Whole-Mount In Situ Hybridization in Large Sexual Schmidtea mediterranea.

Whole-mount in situ hybridization (WISH), colorimetric or fluorescent (FISH), allows for the visualization of endogenous RNA. For planarians, robust WISH protocols exist for small-sized animals (>5 mm) of the model species Schmidtea mediterranea and Dugesia japonica. However, the sexual strain of Schmidtea mediterranea studied for germline development and function reaches much larger body sizes in excess of 2 cm. The existing whole-mount WISH protocols are not optimal for such large specimens, owing to insufficient tissue permeabilization. Here, we describe a robust WISH protocol for 12-16 mm long sexually mature Schmidtea mediterranea individuals that could serve as a starting point for adapting WISH to other large planarian species.

Whole-Mount In Situ Hybridization of Planarians.

Whole-mount in situ hybridization (WISH) and fluorescent whole-mount in situ hybridization (FISH) allow for visualization of specific mRNA transcripts to answer diverse biological questions. In planarians, in situ hybridization enables determination of gene expression profiles and identification of cell-type specific markers for analyzing experimental treatments. Here, we describe a robust whole-mount protocol for detecting gene expression patterns in the planarian Schmidtea mediterranea.