Multi-target Chromogenic Whole-mount In Situ Hybridization for Comparing Gene Expression Domains in Drosophila Embryos
2016 (English)In: Journal of Visualized Experiments, ISSN 1940-087X, E-ISSN 1940-087X, no 107, e53830Article in journal (Refereed) Published
To analyze gene regulatory networks active during embryonic development and organogenesis it is essential to precisely define how the different genes are expressed in spatial relation to each other in situ. Multi-target chromogenic whole-mount in situ hybridization (MC-WISH) greatly facilitates the instant comparison of gene expression patterns, as it allows distinctive visualization of different mRNA species in contrasting colors in the same sample specimen. This provides the possibility to relate gene expression domains topographically to each other with high accuracy and to define unique and overlapping expression sites. In the presented protocol, we describe a MC-WISH procedure for comparing mRNA expression patterns of different genes in Drosophila embryos. Up to three RNA probes, each specific for another gene and labeled by a different hapten, are simultaneously hybridized to the embryo samples and subsequently detected by alkaline phosphatase-based colorimetric immunohistochemistry. The described procedure is detailed here for Drosophila, but works equally well with zebrafish embryos.
Place, publisher, year, edition, pages
2016. no 107, e53830
Developmental Biology, Issue 107, Digoxigenin, biotin, fluorescein, azo dye, Fast Blue, Fast Red, INT, alkaline phosphatase substrate, WISH, fruit fly
Research subject Molecular Biology
IdentifiersURN: urn:nbn:se:su:diva-128914DOI: 10.3791/53830OAI: oai:DiVA.org:su-128914DiVA: diva2:917730