title: Multi-target Chromogenic Whole-mount In Situ Hybridization for Comparing Gene Expression Domains in Drosophila Embryos.
abstract: 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.
authors: Department of Molecular Biosciences, The Wenner-Gren Institute (MBW), Stockholm University; giselbert.hauptmann@su.se.
authors: Giselbert
authors: G
authors: Hauptmann
authors: Department of Molecular Biosciences, The Wenner-Gren Institute (MBW), Stockholm University.
authors: Iris
authors: I
authors: Söll
authors: Department of Molecular Biosciences, The Wenner-Gren Institute (MBW), Stockholm University.
authors: Robert
authors: R
authors: Krautz
authors: Department of Molecular Biosciences, The Wenner-Gren Institute (MBW), Stockholm University.
authors: Ulrich
authors: U
authors: Theopold
mesh_terms: D000818:Animals; D004330:Drosophila / Q000196:embryology* / Q000235:genetics*; D047108:Embryonic Development / Q000235:genetics; D015870:Gene Expression; D018507:Gene Expression Regulation, Developmental; D007150:Immunohistochemistry; D017403:In Situ Hybridization / Q000379:methods*; D015347:RNA Probes / Q000737:chemistry; D012333:RNA, Messenger / Q000378:metabolism
chemical_list: D015347:RNA Probes; D012333:RNA, Messenger