Determination of mRNA and protein expression patterns in zebrafish
- Christie, E.L., Parslow, A.C., and Heath, J.K.
- Methods in molecular biology (Clifton, N.J.) 469: 253-272 (Chapter)
- Registered Authors
- Christie, Elizabeth, Heath, Joan K., Parslow, Adam
- whole-mount in situ hybridization, riboprobe, whole-mount immunocytochemistry, antibody, vibrating microtome, sections
- MeSH Terms
- Gene Expression*
- Oligonucleotides, Antisense/genetics
- Oligonucleotides, Antisense/metabolism
- Signal Transduction/physiology
- Wnt Proteins/antagonists & inhibitors
- Wnt Proteins/metabolism
- Zebrafish/anatomy & histology
- Zebrafish/growth & development
- Zebrafish Proteins/genetics
- Zebrafish Proteins/metabolism
- 19109715 Full text @ Meth. Mol. Biol.
Christie, E.L., Parslow, A.C., and Heath, J.K. (2008) Determination of mRNA and protein expression patterns in zebrafish. Methods in molecular biology (Clifton, N.J.). 469:253-272.
Optically transparent zebrafish embryos provide an excellent vertebrate model system in which to reveal specific mRNA and protein expression patterns during development. Whole-mount preparations can be used to generate three-dimensional color or fluorescent readouts of the expression pattern of a given gene (or genes), matched with a bright-field image of all the tissues in the developing embryo. Whole-mount mRNA in situhybridization (WISH) has long been the method of choice for revealing gene expression patterns in zebrafish because this method depends only on being able to identify a relatively short region of nucleotide sequence unique for the gene of interest. In contrast, the scarcity of antibodies that are specific to or cross-react with zebrafish proteins has limited the widespread use of immunocyto-chemical applications, though this situation will improve in the future. The elucidation of the specific expression patterns of Wnt pathway genes in zebrafish has made a major contribution to our current understanding of their roles in vertebrate development.
Genes / Markers
Mutations / Transgenics
Human Disease / Model
Sequence Targeting Reagents
Engineered Foreign Genes