PUBLICATION
Visualization of mRNA Expression in the Zebrafish Embryo
- Authors
- Machluf, Y., and Levkowitz, G.
- ID
- ZDB-PUB-110429-1
- Date
- 2011
- Source
- Methods in molecular biology (Clifton, N.J.) 714: 83-102 (Chapter)
- Registered Authors
- Levkowitz, Gil, Machluf, Yossy
- Keywords
- none
- MeSH Terms
-
- Animals
- Antibodies/immunology
- Antigens/immunology
- Color
- Colorimetry
- Embryo, Nonmammalian/cytology
- Embryo, Nonmammalian/metabolism*
- Gene Expression Regulation*
- Immunohistochemistry
- In Situ Hybridization/methods*
- In Situ Hybridization, Fluorescence
- RNA Probes/genetics
- RNA, Messenger/analysis*
- RNA, Messenger/genetics
- RNA, Messenger/metabolism
- Zebrafish/embryology*
- PubMed
- 21431736 Full text @ Meth. Mol. Biol.
Citation
Machluf, Y., and Levkowitz, G. (2011) Visualization of mRNA Expression in the Zebrafish Embryo. Methods in molecular biology (Clifton, N.J.). 714:83-102.
Abstract
Examination of spatial and temporal gene expression pattern is a key step towards understanding gene function. Therefore, in situ hybridization of mRNA is one of the most powerful and widely used -techniques in biology. Recent advances allow the reliable and simultaneous detection of mRNA transcripts, or combinations of mRNA and protein, in zebrafish embryos.Here we describe a standard protocol for visualizing the precise expression pattern of a single transcript or multiple gene products. The procedure employs fixation and permeabilization of embryos, -followed by hybridization with tagged antisense riboprobes. Excess probes are then washed and hybrids are detected by enzyme-mediated immunohistochemistry utilizing either chromogenic or fluorescent substrates.
Genes / Markers
Expression
Phenotype
Mutations / Transgenics
Human Disease / Model
Sequence Targeting Reagents
Fish
Orthology
Engineered Foreign Genes
Mapping