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ZFIN ID: ZDB-PUB-110429-1
Visualization of mRNA Expression in the Zebrafish Embryo
Machluf, Y., and Levkowitz, G.
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.
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.