PUBLICATION

Whole Mount In Situ Hybridization and Immunohistochemistry for Zebrafish Larvae

Authors
Cunningham, R.L., Monk, K.R.
ID
ZDB-PUB-180317-9
Date
2018
Source
Methods in molecular biology (Clifton, N.J.)   1739: 371-384 (Chapter)
Registered Authors
Monk, Kelly
Keywords
Antibody, Fluorescence, Immunohistochemistry, In situ hybridization, RNA
MeSH Terms
  • Animals
  • Fluorescence
  • Immunohistochemistry/methods*
  • In Situ Hybridization/methods*
  • Larva/metabolism*
  • Motor Neurons/metabolism
  • RNA/metabolism
  • Zebrafish/metabolism*
PubMed
29546721 Full text @ Meth. Mol. Biol.
Abstract
In situ hybridization enables visualization of mRNA localization, and immunohistochemistry enables visualization of protein localization within a tissue or organism. Both techniques have been extensively utilized in zebrafish (Thisse et al., Development 119:1203-1215, 1993; Dutton et al., Development 128:4113-4125, 2001; Gilmour et al., Neuron 34:577-588, 2002; Lyons et al., Curr Biol 15:513-524, 2005) including for visualization of mRNA localization in Schwann cells (Lyons et al., Curr Biol 15:513-524, 2005; Monk et al., Science 325:1402-1405, 2009). For in situ hybridization, here, we outline how to generate RNA probes, conduct whole mount in situ hybridization for larvae, and list RNA probes that label different stages of Schwann cell development in zebrafish. For immunohistochemistry, the protocol we outline can be used to mark Schwann cells of sensory and motor nerves to examine properties such as developmental stage, morphology, proliferation, and apoptosis.
Genes / Markers
Figures
Expression
Phenotype
Mutations / Transgenics
Human Disease / Model
Sequence Targeting Reagents
Fish
Antibodies
Orthology
Engineered Foreign Genes
Mapping