Whole Mount In Situ Hybridization and Immunohistochemistry for Zebrafish Larvae

Rebecca L Cunningham; Kelly R Monk

doi:10.1007/978-1-4939-7649-2_25

Whole Mount In Situ Hybridization and Immunohistochemistry for Zebrafish Larvae

Methods Mol Biol. 2018:1739:371-384. doi: 10.1007/978-1-4939-7649-2_25.

Authors

Rebecca L Cunningham¹, Kelly R Monk^{2

3}

Affiliations

¹ Department of Developmental Biology, Washington University in St. Louis, St. Louis, MO, USA.
² Department of Developmental Biology, Washington University in St. Louis, St. Louis, MO, USA. monk@ohsu.edu.
³ Vollum Institute, Oregon Health & Science University, Portland, OR, USA. monk@ohsu.edu.

PMID: 29546721
DOI: 10.1007/978-1-4939-7649-2_25

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.

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*

Substances

RNA