TY - CHAP
T1 - Whole mount in situ hybridization and immunohistochemistry for zebrafish larvae
AU - Cunningham, Rebecca L.
AU - Monk, Kelly R.
N1 - Publisher Copyright:
© 2018, Springer Science+Business Media, LLC.
PY - 2018
Y1 - 2018
N2 - 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.
AB - 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.
KW - Antibody
KW - Fluorescence
KW - Immunohistochemistry
KW - In situ hybridization
KW - RNA
UR - http://www.scopus.com/inward/record.url?scp=85044182436&partnerID=8YFLogxK
UR - http://www.scopus.com/inward/citedby.url?scp=85044182436&partnerID=8YFLogxK
U2 - 10.1007/978-1-4939-7649-2_25
DO - 10.1007/978-1-4939-7649-2_25
M3 - Chapter
C2 - 29546721
AN - SCOPUS:85044182436
T3 - Methods in Molecular Biology
SP - 371
EP - 384
BT - Methods in Molecular Biology
PB - Humana Press Inc.
ER -