TY - JOUR
T1 - Neural Crest Methodologies in Zebrafish and Medaka
AU - Petratou, Kleio
AU - Camargo-Sosa, Karen
AU - Al Jabri, Ruqaiya
AU - Nagao, Yusuke
AU - Kelsh, Robert Neil
PY - 2019
Y1 - 2019
N2 - Neural crest cells are highly multipotent and strongly migratory cells and generate adult neural crest stem cells with varied roles in cellular homeostasis and regeneration. The optical transparency and ready accessibility of fish embryos make them particularly well-suited to high-resolution analysis of neural crest development. However, the dispersive nature of these cells adds to the challenge of their study, requiring that they be identified using marker expression. We describe key protocols for the analysis of neural crest marker expression in zebrafish and medaka, including whole-mount in situ hybridization to detect mRNA using conventional chromogenic substrates and the more recent RNAscope which gives readily multiplexed fluorescent detection and immunofluorescent detection of antigens.
AB - Neural crest cells are highly multipotent and strongly migratory cells and generate adult neural crest stem cells with varied roles in cellular homeostasis and regeneration. The optical transparency and ready accessibility of fish embryos make them particularly well-suited to high-resolution analysis of neural crest development. However, the dispersive nature of these cells adds to the challenge of their study, requiring that they be identified using marker expression. We describe key protocols for the analysis of neural crest marker expression in zebrafish and medaka, including whole-mount in situ hybridization to detect mRNA using conventional chromogenic substrates and the more recent RNAscope which gives readily multiplexed fluorescent detection and immunofluorescent detection of antigens.
KW - Animals
KW - Cell Movement/physiology
KW - Gene Expression Regulation, Developmental/physiology
KW - In Situ Hybridization
KW - Neural Crest/cytology
KW - Oryzias
KW - Zebrafish
UR - http://www.scopus.com/inward/record.url?scp=85064840454&partnerID=8YFLogxK
UR - http://www.scopus.com/inward/citedby.url?scp=85064840454&partnerID=8YFLogxK
U2 - 10.1007/978-1-4939-9412-0_13
DO - 10.1007/978-1-4939-9412-0_13
M3 - Article
C2 - 30977073
SN - 1064-3745
VL - 1976
SP - 167
EP - 184
JO - Methods in molecular biology (Clifton, N.J.)
JF - Methods in molecular biology (Clifton, N.J.)
ER -