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Methods Mol Biol. 2017;1487:367-378.

Analyzing ERK Signal Dynamics During Zebrafish Somitogenesis.

Author information

1
Gene Regulation Research, Nara Institute of Science and Technology, 8916-5, Takayama, Nara, 630-0101, Japan. matsui@bs.naist.jp.
2
Gene Regulation Research, Nara Institute of Science and Technology, 8916-5, Takayama, Nara, 630-0101, Japan.

Abstract

During vertebrate development, Erk is activated and regulates multiple cellular processes such as cell growth, differentiation, migration, and adhesion in a spatiotemporal manner. Whole-mount immunohistochemistry using antibodies against diphosphorylated Erk (p-Erk; active form of Erk) is a very useful method for understanding the spatial and temporal patterns of Erk activity during embryonic development. However, the fixation step of this method stops embryo development at a certain time point, making it very difficult to observe and interpret Erk activity dynamics. In this chapter, we describe a strategy that combines immunohistochemistry and quantitative analyses of multiple fixed embryos to reconstruct Erk activity dynamics during zebrafish somitogenesis.

KEYWORDS:

FGF; Immunohistochemistry; MAPK; Quantitative analysis; Somite; Zebrafish

PMID:
27924581
DOI:
10.1007/978-1-4939-6424-6_27
[Indexed for MEDLINE]

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