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.
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We thank Fiqri Dizar Khaidizar and Ryutaro Akiyama for critical reading of the manuscript. We are grateful to the Ministry of Education, Culture, Sports, Science and Technology (MEXT), the Uehara Memorial Foundation, the Nakajima Foundation and the Mochida Memorial Foundation for past and current support.
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