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Tracking neural crest cell cycle progression in vivo
Author(s) -
Rajan Sriivatsan G.,
Gallik Kristin L.,
Monaghan James R.,
Uribe Rosa A.,
Bronner Marianne E.,
Saxena Ankur
Publication year - 2018
Publication title -
genesis
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.093
H-Index - 110
eISSN - 1526-968X
pISSN - 1526-954X
DOI - 10.1002/dvg.23214
Subject(s) - neural crest , biology , zebrafish , population , cell cycle , microbiology and biotechnology , mcherry , live cell imaging , cell , genetics , green fluorescent protein , embryo , gene , demography , sociology
Analysis of cell cycle entry/exit and progression can provide fundamental insights into stem cell propagation, maintenance, and differentiation. The neural crest is a unique stem cell population in vertebrate embryos that undergoes long‐distance collective migration and differentiation into a wide variety of derivatives. Using traditional techniques such as immunohistochemistry to track cell cycle changes in such a dynamic population is challenging, as static time points provide an incomplete spatiotemporal picture. In contrast, the fluorescent, ubiquitination‐based cell cycle indicator (Fucci) system provides in vivo readouts of cell cycle progression and has been previously adapted for use in zebrafish. The most commonly used Fucci systems are ubiquitously expressed, making tracking of a specific cell population challenging. Therefore, we generated a transgenic zebrafish line, Tg(‐4.9sox10:mAG‐gmnn(1/100)‐2A‐mCherry‐cdt1(1/190)) , in which the Fucci system is specifically expressed in delaminating and migrating neural crest cells. Here, we demonstrate validation of this new tool and its use in live high‐resolution tracking of cell cycle progression in the neural crest and derivative populations.