|ZFIN ID: ZDB-PUB-170323-21|
Imaging early embryonic calcium activity with GCaMP6s transgenic zebrafish
Chen, J., Xia, L., Bruchas, M.R., Solnica-Krezel, L.
|Source:||Developmental Biology 430(2): 385-396 (Journal)|
|Registered Authors:||Chen, Jiakun, Solnica-Krezel, Lilianna|
|Keywords:||Nodal, calcium transients, dorsal forerunner cells, embryonic cleavages, gastrulation, β-catenin|
|PubMed:||28322738 Full text @ Dev. Biol.|
Chen, J., Xia, L., Bruchas, M.R., Solnica-Krezel, L. (2017) Imaging early embryonic calcium activity with GCaMP6s transgenic zebrafish. Developmental Biology. 430(2):385-396.
ABSTRACTIntracellular Ca2+ signaling regulates cellular activities during embryogenesis and in adult organisms. We generated stable Tg[βactin2:GCaMP6s]stl351 and Tg[ubi:GCaMP6s]stl352 transgenic lines that combine the ubiquitously-expressed Ca2+ indicator GCaMP6s with the transparent characteristics of zebrafish embryos to achieve superior in vivo Ca2+ imaging. Using the Tg[βactin2:GCaMP6s]stl351 line featuring strong GCaMP6s expression from cleavage through gastrula stages, we detected higher frequency of Ca2+ transients in the superficial blastomeres during the blastula stages preceding the midblastula transition. Additionally, GCaMP6s also revealed that dorsal-biased Ca2+ signaling that follows the midblastula transition persisted longer during gastrulation, compared with earlier studies. We observed that dorsal-biased Ca2+ signaling is diminished in ventralized ichabod/β-catenin2 mutant embryos and ectopically induced in embryos dorsalized by excess β-catenin. During gastrulation, we directly visualized Ca2+ signaling in the dorsal forerunner cells, which form in a Nodal signaling dependent manner and later give rise to the laterality organ. We found that excess Nodal increases the number and the duration of Ca2+ transients specifically in the dorsal forerunner cells. The GCaMP6s transgenic lines described here enable unprecedented visualization of dynamic Ca2+ events from embryogenesis through adulthood, augmenting the zebrafish toolbox.