Imaging early embryonic calcium activity with GCaMP6s transgenic zebrafish.
Imaging early embryonic calcium activity with GCaMP6s transgenic zebrafish.
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DOI：
10.1016/j.ydbio.2017.03.010
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发表时间：
2017-10-15
影响因子：
2.7
通讯作者：
Solnica-Krezel L
中科院分区：
文献类型：
--
作者：
Chen J;Xia L;Bruchas MR;Solnica-Krezel L
细胞内钙信号调节胚胎发育和成年生物体的细胞活动。我们建立了稳定的TG[βactin2：GCaMP6s]st1351和TG[ubi：GCaMP6s]st1352转基因系，将普遍表达的钙指示剂GCaMP6s与斑马鱼胚胎的透明特性相结合，实现了良好的体内钙成像。利用从卵裂到原肠胚高表达GCaMP6s的TG[βactin2：GCaMP6s]st1351细胞系，我们检测到在囊胚期之前的囊胚期，浅层囊胚球中钙离子瞬变的频率较高。此外，GCaMP6还表明，与以前的研究相比，伴随着中期囊胚转变的背侧偏向的钙信号在原肠形成过程中持续的时间更长。我们观察到，在内侧化的Ichabod/β-catenin2突变胚胎中，背侧偏向的钙信号减弱，而在过量的β-catenin背侧的胚胎中异位诱导。在原肠发育过程中，我们在背侧前体细胞中直接观察到了钙信号，这些细胞以结节信号依赖的方式形成，后来形成了偏侧器官。我们发现，过量的结节增加了钙瞬变的数量和持续时间，特别是在背侧前体细胞中。这里描述的GCaMP6s转基因株系能够前所未有地可视化从胚胎发生到成年的动态钙事件，增强了斑马鱼的工具箱。
Intracellular 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.
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影响因子：
56.9
作者：
De Koninck, P;Schulman, H
通讯作者：
Schulman, H
影响因子：
9.2
作者：
Amack, JD;Yost, HJ
通讯作者：
Yost, HJ
DOI：
10.1073/pnas.97.21.11403
发表时间：
2000-10-10
影响因子：
11.1
作者：
Kawakami, K;Shima, A;Kawakami, N
通讯作者：
Kawakami, N
影响因子：
64.5
作者：
JAENISCH, R;JAHNER, D;GROTKOPP, D
通讯作者：
GROTKOPP, D
影响因子：
7.8
作者：
CHANG, DC;MENG, CL
通讯作者：
MENG, CL