ERK Activity Dynamics during Zebrafish Embryonic Development
ERK Activity Dynamics during Zebrafish Embryonic Development
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DOI：
10.3390/ijms20010109
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发表时间：
2018-12
影响因子：
5.6
通讯作者：
Kah-Loon Wong;Ryutaro Akiyama;Yasumasa Bessho;Takaaki Matsui
中科院分区：
文献类型：
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作者：
Kah-Loon Wong;Ryutaro Akiyama;Yasumasa Bessho;Takaaki Matsui
在脊椎动物发育过程中，细胞外信号调节激酶（ERK）被生长因子如成纤维细胞生长因子（FGF）激活，并且其调节组织/器官（包括眼、脑、体节、四肢和内耳）的形成。然而，一个实验系统，以监测ERK活性动力学在整个身体的脊椎动物胚胎是缺乏的。我们最近研究了ERK活性动态的前体节中胚层的活斑马鱼胚胎注射编码的Förster共振能量转移（FRET）为基础的ERK生物传感器的mRNA。在这项研究中，转基因斑马鱼稳定和普遍表达的ERK生物传感器产生监测ERK活性动态整个胚胎发育。该系统允许识别胚胎中的ERK激活结构域从囊胚晚期到后期分割阶段，与使用抗磷酸化ERK抗体获得的免疫染色模式一致。产生了斑马鱼胚胎发生过程中整个身体的ERK活性的时空图，并鉴定了先前未鉴定的激活动力学和ERK结构域。该系统是第一个报告的方法来监测ERK活性动态在脊椎动物胚胎发育，提供洞察ERK活性的作用，在正常和异常发育的活脊椎动物胚胎。
During vertebrate development, extracellular signal-regulated kinase (ERK) is activated by growth factors such as fibroblast growth factor (FGF), and it regulates the formation of tissues/organs including eyes, brains, somites, limbs, and inner ears. However, an experimental system to monitor ERK activity dynamics in the entire body of the vertebrate embryo is lacking. We recently studied ERK activity dynamics in the pre-somitic mesoderm of living zebrafish embryos injected with mRNAs encoding a Förster resonance energy transfer (FRET)-based ERK biosensor. In this study, transgenic zebrafish stably and ubiquitously expressing the ERK biosensor were generated to monitor ERK activity dynamics throughout embryonic development. The system allowed the identification of ERK activation domains in embryos from the late blastula to the late segmentation stage, consistent with immunostaining patterns obtained using anti-phosphorylated ERK antibody. A spatiotemporal map of ERK activity in the entire body during zebrafish embryogenesis was generated, and previously unidentified activation dynamics and ERK domains were identified. The proposed system is the first reported method to monitor ERK activity dynamics during vertebrate embryogenesis, providing insight into the role of ERK activity in normal and abnormal development in living vertebrate embryos.