Time-lapse observation of stepwise regression of Erk activity in zebrafish presomitic mesoderm.
Time-lapse observation of stepwise regression of Erk activity in zebrafish presomitic mesoderm.
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DOI：
10.1038/s41598-018-22619-9
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发表时间：
2018-03-12
影响因子：
4.6
通讯作者：
Matsui T
中科院分区：
文献类型：
--
作者：
Sari DWK;Akiyama R;Naoki H;Ishijima H;Bessho Y;Matsui T
在体节分裂过程中，时钟基因在后胚胎前中胚层(PSM)内振荡。时间信息与向后移动的成纤维细胞生长因子梯度联系在一起，导致在PSM内形成推定的体节。我们以前通过收集染色斑马鱼胚胎来研究成纤维细胞生长因子信号下游的Erk活性，发现Erk活性的陡峭梯度发生在PSM中，并且Erk活性边界以一种有规律的步进方式移动。然而，由于这些解释来自静态分析，我们需要通过应用具有更高时空分辨率的分析来坚定地确认它们。在这里，我们开发了一个斑马鱼胚胎中Erk活性的实时成像系统，使用基于Förster共振能量转移(FRET)的Erk生物传感器。通过这个系统，我们坚定地表明，ERK活性在PSM内呈现逐步回归。虽然我们的静态分析不能检测到时钟缺陷胚胎中ERK活性的逐步模式，但我们的系统显示，在时钟缺陷胚胎中，ERK活性的逐步回归发生在一个不规则的时间段，最终导致不规则大小的体节的形成。因此，我们的系统克服了静态分析的局限性，揭示了斑马鱼正常的体细胞发生需要ERK的时钟依赖的时空调节。
During somite segmentation, clock genes oscillate within the posterior presomitic mesoderm (PSM). The temporal information ties up with the posteriorly moving FGF gradient, leading to the formation of a presumptive somite within the PSM. We previously investigated Erk activity downstream of FGF signaling by collecting stained zebrafish embryos, and discovered that the steep gradient of Erk activity was generated in the PSM, and the Erk activity border regularly shifted in a stepwise manner. However, since these interpretations come from static analyses, we needed to firmly confirm them by applying an analysis that has higher spatiotemporal resolutions. Here we developed a live imaging system for Erk activity in zebrafish embryos, using a Förster resonance energy transfer (FRET)-based Erk biosensor. With this system, we firmly showed that Erk activity exhibits stepwise regression within the PSM. Although our static analyses could not detect the stepwise pattern of Erk activity in clock-deficient embryos, our system revealed that, in clock-deficient embryos, the stepwise regression of Erk activity occurs at an irregular timing, eventually leading to formation of irregularly-sized somites. Therefore, our system overcame the limitation of static analyses and revealed that clock-dependent spatiotemporal regulation of Erk is required for proper somitogenesis in zebrafish.
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