Shih Nathan P, François Paul, Delaune Emilie A, Amacher Sharon L
Department of Molecular and Cell Biology, University of California, Berkeley, CA 94720, USA.
Department of Physics, McGill University, Montréal, Canada H3A 2T8.
Development. 2015 May 15;142(10):1785-93. doi: 10.1242/dev.119057.
The formation of reiterated somites along the vertebrate body axis is controlled by the segmentation clock, a molecular oscillator expressed within presomitic mesoderm (PSM) cells. Although PSM cells oscillate autonomously, they coordinate with neighboring cells to generate a sweeping wave of cyclic gene expression through the PSM that has a periodicity equal to that of somite formation. The velocity of each wave slows as it moves anteriorly through the PSM, although the dynamics of clock slowing have not been well characterized. Here, we investigate segmentation clock dynamics in the anterior PSM in developing zebrafish embryos using an in vivo clock reporter, her1:her1-venus. The her1:her1-venus reporter has single-cell resolution, allowing us to follow segmentation clock oscillations in individual cells in real-time. By retrospectively tracking oscillations of future somite boundary cells, we find that clock reporter signal increases in anterior PSM cells and that the periodicity of reporter oscillations slows to about ∼1.5 times the periodicity in posterior PSM cells. This gradual slowing of the clock in the anterior PSM creates peaks of clock expression that are separated at a two-segment periodicity both spatially and temporally, a phenomenon we observe in single cells and in tissue-wide analyses. These results differ from previous predictions that clock oscillations stop or are stabilized in the anterior PSM. Instead, PSM cells oscillate until they incorporate into somites. Our findings suggest that the segmentation clock may signal somite formation using a phase gradient with a two-somite periodicity.
沿脊椎动物身体轴重复形成体节是由体节时钟控制的,体节时钟是一种在前体节中胚层(PSM)细胞内表达的分子振荡器。尽管PSM细胞自主振荡,但它们与相邻细胞协调,通过PSM产生一波循环基因表达的扫波,其周期与体节形成的周期相等。每一波的速度在向前穿过PSM时都会减慢,尽管时钟减慢的动力学尚未得到很好的表征。在这里,我们使用体内时钟报告基因her1:her1-venus研究斑马鱼胚胎发育过程中前体节中胚层的体节时钟动力学。her1:her1-venus报告基因具有单细胞分辨率,使我们能够实时跟踪单个细胞中的体节时钟振荡。通过回顾性跟踪未来体节边界细胞的振荡,我们发现前体节中胚层细胞中的时钟报告信号增加,并且报告振荡的周期减慢到后体节中胚层细胞周期的约1.5倍。前体节中胚层中时钟的这种逐渐减慢在空间和时间上以两个体节的周期产生时钟表达峰值,这是我们在单细胞和全组织分析中观察到的一种现象。这些结果与之前关于时钟振荡在前体节中胚层中停止或稳定的预测不同。相反,PSM细胞振荡直到它们并入体节。我们的研究结果表明,体节时钟可能使用具有两个体节周期的相位梯度来指示体节形成。