小鼠胚胎原肠胚形成过程中振荡信号动力学的成像

Henning J. Falk, Takehito Tomita, Gregor Mönke, K. McDole, A. Aulehla
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引用次数: 10

摘要

胚胎发育的一个基本要求是信号活动在空间和时间上的协调。在脊椎动物胚胎中,一个显著的例子是在体细胞发育过程中发现的,在体细胞前中胚层(PSM)中,与分割时钟相关的基因表达振荡在细胞间同步,并导致组织水平的波模式。为了研究它们在小鼠胚胎发育过程中的发生,我们研究了分割时钟基因lng在原肠胚形成过程中的动态变化。为此,我们使用选择性平面照明显微镜(SPIM)建立了一种成像装置,可以培养和同时成像多达四个胚胎(' SPIM- for-4 ')。利用SPIM-for-4,结合遗传编码的信号报告,我们检测到了在前体细胞阶段新形成的中胚层中lfg振荡的开始。在功能上,我们发现即使Notch信号受损,初始同步和第一个~ 6-8个振荡周期也会发生,这与之前在斑马鱼胚胎中的发现相似。最后,我们表明在振荡活动开始时存在空间周期梯度,这为我们观察到的波形在第一个振荡周期中逐渐建立提供了一个潜在的机制。
本文章由计算机程序翻译,如有差异,请以英文原文为准。
Imaging the onset of oscillatory signaling dynamics during mouse embryo gastrulation
ABSTRACT A fundamental requirement for embryonic development is the coordination of signaling activities in space and time. A notable example in vertebrate embryos is found during somitogenesis, where gene expression oscillations linked to the segmentation clock are synchronized across cells in the presomitic mesoderm (PSM) and result in tissue-level wave patterns. To examine their onset during mouse embryo development, we studied the dynamics of the segmentation clock gene Lfng during gastrulation. To this end, we established an imaging setup using selective plane illumination microscopy (SPIM) that enables culture and simultaneous imaging of up to four embryos (‘SPIM- for-4’). Using SPIM-for-4, combined with genetically encoded signaling reporters, we detected the onset of Lfng oscillations within newly formed mesoderm at presomite stages. Functionally, we found that initial synchrony and the first ∼6-8 oscillation cycles occurred even when Notch signaling was impaired, revealing similarities to previous findings made in zebrafish embryos. Finally, we show that a spatial period gradient is present at the onset of oscillatory activity, providing a potential mechanism accounting for our observation that wave patterns build up gradually over the first oscillation cycles.
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