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在小鼠胚胎原肠胚形成过程中对振荡信号动力学的成像。

Imaging the onset of oscillatory signaling dynamics during mouse embryo gastrulation.

机构信息

Developmental Biology Unit, European Molecular Biology Laboratory, 69117 Heidelberg, Germany.

Division of Cell Biology, MRC Laboratory of Molecular Biology, Cambridge CB2 0QH, UK.

出版信息

Development. 2022 Jul 1;149(13). doi: 10.1242/dev.200083. Epub 2022 Jul 8.

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.

摘要

胚胎发育的一个基本要求是在空间和时间上协调信号活动。脊椎动物胚胎中一个显著的例子是体节形成过程中,与分段时钟相关的基因表达振荡在体节前中胚层 (PSM) 的细胞中被同步,导致组织水平的波状模式。为了在小鼠胚胎发育过程中研究它们的起始,我们研究了 Lfng 基因在原肠胚形成过程中的动力学。为此,我们建立了一个使用选择性平面照明显微镜 (SPIM) 的成像设置,该设置允许同时对多达四个胚胎进行培养和成像(“SPIM- for-4”)。使用 SPIM-for-4,结合遗传编码的信号报告基因,我们在早期形成的中胚层中检测到了 Lfng 振荡的起始,此时处于前体节阶段。功能上,我们发现即使 Notch 信号受到干扰,初始同步和前 6-8 个振荡周期也会发生,这表明与之前在斑马鱼胚胎中发现的结果相似。最后,我们表明在振荡活动开始时存在空间周期梯度,为我们的观察结果提供了一种潜在的机制,即波状模式在前几个振荡周期中逐渐建立。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/1132/9340547/e9ecd23f7480/develop-149-200083-g1.jpg

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