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通过对180个连续的单个斑马鱼胚胎进行高分辨率时间进程分析确定斑马鱼早期胚胎发育过程中的转录组动态变化。

Transcriptome dynamics in early zebrafish embryogenesis determined by high-resolution time course analysis of 180 successive, individual zebrafish embryos.

作者信息

Rauwerda Han, Pagano Johanna F B, de Leeuw Wim C, Ensink Wim, Nehrdich Ulrike, de Jong Mark, Jonker Martijs, Spaink Herman P, Breit Timo M

机构信息

RNA Biology & Applied Bioinformatics research group, Swammerdam Institute for Life Sciences, Faculty of Science, University of Amsterdam, Amsterdam, The Netherlands.

Institute Biology Leiden, Faculty of Science, Leiden University, Leiden, The Netherlands.

出版信息

BMC Genomics. 2017 Apr 11;18(1):287. doi: 10.1186/s12864-017-3672-z.

Abstract

BACKGROUND

Recently, much progress has been made in the field of gene-expression in early embryogenesis. However, the dynamic behaviour of transcriptomes in individual embryos has hardly been studied yet and the time points at which pools of embryos are collected are usually still quite far apart. Here, we present a high-resolution gene-expression time series with 180 individual zebrafish embryos, obtained from nine different spawns, developmentally ordered and profiled from late blastula to mid-gastrula stage. On average one embryo per minute was analysed. The focus was on identification and description of the transcriptome dynamics of the expressed genes in this embryonic stage, rather than to biologically interpret profiles in cellular processes and pathways.

RESULTS

In the late blastula to mid-gastrula stage, we found 6,734 genes being expressed with low variability and rather gradual changes. Ten types of dynamic behaviour were defined, such as genes with continuously increasing or decreasing expression, and all expressed genes were grouped into these types. Also, the exact expression starting and stopping points of several hundred genes during this developmental period could be pinpointed. Although the resolution of the experiment was so high, that we were able to clearly identify four known oscillating genes, no genes were observed with a peaking expression. Additionally, several genes showed expression at two or three distinct levels that strongly related to the spawn an embryo originated from.

CONCLUSION

Our unique experimental set-up of whole-transcriptome analysis of 180 individual embryos, provided an unparalleled in-depth insight into the dynamics of early zebrafish embryogenesis. The existence of a tightly regulated embryonic transcriptome program, even between individuals from different spawns is shown. We have made the expression profile of all genes available for domain experts. The fact that we were able to separate the different spawns by their gene-expression variance over all expressed genes, underlines the importance of spawn specificity, as well as the unexpectedly tight gene-expression regulation in early zebrafish embryogenesis.

摘要

背景

最近,早期胚胎发育中的基因表达领域取得了很大进展。然而,单个胚胎中转录组的动态行为几乎尚未得到研究,而且收集胚胎池的时间点通常仍相距甚远。在此,我们展示了一个包含180个斑马鱼个体胚胎的高分辨率基因表达时间序列,这些胚胎来自九次不同的产卵,按发育顺序排列,并从囊胚晚期到原肠胚中期进行了分析。平均每分钟分析一个胚胎。重点在于识别和描述该胚胎阶段中表达基因的转录组动态,而非从生物学角度解释细胞过程和信号通路中的表达谱。

结果

在囊胚晚期到原肠胚中期阶段,我们发现6734个基因以低变异性和较为渐进的变化方式表达。定义了十种动态行为类型,例如表达持续增加或减少的基因,所有表达基因都被归类到这些类型中。此外,还能精确确定数百个基因在这个发育时期的准确表达起始和终止点。尽管实验分辨率很高,我们能够清晰识别四个已知的振荡基因,但未观察到具有表达峰值的基因。另外,几个基因在两个或三个不同水平上表达,这与胚胎所源自的产卵密切相关。

结论

我们对180个个体胚胎进行全转录组分析的独特实验设置,为斑马鱼早期胚胎发育的动态过程提供了无与伦比的深入见解。结果表明,即使在来自不同产卵的个体之间,也存在严格调控的胚胎转录组程序。我们已将所有基因的表达谱提供给领域专家。我们能够通过所有表达基因的基因表达差异来区分不同的产卵,这一事实强调了产卵特异性的重要性,以及斑马鱼早期胚胎发育中出乎意料的严格基因表达调控。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/d4d4/5387192/40ad594c071d/12864_2017_3672_Fig1_HTML.jpg

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