Plouhinec Jean-Louis, Medina-Ruiz Sofía, Borday Caroline, Bernard Elsa, Vert Jean-Philippe, Eisen Michael B, Harland Richard M, Monsoro-Burq Anne H
Université Paris Sud, Université Paris Saclay, CNRS UMR 3347, INSERM U1021, Orsay, France.
Institut Curie Research Division, PSL Research University, CNRS UMR 3347, INSERM U1021, Orsay, France.
PLoS Biol. 2017 Oct 19;15(10):e2004045. doi: 10.1371/journal.pbio.2004045. eCollection 2017 Oct.
During vertebrate neurulation, the embryonic ectoderm is patterned into lineage progenitors for neural plate, neural crest, placodes and epidermis. Here, we use Xenopus laevis embryos to analyze the spatial and temporal transcriptome of distinct ectodermal domains in the course of neurulation, during the establishment of cell lineages. In order to define the transcriptome of small groups of cells from a single germ layer and to retain spatial information, dorsal and ventral ectoderm was subdivided along the anterior-posterior and medial-lateral axes by microdissections. Principal component analysis on the transcriptomes of these ectoderm fragments primarily identifies embryonic axes and temporal dynamics. This provides a genetic code to define positional information of any ectoderm sample along the anterior-posterior and dorsal-ventral axes directly from its transcriptome. In parallel, we use nonnegative matrix factorization to predict enhanced gene expression maps onto early and mid-neurula embryos, and specific signatures for each ectoderm area. The clustering of spatial and temporal datasets allowed detection of multiple biologically relevant groups (e.g., Wnt signaling, neural crest development, sensory placode specification, ciliogenesis, germ layer specification). We provide an interactive network interface, EctoMap, for exploring synexpression relationships among genes expressed in the neurula, and suggest several strategies to use this comprehensive dataset to address questions in developmental biology as well as stem cell or cancer research.
在脊椎动物神经胚形成过程中,胚胎外胚层被分化为神经板、神经嵴、基板和表皮的谱系祖细胞。在这里,我们利用非洲爪蟾胚胎来分析神经胚形成过程中不同外胚层区域在细胞谱系建立期间的时空转录组。为了定义来自单个胚层的小细胞群的转录组并保留空间信息,通过显微切割将背侧和腹侧外胚层沿前后轴和内外侧轴进行细分。对这些外胚层片段的转录组进行主成分分析主要可识别胚胎轴和时间动态。这提供了一种遗传密码,可直接从其转录组定义任何外胚层样本沿前后轴和背腹轴的位置信息。同时,我们使用非负矩阵分解来预测早期和中期神经胚胚胎上增强的基因表达图谱以及每个外胚层区域的特定特征。时空数据集的聚类允许检测多个生物学相关组(例如,Wnt信号传导、神经嵴发育、感觉基板特化、纤毛发生、胚层特化)。我们提供了一个交互式网络界面EctoMap,用于探索神经胚中表达基因之间的共表达关系,并提出了几种利用这个综合数据集来解决发育生物学以及干细胞或癌症研究问题的策略。