mRNA profiling reveals significant transcriptional differences between a multipotent progenitor and its differentiated sister
mRNA profiling reveals significant transcriptional differences between a multipotent progenitor and its differentiated sister
复制标题
DOI：
10.1186/s12864-019-5821-z
复制
发表时间：
2019-05-28
期刊：
影响因子：
4.4
通讯作者：
Bettinger, Jill C.
中科院分区：
文献类型：
--
作者：
Mathies, Laura D.;Ray, Surjyendu;Bettinger, Jill C.
研究背景秀丽隐杆线虫的两种体细胞性腺前体细胞（sogonadal precursors，SGPs）是发育成体生殖系统所有体细胞组织的多能祖细胞。SGPs的姐妹细胞是两个头部中胚层细胞（hmcs）;一个hmcs通过程序性细胞死亡而死亡，另一个终末分化。因此，单个细胞分裂产生一个多能祖细胞和一个具有相同谱系历史的分化细胞。我们比较了SGPs和hmcs的转录组，以了解在这个lineage.ResultsWe的多能性和分化的决定因素，产生了一个菌株，特别是在SGPs（ehn-3A：：tdTomato）和hmcs（bgal-1：：GFP）表达荧光标记。在SGP/hmc细胞分裂后，但在SGP进一步分裂之前，我们从动物中分离细胞，并将分离的细胞进行荧光激活细胞分选以收集分离的SGP和hmcs。我们分析了这些细胞的转录组，发现5912个转录物在两种细胞类型之间显著差异表达，表达变化至少为两倍。偏hmc基因富含那些神经元特有的基因。SGP-biased基因富集了那些指示细胞增殖和发育的基因。我们评估了我们的差异表达的基因的有效性，通过检查现有的报告的10个基因中的5个最显着的偏差在SGPs中的表达，发现两个显示在SGPs中的表达。对于一个在SGPs中未显示表达的报告基因，我们使用CRISPR/Cas9产生GFP敲入。这个报告，在本机的基因组背景下，在SGPs.ConclusionsWe发现，SGPs和hmcs的转录谱是惊人的不同。偏向hmc的基因富含那些编码突触传递机制的基因，这强烈表明它具有神经元样的信号传递特性。相反，SGP偏向基因富含编码参与转录和翻译的因子的基因，正如从准备进行增殖分裂的细胞中所预期的那样。多能性的介体可能是SGPs中差异表达的基因之一。
BackgroundThe two Caenorhabditis elegans somatic gonadal precursors (SGPs) are multipotent progenitors that generate all somatic tissues of the adult reproductive system. The sister cells of the SGPs are two head mesodermal cells (hmcs); one hmc dies by programmed cell death and the other terminally differentiates. Thus, a single cell division gives rise to one multipotent progenitor and one differentiated cell with identical lineage histories. We compared the transcriptomes of SGPs and hmcs in order to learn the determinants of multipotency and differentiation in this lineage.ResultsWe generated a strain that expressed fluorescent markers specifically in SGPs (ehn-3A::tdTomato) and hmcs (bgal-1::GFP). We dissociated cells from animals after the SGP/hmc cell division, but before the SGPs had further divided, and subjected the dissociated cells to fluorescence-activated cell sorting to collect isolated SGPs and hmcs. We analyzed the transcriptomes of these cells and found that 5912 transcripts were significantly differentially expressed, with at least two-fold change in expression, between the two cell types. The hmc-biased genes were enriched with those that are characteristic of neurons. The SGP-biased genes were enriched with those indicative of cell proliferation and development. We assessed the validity of our differentially expressed genes by examining existing reporters for five of the 10 genes with the most significantly biased expression in SGPs and found that two showed expression in SGPs. For one reporter that did not show expression in SGPs, we generated a GFP knock-in using CRISPR/Cas9. This reporter, in the native genomic context, was expressed in SGPs.ConclusionsWe found that the transcriptional profiles of SGPs and hmcs are strikingly different. The hmc-biased genes are enriched with those that encode synaptic transmission machinery, which strongly suggests that it has neuron-like signaling properties. In contrast, the SGP-biased genes are enriched with genes that encode factors involved in transcription and translation, as would be expected from a cell preparing to undergo proliferative divisions. Mediators of multipotency are likely to be among the genes differentially expressed in SGPs.