Effects of Larval Density on Gene Regulation in Caenorhabditis elegans During Routine L1 Synchronization.
Effects of Larval Density on Gene Regulation in Caenorhabditis elegans During Routine L1 Synchronization.
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DOI：
10.1534/g3.118.200056
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发表时间：
2018-05-04
期刊：
影响因子：
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通讯作者：
Conine CC
中科院分区：
文献类型：
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作者：
Chan IL;Rando OJ;Conine CC
漂白妊娠C.线虫，然后对L1幼虫进行短期饥饿是蠕虫研究人员为产生用于实验的同步种群而进行的常规方法。在利用单虫RNA-Seq技术研究饲料对L1期蠕虫基因调控的影响过程中，我们发现重悬L1期幼虫的密度影响许多mRNA的表达。具体而言，一些与代谢和信号传导相关的基因在低密度下被捕的蠕虫中高度表达，但在较高的被捕密度下被抑制。我们基于我们数据集中最依赖密度的基因之一- lips-15 -生成了GFP报告菌株，并证实该报告菌株在相对低密度下被捕获的蠕虫中特异性表达。最后，我们表明，条件培养基从高密度L1文化能够下调唇-15，即使在L1动物逮捕在低密度，和实验使用daf-22突变动物表明，这种效果是不介导的信号信息素的蛔虫苷家族。总之，我们的数据暗示了一种可溶性信号分子在L1阶段C的密度感知中。elegans，并提供指导的实验设计集中在早期发育基因调控。
Bleaching gravid C. elegans followed by a short period of starvation of the L1 larvae is a routine method performed by worm researchers for generating synchronous populations for experiments. During the process of investigating dietary effects on gene regulation in L1 stage worms by single-worm RNA-Seq, we found that the density of resuspended L1 larvae affects expression of many mRNAs. Specifically, a number of genes related to metabolism and signaling are highly expressed in worms arrested at low density, but are repressed at higher arrest densities. We generated a GFP reporter strain based on one of the most density-dependent genes in our dataset – lips-15 – and confirmed that this reporter was expressed specifically in worms arrested at relatively low density. Finally, we show that conditioned media from high density L1 cultures was able to downregulate lips-15 even in L1 animals arrested at low density, and experiments using daf-22 mutant animals demonstrated that this effect is not mediated by the ascaroside family of signaling pheromones. Together, our data implicate a soluble signaling molecule in density sensing by L1 stage C. elegans, and provide guidance for design of experiments focused on early developmental gene regulation.