Effects of ambient temperature on transcript levels and alternative polyadenylation in Drosophila melanogaster [RNA-seq]

Cells in ectothermic organisms often maintain homeostatic function over a considerable range of ambient temperatures. However, as temperature has pronounced effects on all biological processes, but not necessarily in a uniform manner on each of the myriad of distinct processes, cellular acclimation to ambient temperature change is predicted to involve complex regulation. To assess the effects of temperature change within the readily tolerated range on the transcriptome, we have performed analyses with the ectothermic organism Drosophila melanogaster. Both adult male flies and S2R+ cells were analyzed. 3' RNA-Seq was applied to study effects of ambient temperature change on transcript levels and on polyadenylation site selection. Overall design: Aliquots of adult male flies and S2R+ cells, respectively, were shifted to distinct temperatures (11, 14, 25 and 30°C). Twenty-four hours later, total RNA was isolated and analyzed by 3' RNA-Seq. Three replicate experiments were performed.

变温生物（ectothermic organisms）体内的细胞，通常可在较宽的环境温度范围内维持稳态功能。然而，温度对所有生物学过程均具有显著影响，但对这海量不同生理过程的作用未必统一，因此推测细胞对环境温度变化的驯化过程涉及复杂的调控机制。为探究生物体易耐受温度范围内的温度变化对转录组（transcriptome）的影响，我们以变温生物黑腹果蝇（Drosophila melanogaster）为研究对象开展了相关分析。本研究同时对成年雄性果蝇与S2R+细胞进行了分析。我们采用3'端RNA测序（3' RNA-Seq）技术，旨在探究环境温度变化对转录本水平及多腺苷酸化位点选择的影响。实验整体设计：分别将成年雄性果蝇与S2R+细胞的等分试样转移至11、14、25及30℃这4个不同温度环境中培养。24小时后，提取总RNA并通过3'端RNA测序进行检测分析。本实验共设置3次生物学重复。