Next-generation sequencing facilitates metal homeostasis and intoxication studies. Drosophila melanogaster

Purpose: Analyze transcriptome changes in response to metal treatment in wildtype and sensitized genetic backgrounds. Methods: Zinc or manganese chloride supplementation of wildtype or mutant Drosophila S2R+ cultured cells, followed by RNA extraction. Results: Zinc chloride supplementation induces transcriptional responses in wildtype, ia-2 mutant, and ZnT63C mutant cells. Conclusions: Wildtype S2R+ cells have a transcriptional response to mild zinc chloride supplementation and the zinc response is greater (in degree and in number of genes) in genetically sensitized cell backgrounds that perturb zinc mobilization. Overall design: The mRNA profiles of wild type or mutant S2R+ cells treated with 1mM ZnCl2 or MnCl2 for 24 hours, or untreated control cells, were generated by deep sequencing, in replicate, using Illumina HiSeq2000.研究目的：分析野生型与致敏遗传背景细胞经金属处理后产生的转录组变化。 研究方法：向野生型或突变型果蝇S2R+培养细胞中添加氯化锌或氯化锰，随后进行RNA提取。 研究结果：氯化锌处理可诱导野生型、ia-2突变型及ZnT63C突变型细胞产生转录应答。 研究结论：野生型S2R+细胞对轻度氯化锌处理存在转录应答；且在扰动锌动员的遗传致敏细胞背景中，锌应答的强度与涉及的基因数量均更为显著。 整体实验设计：分别对野生型或突变型S2R+细胞施加1mM氯化锌或氯化锰处理24小时，同时设置未处理对照组；通过Illumina HiSeq2000平台开展带有生物学重复的深度测序，以获取各组细胞的mRNA表达谱。