The impact of chronic temperature elevation on the spleen anti-viral transcriptomic response in Atlantic cod (Gadus morhua)

This experiment was designed to investigate the impacts of non-lethal increases in temperature on the anti-viral transcriptomic response of Atlantic cod. Selected genes identified as differentially expressed between samples injected with pIC but held at different temperatures were validated using QPCR. This was a common reference design. For the common reference each individual sample analyzed in the microarray experiment contributed with equal amounts of RNA to a pool. Sixty individuals were analyzed representing 6 biological replicates from 10 different experimental groups. Two separate control groups were used. Non-injected groups at the final temperatures (i.e. 10°C and 16°C) were used as prior to injection controls. At each time-point and temperatures there was also a sham-injected time matched control (injected with phosphate buffered saline [PBS]). The time points were 6 and 24 hours post-injection. Immunostimulated fish were injected with a weight-based dose of poly (I:C)(2 µg per g of wet body mass).

本实验旨在探究非致命性升温对大西洋鳕（Atlantic cod）抗病毒转录组应答的影响。针对经聚肌胞苷酸（poly(I:C)）注射并置于不同温度环境下的样本中筛选出的差异表达基因，本研究采用定量聚合酶链式反应（QPCR）进行了验证。本实验采用通用参考设计方案：微阵列实验中分析的每一份独立样本均按等量RNA比例混入同一RNA混合池。本次共分析60尾个体，涵盖10个不同实验组的6个生物学重复样本。实验共设置两类独立对照：分别以终温度为10℃与16℃的未注射对照组作为注射前对照；于各时间点与温度条件下，同时设置假注射对照（注射磷酸缓冲盐溶液（PBS）），且其采样时间与免疫刺激组完全匹配。实验采样时间点为注射后6小时与24小时。免疫刺激组鱼类按体重剂量注射聚肌胞苷酸，剂量为每克湿体重注射2微克。