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RNAseq Analysis of Rodent Spaceflight Experiments Is Confounded by Sample Collection Techniques

机译:啮齿动物空间实验的RNASEQ分析由样品收集技术混淆

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摘要

To understand the physiological changes that occur in response to spaceflight, mice are transported to the International Space Station (ISS) and housed for variable periods of time before euthanasia on-orbit or return to Earth. Sample collection under such difficult conditions introduces confounding factors that need to be identified and addressed. We found large changes in the transcriptome of mouse tissues dissected and preserved on-orbit compared with tissues from mice euthanized on-orbit, preserved, and dissected after return to Earth. Changes due to preservation method eclipsed those between flight and ground samples, making it difficult to identify spaceflight-specific changes. Follow-on experiments to interrogate the roles of euthanasia methods, tissue and carcass preservation protocols, and library preparation methods suggested that differences due to preservation protocols are exacerbated when coupled with polyA selection. This has important implications for the interpretation of existing datasets and the design of future experiments.
机译:要了解响应太空飞行的生理变化,小鼠将被运送到国际空间站(ISS),并在安乐死的轨道前或返回地球前进行可变时间段。在这种困难条件下的样品收集介绍了需要识别和解决的混淆因素。我们发现与来自小鼠的组织的小鼠组织的转录组中的转录组大变化与来自小鼠的组织安乐死,保存在滚动后,并在返回地球后解剖。由于保存方法因飞行和地面样本而被蚀的变化,使得难以识别特定的空间变化。后续实验以询问安乐死方法,组织和胴体保存方案的作用,文库制备方法表明,当与PolyA选择结合时,由于保存方案引起的差异而加剧。这对对现有数据集的解释和未来实验的设计具有重要意义。

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