Reductionist approaches in physiology and biochemistry are essential for understanding how animals cope and adapt to their environments. Transcriptomics is no longer restricted to a select few, and accessibility and affordability continue to facilitate its rapid growth as a science. More than 6000 publications (a conservative estimate) over the past decade quantify the response of the transcriptome to a wide breadth of questions in animal physiology. Transcriptomes have been quantified under conditions of hypoxia, climate change, salinity, drought, environmental pollution, and ultraviolet radiation among others; these studies have greatly improved understanding of the molecular machinery required for organismal adaptation. These “snapshots in time” however are never complete as the transcriptome is exquisitely sensitive to an individual's current physiologic state. Animal physiologists new to the field must recognize limitations of transcriptome technologies and consider experimental designs that strengthen physiologic interpretation. Current estimates suggest that a sample size of 6 or more are required for RNA-seq experiments in order to capture the majority of differentially expressed genes confidently. “Outside-the-box” approaches for statistical analyses of data derived from RNA-seq should be explored, as studies continue to point out that high false discoveries rates are pervasive with RNA-seq studies, reminiscent of the early days of microarrays. Incorporating biological variability, rather than reducing it (i.e. pooling strategies), into experimental designs is essential. Moreover, real-time PCR must not be viewed as a “validation step” to justify low samples sizes, but rather an orthogonal method to strengthen biological interpretation. The use of proper experimental controls in transcriptomics studies (i.e. spike-in controls and technical replication) are recommended and there is a pressing need for inter-laboratory tests (round robin experiments) to quantify repeatability and to identify sources of transcriptome variation within the context of animal physiology. Testing the reproducibility of transcriptome experiments in light of physiology in non-model organisms would be a significant contribution to the community. Single cell transcriptomics and multiplexing barcoding strategies such as decode-seq are poised to further advance the reductionist view of animal physiology; researchers are encouraged to consult literature herein and elsewhere for guidance on best practices and limitations of transcriptome technologies when studying the physiology of animals

还原论在生理学和生物化学中的方法对于理解动物如何应付和适应环境至关重要。转录组学不再仅限于少数几种，可及性和可负担性继续促进其作为一门科学的快速发展。在过去的十年中，超过6000种出版物（保守估计）量化了转录组对动物生理学广泛问题的响应。转录组已在低氧，气候变化，盐度，干旱，环境污染和紫外线辐射等条件下进行了定量；这些研究大大提高了对有机适应所需的分子机制的理解。但是，这些“及时的快照”从未完成，因为转录组对个人非常敏感。当前的生理状态。刚接触该领域的动物生理学家必须认识到转录组技术的局限性，并考虑加强生理学解释的实验设计。目前的估计表明，RNA-seq实验需要样本大小为6或更大，才能自信地捕获大多数差异表达的基因。随着研究不断指出，RNA-seq研究普遍存在高错误发现率，这令人回想起微阵列的早期发展，应该探索“开箱即用”的方法来对源自RNA-seq的数据进行统计分析。将生物变异性纳入实验设计中，而不是将其降低（即合并策略），这一点至关重要。此外，不得将实时PCR视为“验证步骤”以证明样本量小，而是加强生物学解释的正交方法。建议在转录组学研究中使用适当的实验对照（即加标对照和技术复制），并且迫切需要进行实验室间测试（循环实验）以量化可重复性并确定背景中转录组变异的来源动物生理学。根据生理学在非模式生物中测试转录组实验的可重复性，将对社区做出重大贡献。单细胞转录组学和诸如解码序列等多路复用条形码策略已准备就绪，以进一步推进动物生理学的简化论。