Lacombe Research and Development Centre, Agriculture and Agri-Food Canada, Lacombe, Alberta, Canada.
Department of Animal Science, University of California Davis, Davis, California, United States of America.
PLoS One. 2021 May 25;16(5):e0251868. doi: 10.1371/journal.pone.0251868. eCollection 2021.
Evaluating RNA quality and transcriptomic profile of beef muscle over time post-mortem may provide insight into RNA degradation and underlying biological and functional mechanisms that accompany biochemical changes occurring post-mortem during transformation of muscle to meat. RNA was extracted from longissimus thoracis (LT) sampled from British Continental crossbred heifer carcasses (n = 7) stored at 4°C in an abattoir drip cooler at 5 time points post-mortem, i.e., 45 min (0 h), 6 h, 24 h, 48 h, and 72 h. Following RNA-Sequencing, processed reads were aligned to the ARS-UCD1.2 bovine genome assembly. Subsequent differential expression (DE) analysis identified from 51 to 1434 upregulated and 27 to 2256 downregulated DE genes at individual time points compared to time 0 h, showing a trend for increasing counts of both upregulated and downregulated genes over time. Gene ontology and biological pathway term enrichment analyses on sets of DE genes revealed several processes and their timelines of activation/deactivation that accompanied or were involved with muscle transformation to meat. Although the quality of RNA in refrigerated LT remained high for several days post-mortem, the expression levels of several known biomarker genes for meat quality began to change from 24 h onwards. Therefore, to ensure accuracy of predictions on meat quality traits based on the expression levels of those biomarker genes in refrigerated beef muscle tissue, it is crucial that those expression measurements be made on RNA sampled within 24 h post-mortem. The present study also highlighted the need for more research on the roles of mitochondrial genes and non-coding genes in orchestrating muscle tissue processes after death, and how pre-mortem immune status might influence post-mortem meat quality.
评估死后不同时间点牛肉肌肉的 RNA 质量和转录组谱,可能有助于深入了解 RNA 降解以及伴随死后生化变化发生的潜在生物学和功能机制,这些变化发生在肌肉向肉转化过程中。从在屠宰场滴冷器中以 4°C 储存的英国大陆杂交小母牛胴体(n = 7)的背最长肌(LT)中提取 RNA,在死后的 5 个时间点,即 45 分钟(0 小时)、6 小时、24 小时、48 小时和 72 小时时取样。在 RNA-Seq 之后,处理后的读数被映射到 ARS-UCD1.2 牛基因组组装上。随后的差异表达(DE)分析表明,与 0 小时相比,在单个时间点有 51 到 1434 个上调和 27 到 2256 个下调的 DE 基因,随着时间的推移,上调和下调基因的数量都呈现出增加的趋势。对 DE 基因集的基因本体和生物学途径术语富集分析揭示了几个过程及其激活/失活的时间线,这些过程伴随着或参与了肌肉向肉的转化。尽管冷藏 LT 的 RNA 质量在死后的几天内仍然很高,但一些已知的肉质生物标志物基因的表达水平从 24 小时开始发生变化。因此,为了确保基于冷藏牛肉肌肉组织中这些生物标志物基因的表达水平对肉质性状进行准确预测,至关重要的是,在死后 24 小时内对 RNA 进行采样。本研究还强调了需要更多研究线粒体基因和非编码基因在死后协调肌肉组织过程中的作用,以及生前免疫状态如何影响死后肉质。