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Identification of microRNAs controlling hepatic mRNA levels for metabolic genes during the metabolic transition from embryonic to posthatch development in the chicken

机译:在鸡从胚胎发育到孵化后的代谢过渡过程中,确定控制代谢基因肝mRNA水平的微小RNA

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Background The transition from embryonic to posthatch development in the chicken represents a massive metabolic switch from primarily lipolytic to primarily lipogenic metabolism. This metabolic switch is essential for the chick to successfully transition from the metabolism of stored egg yolk to the utilization of carbohydrate-based feed. However, regulation of this metabolic switch is not well understood. We hypothesized that microRNAs (miRNAs) play an important role in the metabolic switch that is essential to efficient growth of chickens. We used high-throughput RNA sequencing to characterize expression profiles of mRNA and miRNA in liver during late embryonic and early posthatch development of the chicken. This extensive data set was used to define the contributions of microRNAs to the metabolic switch during development that is critical to growth and nutrient utilization in chickens. Results We found that expression of over 800 mRNAs and 30 miRNAs was altered in the embryonic liver between embryonic day 18 and posthatch day 3, and many of these differentially expressed mRNAs and miRNAs are associated with metabolic processes. We confirmed the regulation of some of these mRNAs by miRNAs expressed in a reciprocal pattern using luciferase reporter assays. Finally, through the use of yeast one-hybrid screens, we identified several proteins that likely regulate expression of one of these important miRNAs. Conclusions Integration of the upstream regulatory mechanisms governing miRNA expression along with monitoring the downstream effects of this expression will ultimately allow for the construction of complete miRNA regulatory networks associated with the hepatic metabolic switch in chickens. Our findings support a key role for miRNAs in controlling the metabolic switch that occurs between embryonic and posthatch development in the chicken.
机译:背景技术鸡从胚胎发育到孵化后的过渡代表了从主要脂解代谢到主要脂生代谢的大规模代谢转换。这种代谢转换对于雏鸡成功地从储存的蛋黄的代谢转变为利用碳水化合物为基础的饲料至关重要。然而,对该代谢转换的调节尚不十分了解。我们假设microRNA(miRNA)在代谢转换中起重要作用,而代谢转换对于鸡的有效生长至关重要。我们使用高通量RNA测序来表征鸡胚胎后期和孵化后早期肝脏中mRNA和miRNA的表达谱。该广泛的数据集用于定义发育期间microRNA对代谢转换的贡献,这对鸡的生长和营养利用至关重要。结果我们发现,在胚胎第18天到孵化后第3天之间,胚胎肝中800多个mRNA和30个miRNA的表达发生了变化,并且这些差异表达的mRNA和miRNA中的许多与代谢过程有关。我们证实了使用萤光素酶报告基因检测法以互惠模式表达的miRNA对其中某些mRNA的调控。最后,通过使用酵母单杂交筛选,我们鉴定了可能调节这些重要miRNA之一表达的几种蛋白质。结论控制miRNA表达的上游调控机制的整合以及对该表达的下游效应的监测最终将允许构建与鸡肝代谢转换相关的完整miRNA调控网络。我们的发现支持miRNA在控制鸡胚胎和孵化后发育之间发生的代谢转换中的关键作用。

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