MicroRNA regulation below zero: Differential expression of miRNA-21 and miRNA-16 during freezing in wood frogs
MicroRNA regulation below zero: Differential expression of miRNA-21 and miRNA-16 during freezing in wood frogs
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DOI：
10.1016/j.cryobiol.2009.08.009
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发表时间：
2009-12-01
期刊：
影响因子：
2.7
通讯作者：
Storey, Kenneth
中科院分区：
文献类型：
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作者：
Biggar, Kyle K.;Dubuc, Adrian;Storey, Kenneth
自然的冷冻耐受性取决于许多生物化学适应，这些生物化学适应解决了冷冻对细胞施加的多重压力，并通过抑制冷冻状态下能量昂贵的细胞功能来保持活力。我们假设，microRNA，与mRNA结合的小的非编码RNA转录物，可以起到建立快速生物控制的作用，帮助重组冷冻生存的代谢优先级。使用RT-PCR在林蛙（Rana sylvatica）的肝脏和骨骼肌中评估选定的microRNA种类（miR-16和miR-21），比较对照（5 ℃驯化）与在-3 ℃冷冻24小时的动物。在冷冻过程中，肝脏和骨骼肌中miR-21的水平分别显著增加了1.5倍和1.3倍。在冷冻青蛙的肝脏中，miR-16的转录也显著增加了1.5倍，但在骨骼肌中下降了50%。Dicer（一种负责细胞质中成熟microRNA加工的III型RNA酶）的蛋白水平在肝脏中没有变化，在肌肉中显著降低了50%。该数据提供了第一份关于冷冻耐受脊椎动物中microRNA种类差异调节的报告，并提出了当动物过渡到冷冻状态时快速但可逆的基因沉默机制。皇冠版权所有（C）2009由爱思唯尔公司出版。All rights reserved.
Natural freeze tolerance depends on numerous biochemical adaptations that address the multiple stresses imposed on cells by freezing and preserves viability by suppressing energy-expensive cell functions in the frozen state. We hypothesized that microRNAs, small non-coding RNA transcripts that bind to mRNA, could act to establish rapid biological controls that aid the reorganization of metabolic priorities for freezing survival. Selected microRNA species (miR-16 and miR-21) were evaluated using RT-PCR in liver and skeletal muscle of wood frogs (Rana sylvatica) comparing controls (5 degrees C acclimated) with animals frozen for 24 h at -3 degrees C. Levels of miR-21 increased significantly during freezing by 1.5-fold and 1.3-fold in liver and skeletal muscle, respectively. MiR-16 transcripts also rose significantly by 1.5-fold in liver of frozen frogs but fell by 50% in skeletal muscle. Protein levels of Dicer, a type III RNase that is responsible for mature microRNA processing in the cytoplasm, were unchanged in liver and decreased significantly by 50% in muscle. This data provides the first report of differential regulation of microRNA species in a freeze tolerant vertebrate and suggest a mechanism for rapid, yet reversible, gene silencing when animals transition into the frozen state. Crown Copyright (C) 2009 Published by Elsevier Inc. All rights reserved.