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生物钟基因周期在蟋蟀 Gryllus bimaculatus 的生物钟中起着重要作用。

The clock gene cycle plays an important role in the circadian clock of the cricket Gryllus bimaculatus.

机构信息

Graduate School of Natural Science and Technology, Okayama University, Okayama 700-8530, Japan.

出版信息

J Insect Physiol. 2013 Jul;59(7):697-704. doi: 10.1016/j.jinsphys.2013.04.011. Epub 2013 May 7.

Abstract

To dissect the molecular oscillatory mechanism of the circadian clock in the cricket Gryllus bimaculatus, we have cloned a cDNA of the clock gene cycle (Gb'cyc) and analyzed its structure and function. Gb'cyc contains four functional domains, i.e. bHLH, PAS-A, PAS-B and BCTR domains, and is expressed rhythmically in light dark cycles, peaking at mid night. The RNA interference (RNAi) of Clock (Gb'Clk) and period (Gb'per) reduced the Gb'cyc mRNA levels and abolished the rhythmic expression, suggesting that the rhythmic expression of Gb'cyc is regulated by a mechanism including Gb'Clk and Gb'per. These features are more similar to those of mammalian orthologue of cyc (Bmal1) than those of Drosophila cyc. A single treatment with double-stranded RNA (dsRNA) of Gb'cyc effectively knocked down the Gb'cyc mRNA level and abolished its rhythmic expression. The cyc RNAi failed to disrupt the locomotor rhythm, but lengthened its free-running period in constant darkness (DD). It is thus likely that Gb'cyc is involved in the circadian clock machinery of the cricket. The cyc RNAi crickets showed a rhythmic expression of Gb'per and timeless (Gb'tim) in the optic lobe in DD, explaining the persistence of the locomotor rhythm. Surprisingly, cyc RNAi revealed a rhythmic expression of Gb'Clk in DD which is otherwise rather constitutively expressed in the optic lobe. These facts suggest that the cricket might have a unique clock oscillatory mechanism in which both Gb'cyc and Gb'Clk are rhythmically controlled and that under abundant expression of Gb'cyc the rhythmic expression of Gb'Clk may be concealed.

摘要

为了剖析蟋蟀(Gryllus bimaculatus)生物钟的分子振荡机制,我们克隆了生物钟基因周期(Gb'cyc)的 cDNA,并分析了其结构和功能。Gb'cyc 含有四个功能域,即 bHLH、PAS-A、PAS-B 和 BCTR 结构域,并呈现出昼夜节律性表达,午夜达到峰值。对 Clock(Gb'Clk)和 period(Gb'per)的 RNA 干扰(RNAi)降低了 Gb'cyc mRNA 水平并消除了其节律性表达,表明 Gb'cyc 的节律性表达受到包括 Gb'Clk 和 Gb'per 的机制调节。这些特征与哺乳动物生物钟基因(Bmal1)的特征更为相似,而与果蝇的 cyc 基因(Drosophila cyc)的特征不同。单独用双链 RNA(dsRNA)处理 Gb'cyc 可有效降低 Gb'cyc mRNA 水平并消除其节律性表达。cyc RNAi 未能破坏运动节律,但延长了其在连续黑暗(DD)中的自由运行周期。因此,Gb'cyc 可能参与了蟋蟀的生物钟机制。在 DD 中,cyc RNAi 蟋蟀的 Gb'per 和 timeless(Gb'tim)在视神经叶中表现出节律性表达,解释了运动节律的持续。令人惊讶的是,cyc RNAi 在 DD 中揭示了 Gb'Clk 的节律性表达,而 Gb'Clk 在视神经叶中通常是组成性表达的。这些事实表明,蟋蟀可能具有独特的时钟振荡机制,其中 Gb'cyc 和 Gb'Clk 均受到节律性控制,并且在 Gb'cyc 大量表达的情况下,Gb'Clk 的节律性表达可能被掩盖。

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