RNA interference of timeless gene does not disrupt circadian locomotor rhythms in the cricket Gryllus bimaculatus
RNA interference of timeless gene does not disrupt circadian locomotor rhythms in the cricket Gryllus bimaculatus
复制标题
DOI：
10.1016/j.jinsphys.2010.07.002
复制
发表时间：
2010-12-01
影响因子：
2.2
通讯作者：
Tomioka, Kenji
中科院分区：
文献类型：
--
作者：
Danbara, Yoshiki;Sakamoto, Tomoaki;Tomioka, Kenji
分子研究表明，由所谓的时钟基因组成的自动调节负反馈环路构成了果蝇的生物钟。然而，这一假设在其他昆虫中并未得到充分支持，因此有待检验。在蟋蟀双斑蟋蟀中，我们之前已经表明周期（per）在节律产生中起着至关重要的作用。在本研究中，我们克隆了时钟基因 timeless（rim）的 cDNA，并使用RNA 干扰 蟋蟀 tim 的分子结构与其他昆虫物种具有相当高的相似性 实时 RT-PCR 分析显示 rim mRNA 在 LD 和 DD 中均表现出节律性表达，与（主观）夜间的每个峰值相似 当注射 rim 双链 RNA (dstim) tim mRNA 水平显着降低，其昼夜节律表达节律被消除 dstim 处理后，但成年蟋蟀在 DD 中表现出明显的运动节律，自由奔跑期明显短于注射 Discosoma sp Red2 (DsRed2) dsRNA 的对照蟋蟀 这些结果表明，蟋蟀边缘在自由奔跑期的微调中发挥着一定作用，但对于生物钟的振荡可能并不重要 (C) 2010 Elsevier Ltd 版权所有
Molecular studies revealed that autoregulatory negative feedback loops consisting of so-called clock genes constitute the circadian clock in Drosophila However this hypothesis is not fully supported in other Insects and is thus to be examined In the cricket Gryllus bimaculatus we have previously shown that period (per) plays an essential role in the rhythm generation In the present study we cloned cDNA of the clock gene timeless (rim) and investigated its role in the cricket circadian oscillatory mechanism using RNA interference Molecular structure of the cricket tim has rather high similarity to those of other insect species Real-time RT-PCR analysis revealed that rim mRNA showed rhythmic expression in both LD and DD similar to that of per peaking during the (subjective) night When injected with rim double-stranded RNA (dstim) tim mRNA levels were significantly reduced and its circadian expression rhythm was eliminated After the dstim treatment however adult crickets showed a clear locomotor rhythm in DD with a free-running period significantly shorter than that of control crickets injected with Discosoma sp Red2 (DsRed2) dsRNA These results suggest that in the cricket rim plays some role in fine-tuning of the free-running period but may not be essential for oscillation of the circadian clock (C) 2010 Elsevier Ltd All rights reserved