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大鼠视网膜中的生物钟基因表达:光照条件和光感受器退化的影响。

Clock gene expression in the rat retina: effects of lighting conditions and photoreceptor degeneration.

作者信息

Tosini Gianluca, Kasamatsu Manami, Sakamoto Katsuhiko

机构信息

Circadian Rhythms and Sleep Disorders Program, Neuroscience Institute, Morehouse School of Medicine, 720 Westview Dr, Atlanta, GA 30310-1495, USA.

出版信息

Brain Res. 2007 Jul 23;1159:134-40. doi: 10.1016/j.brainres.2007.05.023. Epub 2007 May 23.

Abstract

Previous studies have shown that, in the Royal College of Surgeon rat, circadian rhythms in the retinal dopaminergic and melatonergic systems are still present after the photoreceptors have degenerated, thus demonstrating that circadian rhythmicity in the mammalian retina can be generated independently from the photoreceptors. The aim of the present study was to investigate the pattern of expression of the clock genes in the retina of the Royal College of Surgeons rat under different lighting conditions. Expression of clock genes was investigated in the retina of normal and dystrophic Royal College of Surgeons rats under 12 h of light/12 h of dark (LD), constant darkness (DD) and constant light (LL) using Real Time Quantitative RT-PCR. Our data indicate that, in control animals, Period1, Period2, Cryptochrome1, Cryptochrome2, Clock, Rora, Rev-Erb alpha and Npas2 mRNA levels showed a significant variation over the sampling period in LD cycles and in DD, whereas Bmal1 mRNA did not show any significant variation. In LL, the transcripts for Per1, Per2, Clock and Rev-Erb alpha showed significant temporal variations. In the dystrophic retina, only Per1 and Per2 mRNA levels showed a temporal variation over the 20-h period. Our work indicates that degeneration of the photoreceptor cells dramatically affected the expression levels and patterns of many clock genes. Finally, the present study suggests that investigating the expression pattern of clock genes using the whole retina or animals with photoreceptor degeneration may not provide any definitive answers about the working of the retinal circadian clock system.

摘要

先前的研究表明,在皇家外科学院大鼠中,视网膜多巴胺能和褪黑素能系统的昼夜节律在光感受器退化后仍然存在,从而证明哺乳动物视网膜中的昼夜节律性可以独立于光感受器产生。本研究的目的是调查不同光照条件下皇家外科学院大鼠视网膜中生物钟基因的表达模式。使用实时定量逆转录聚合酶链反应(Real Time Quantitative RT-PCR),研究正常和营养不良的皇家外科学院大鼠在12小时光照/12小时黑暗(LD)、持续黑暗(DD)和持续光照(LL)条件下视网膜中生物钟基因的表达。我们的数据表明,在对照动物中,周期蛋白1(Period1)、周期蛋白2(Period2)、隐花色素1(Cryptochrome1)、隐花色素2(Cryptochrome2)、生物钟(Clock)、视黄酸相关孤儿受体α(Rora)、视黄酸受体相关孤儿受体α(Rev-Erb alpha)和神经元 PAS 结构域蛋白 2(Npas2)的 mRNA 水平在 LD 周期和 DD 的采样期间显示出显著变化,而脑和肌肉 ARNT 样蛋白 1(Bmal1)的 mRNA 没有显示出任何显著变化。在 LL 条件下,Per1、Per2、Clock 和 Rev-Erb alpha 的转录本显示出显著的时间变化。在营养不良的视网膜中,只有 Per1 和 Per2 的 mRNA 水平在20小时期间显示出时间变化。我们的工作表明,光感受器细胞的退化显著影响了许多生物钟基因的表达水平和模式。最后,本研究表明,使用整个视网膜或光感受器退化的动物来研究生物钟基因的表达模式可能无法为视网膜昼夜节律时钟系统的工作提供任何明确的答案。

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