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光和摄食对海洋硬骨鱼(真鲷)分子生物钟的驯化作用。

Light and feeding entrainment of the molecular circadian clock in a marine teleost (Sparus aurata).

机构信息

Department of Physiology, Faculty of Biology, Regional Campus of International Excellence Campus Mare Nostrum, University of Murcia, Murcia, Spain.

出版信息

Chronobiol Int. 2013 Jun;30(5):649-61. doi: 10.3109/07420528.2013.775143. Epub 2013 May 20.

Abstract

Daily light and feeding cycles act as powerful synchronizers of circadian rhythmicity. Ultimately, these external cues entrain the expression of clock genes, which generate daily rhythmic behavioral and physiological responses in vertebrates. In the present study, we investigated clock genes in a marine teleost (gilthead sea bream). Partial cDNA sequences of key elements from both positive (Bmal1, Clock) and negative (Per2, Cry1) regulatory loops were cloned before studying how feeding time affects the daily rhythms of locomotor activity and clock gene expression in the central (brain) and peripheral (liver) oscillators. To this end, all fish were kept under a light-dark (LD) cycle and were divided into three experimental groups, depending on the time of their daily meal: mid-light (ML), mid-darkness (MD), or at random (RD) times. Finally, the existence of circadian control on gene expression was investigated in the absence of external cues (DD + RD). The behavioral results showed that seabream fed at ML or RD displayed a diurnal activity pattern (>91% of activity during the day), whereas fish fed at MD were nocturnal (89% of activity during the night). Moreover, seabream subjected to regular feeding cycles (ML and MD groups) showed food-anticipatory activity (FAA). Regardless of the mealtime, the daily rhythm of clock gene expression in the brain peaked close to the light-dark transition in the case of Bmal1 and Clock, and at the beginning of the light phase in the case of Per2 and Cry1, showing the existence of phase delay between the positive and negative elements of the molecular clock. In the liver, however, the acrophases of the daily rhythms differed depending on the feeding regime: the maximum expression of Bmal1 and Clock in the ML and RD groups was in antiphase to the expression pattern observed in the fish fed at MD. Under constant conditions (DD + RD), Per2 and Cry1 showed circadian rhythmicity in the brain, whereas Bmal1, Clock, and Per2 did in the liver. Our results indicate that the seabream clock gene expression is endogenously controlled and in liver it is strongly entrained by food signals, rather than by the LD cycle, and that scheduled feeding can shift the phase of the daily rhythm of clock gene expression in a peripheral organ (liver) without changing the phase of these rhythms in a central oscillator (brain), suggesting uncoupling of the light-entrainable oscillator (LEO) from the food-entrainable oscillator (FEO). These findings provide the basis and new tools for improving our knowledge of the circadian system and entraining pathways of this fish species, which is of great interest for the Mediterranean aquaculture.

摘要

日常的光照和摄食周期是调节昼夜节律的强大同步因子。最终,这些外部线索使时钟基因的表达同步,从而在脊椎动物中产生每日的节律性行为和生理反应。在本研究中,我们研究了一种海洋硬骨鱼(金头鲷)中的时钟基因。在研究摄食时间如何影响中央(脑)和外周(肝)振荡器的运动活性和时钟基因表达的昼夜节律之前,我们克隆了正调控(Bmal1、Clock)和负调控(Per2、Cry1)环路的关键元件的部分 cDNA 序列。为此,所有的鱼都在光照-黑暗(LD)循环下饲养,并根据每天的摄食时间分为三组实验:光照中期(ML)、黑暗中期(MD)或随机时间(RD)。最后,在没有外部线索的情况下(DD+RD),研究了基因表达的昼夜节律控制。行为学结果表明,ML 或 RD 摄食的金头鲷表现出昼间活动模式(白天活动超过 91%),而 MD 摄食的鱼则表现为夜间活动(夜间活动超过 89%)。此外,在定期摄食循环(ML 和 MD 组)下,金头鲷表现出摄食前活动(FAA)。无论摄食时间如何,脑时钟基因表达的昼夜节律在 Bmal1 和 Clock 时接近明暗转换,在 Per2 和 Cry1 时接近光相开始,表明分子钟的正负元件之间存在相位延迟。然而,在肝脏中,摄食模式的不同导致每日节律的高峰出现在不同的时间:ML 和 RD 组的 Bmal1 和 Clock 的最大表达与 MD 摄食鱼的表达模式相反。在恒常条件下(DD+RD),脑内的 Per2 和 Cry1 表现出昼夜节律性,而肝脏中的 Bmal1、Clock 和 Per2 则表现出昼夜节律性。我们的结果表明,金头鲷的时钟基因表达是内源性控制的,在肝脏中,它强烈地受到食物信号的同步,而不是 LD 循环的同步,定时摄食可以改变外周器官(肝脏)中时钟基因表达的昼夜节律的相位,而不会改变中央振荡器(大脑)中这些节律的相位,表明光可诱导振荡器(LEO)与食物可诱导振荡器(FEO)的解耦。这些发现为我们了解这种鱼类的昼夜节律系统和同步途径提供了基础和新工具,这对地中海水产养殖具有重要意义。

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