光和投喂时间对大海马生长及生物钟基因和脂类代谢基因表达的影响

LIGHT AND FEEDING TIME ON THE GROWTH, BIOLOGICAL CLOCK GENE EXPRESSION, AND LIPID METABOLISM GENE EXPRESSION IN HIPPOCAMPUS KUDA BLEEKER

  • 摘要: 生物节律在生物的代谢活动中扮演非常重要的角色。文章以大海马(Hippocampus kuda)为研究对象, 分别设置2个光周期(L﹕D=16﹕8、L﹕D=12﹕12)和2个投饵时间策略(开灯后2h、关灯前2h), 分析大海马的生物钟系统之间以及生物钟与脂类代谢之间的关系, 探究大海马人工养殖的适宜环境条件以及其体内重要脂类物质的合成代谢条件。结果表明: 在所有组别中, 检测的7个钟基因中有6个基因(Clock、Bmal1、Per1、Per2、Per3Cry1)在大海马的脑和肝脏中都有明显的节律性表达, 说明生物钟基因表达与光有较密切的关系。其中, 大海马在长光亮期(16:8)及开灯后2h投喂的条件下, 其脑中的中枢钟基因与肝脏中的外周钟基因间的协同性最佳, 表明该条件对大海马生长较为有利。此外, 大海马肝脏中8个脂类代谢相关基因(Hmgcr、Mvk、Mvd、Lss、Fdps、Cetp、Scap、Srebp1)在开灯后2h投喂的2个组别中都呈现了节律性表达, 而在关灯前2h投喂的2个组别中仅Hmgcr、Mvk、Scap、Srebp1等4个基因(L﹕D=16﹕8)和Hmgcr、Mvk、Scap、Mvd、Srebp1等5个基因(L﹕D=12﹕12)有明显节律性表达, 说明脂类代谢相关基因的表达与光影响并不非常密切, 大部分基因的表达受到了投喂时间的影响。总之, 实验证实了大海马中存在有完整的生物钟系统, 该生物钟系统与脂类代谢间存在紧密的联系。通过光和投喂时间的综合分析, 初步推断大海马养殖的适宜条件为长光亮周期及在有光时及时给饵(开灯后2h内), 该条件下能促使大海马的钟基因与代谢基因都呈现稳定的节律性表达, 且中枢钟系统与外周钟系统间的协同性达到最佳。同时, 也能较好地促进脂类物质的代谢合成和积累。

     

    Abstract: Biological rhythms play a critical role in the metabolism of organisms. In this study, the juvenile Hippocampus kuda was used as the research object. Four treatments were established, including two photoperiods (L (light)﹕D (dark) 16﹕8, 12﹕12) and two feeding times (2h after light-on the and 2h before light-off), to analyze the interaction between the two biological clock systems of H. kuda and their relationship with nutrient metabolism. The aim was to determine the optimal environmental conditions for artificial breeding and the conditions necessary for the synthesis of important metabolites in H. kuda. Results showed that under long photoperiod (L﹕D16﹕8) with feeding 2h after light-on, H. kuda exhibited the highest specific growth rate in body length (SGRL) and body weight (SGRW), as well as the highest condition factor (CF) in all four treatments. Rhythmic expressions were observed in six clock genes (Clock, Bmal1, Per1, Per2, Per3, and Cry1) and eight lipid-related metabolic genes (Hmgcr, Mvk, Mvd, Lss, Fdps, Cetp, Scap, and Srebp1). The trend of clock gene expression in the brain throughout the day, and its peak expression, was consistent with those in liver, which correlated with SGRL, SGRW, and CF. However, in the treatments where feeding occurred 2h before light-off, these parameters were significantly lower than those with feeding 2h after light-on. Additionally, the central clock system and the trend of clock gene expression in the brain throughout the day, including peak expression times, differed completely from those in the liver. Only four lipid-related metabolic genes (Hmgcr, Mvk, Scap, and Srebp1) exhibited rhythmic expression under the long photoperiod, while five lipid-related metabolic genes (Hmgcr, Mvk, Mvd, Scap, and Srebp1) exhibited rhythmic expressions under the short photoperiod. The study suggests that H. kuda possesses a comprehensive biological clock system closely linked to lipid metabolism. Combined analysis indicated that stable rhythmic expression of clock genes and metabolic genes could be maintained under the conditions of L:D=16:8 with feeding 2hours after light-on. Under these conditions, the clock gene expression in the brain and liver tended to synchronize, promoting the growth of H. kuda and maintaining the normal lipid metabolism pathway. These findings provide a scientific basis for promoting lipid synthesis and accumulation in H. kuda.

     

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