【作者】 吴萍；

【导师】 郭新红； 张建设；

【作者基本信息】 湖南大学 ， 生物医学工程， 2016， 硕士

【摘要】 生物钟是指机体内节律性的活动,能使机体维持持续而恒定的规律变化,即使外界条件变化也能保持恒定。机体的主要控制生物节律的器官在下丘脑,但外周组织也具有昼夜节律,例如肌肉。鱼肌纤维类型与人体相似,分为主要负责代谢的快肌和主要进行有氧运动的慢肌,哺乳动物的两种肌纤维是嵌合式,白肌纤维和红肌纤维交叉散漫分布,难分离,而鱼类的快慢肌很有特色,易完全分离。目前对于肌肉生物钟的研究也比较少。本论文对从鳜红肌筛选到的15个时钟基因(arntl1、arntl2、clock、cry1、cry2、cry3、cry-dash、 npas2、nr1d1、nr1d2、per1、 per2、per3、Rora和tim),将所有时钟基因进行系统进化树发现,在硬骨鱼类的时钟基因与其他鱼类的亲缘性较近,序列相对保守,也证明了转录组库内时钟基因序列及功能预测的准确性。快肌中,时钟基因arntl1、arntl2、clock、cry1、nr1d1、 nr1d2、per1、per2、per3和Rora,功能基因mbnl1、mrf4、mstn、myoD、myoG、 myf5和Jpena有昼夜节律性。节律性时钟基因之间有正相关性,也有负相关性。功能基因pena与arntl2, cry1和per1负相关。mbnl1与arntl2、cry1、nr1d1、per2和per3有强的正关联。myoG也与arntl2和per2强正相关。在慢肌中,时钟基因arntl1、 arntl2、clock、cry1、cry3、npas2、nr1d1、per1、per2、Rora和tim,功能基因foxk2、 mrf4、mstn、myf5和pena有昼夜节律性。节律性的时钟基因之间只有正关联,并没有负关联。pena与arntl2, cry1和per1呈中度正相关。myf5, mrf4, pcna和mstn在慢肌表现出与cry3和npas2很强的相关性。肌肉功能基因foxk2也显示出与正调控因子arntl2有很强的相关性。结果显示不管是在快肌还是慢肌中,多基因之间都有强关联性,这些时钟基因与肌肉的功能基因互相协调和拮抗其功能,共同对鳜的肌纤维生长代谢和生物钟调控,这也对于进一步研究他们之间的作用关系提供了依据。更多还原

【Abstract】 Biological clock is the activity of the body’s internal rhythm that can make the organism to maintain a constant change, even if the external conditions change. The organism biological rhythm is mainly controlled by the hypothalamus, but the peripheral tissues also have circadian rhythms, such as muscle. Fish muscle fiber types are similar to those of human that contain the fast muscle and slow muscle which was mainly responsible for the metabolism and maintained oxygen movement, respectively. In fish fast and slow muscle is quite distinctive and easy separation while in mammals it is embedded and distribution which scattered and difficult to separate. At present, there are few studies on muscle biological clock. This paper screened 15 clock genes (arntl1、arntl2、 clock、cry1、cry2、cry3、cry-dash、npas2、nr1d1、nr1d2、per1、per2、per3、 Rora and tim) from Chinese Perch red muscle transcriptome, and all clock gene made phylogenetic tree found that they were closely related to their orthologs in other teleost fish. These results proved transcriptome library clock gene sequence and function prediction accuracy. In the fast muscle, the clock genes arntll, arntl2, clock, cry1, nr1d1, nr1d2, per1, per2, per3 and Rora, functional genes mbnll, mrf4, mstn, myoD, myoG, myf5 and pcna had daily rhythmicity. There is a positive and negative correlation between the rhythms of clock genes. The pcna showed moderate negative correlation with arntl2, cry1 and per1. The mbnll positively correlated strong with the components of the the daily expression of arntl1, cry1, nr1d1, per1, per3 and clock. The myoG also strong positively correlated with arntl1 and per1. In the slow muscle, the clock gene arntll, arntl2, clock, cry1, cry3, npas2, nr1d1, per1, per2. Rora and tim, functional genes including foxk2, mrf4, mstn, myf5 and pcna displayed daily rhythmic expression. Rhythm of clock genes only had positive correlation. Transcript levels of pcna positively correlated moderate with the components of arntl2, cry1 and per1. The myf5, mrf4, pcna and mstn in the slow muscle showed strong correlation with cry3 and npas2. The functional gene foxk2 also showed strong correlation with the components of the transcriptional activator arm arntl2. The results showed that, regardless of the fast or slow muscle, there was a strong correlation between multiple genes. These clock and functional genes firstly co-ordinate and antagonize its function, then regulate Chinese Perch muscle fiber growth, metabolism and the biological clock. The above results also provided the basis for further study of the relationship between them.更多还原