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花鲈垂体和下丘脑中生物钟基因在3种光周期下的表达节律分析

袁满 王鹏飞 闫路路 赵超 范嗣刚 陈祥 邱丽华

袁满, 王鹏飞, 闫路路, 赵超, 范嗣刚, 陈祥, 邱丽华. 花鲈垂体和下丘脑中生物钟基因在3种光周期下的表达节律分析[J]. 南方水产科学. doi: 10.12131/20200113
引用本文: 袁满, 王鹏飞, 闫路路, 赵超, 范嗣刚, 陈祥, 邱丽华. 花鲈垂体和下丘脑中生物钟基因在3种光周期下的表达节律分析[J]. 南方水产科学. doi: 10.12131/20200113
Man YUAN, Pengfei WANG, Lulu YAN, Chao ZHAO, Sigang FAN, Xiang CHEN, Lihua QIU. Circadian rhythmicity of clock genes in pituitary and hypothalamus of spotted sea perch (Lateolabrax maculates) under three photoperiod conditions[J]. South China Fisheries Science. doi: 10.12131/20200113
Citation: Man YUAN, Pengfei WANG, Lulu YAN, Chao ZHAO, Sigang FAN, Xiang CHEN, Lihua QIU. Circadian rhythmicity of clock genes in pituitary and hypothalamus of spotted sea perch (Lateolabrax maculates) under three photoperiod conditions[J]. South China Fisheries Science. doi: 10.12131/20200113

花鲈垂体和下丘脑中生物钟基因在3种光周期下的表达节律分析

doi: 10.12131/20200113
基金项目: 国家自然科学基金项目 (31802281);广东省自然科学基金项目 (2018A030313137);广州市珠江科技新星项目 (201906010028);中国水产科学研究院基本科研业务费专项 (2020XT09,2020TD21)
详细信息
    作者简介:

    袁满:袁 满 (1996—),女,硕士研究生,研究方向为鱼类繁殖生物学。E-mail: madisony@163.com

    通讯作者:

    邱丽华 (1971—),女,博士,研究员,从事水产生物技术研究。E-mail: qiugroup_bio@outlook.com

  • 中图分类号: S 917.4

Circadian rhythmicity of clock genes in pituitary and hypothalamus of spotted sea perch (Lateolabrax maculates) under three photoperiod conditions

  • 摘要: 花鲈 (Lateolabrax maculatus) 是中国重要的水产养殖鱼类,其繁殖活动受光周期调控。文章研究了3种光周期 [ 光 (L) 暗 (D) 比分别为16L∶8D、12L∶12D和8L∶16D] 条件下,7个重要生物钟基因 (Bmal2、Npas4、Per2、Cry1、Cry1aCry2 及Timeless) 在花鲈垂体和下丘脑中的昼夜表达规律。结果表明,在12L∶12D条件下垂体中Per2、Cry1、Cry2、Cry1aTimeless表现出昼夜节律性,下丘脑中Per2、Cry2、Cry1、Timeless表现出昼夜节律性,相同基因在垂体和下丘脑两种组织中的昼夜节律不同,长光照或短光照会改变昼夜节律的震荡强弱,也会改变峰值相位,部分基因在长光照或短光照下会出现失去昼夜节律性的现象。
  • 图  1  3种光周期下花鲈垂体中生物钟基因mRNA昼夜节律表达的时间模式

    每个点的值代表每个生物钟基因的表达水平,采用“平均值±标准差”表示 (n=3);不同字母表示显著性差异 (P<0.05);虚线表示由余弦分析计算出的基因昼夜节律表达余弦函数,图上方条状图表示光周期,白色代表光照,黑色代表黑暗;后图同此

    Figure  1.  Relative expression of clock genes in pituitary of L. maculatus under three photoperiod conditions

    The values are $ \overline X \pm {\rm{SD}} $ (n=3) of the normalized transcript levels of each clock gene. Different lowercase letters indicate significant difference (P<0.05). The broken line is the periodic sinusoidal function of gene expression in a circadian cycle constructed from the periodicity parameters. The photoperiod regime is represented by composite block above the graph. White and black represent the light and dark phase, respectively; the same case in the following figure.

    图  2  3种光周期下花鲈下丘脑中生物钟基因mRNA昼夜节律表达的时间模式

    Figure  2.  Relative expression of clock genes in hypothalamus of L. maculatus under three photoperiod conditions

    表  1  荧光定量引物

    Table  1.   Primers for qRT-PCR

    引物名称
    Primer name
    序列 (5'–3')
    Sequence
    qBmal2-F TCTGAAAGTACAGGCGAGCCGTCCCA
    qBmal2-R CAGTGTAAGTCATCAAAGTCCCCAGT
    qPer2-F CCCCACCGTCCTTCAG
    qPer2-R TCCCATTCAGCCGCATTA
    qNpas4-F GTCATCTCCTGTGTCCTCTTGCT
    qNpas4-R ACTTCCACTCCCATCTTTGTG
    qCry1-F GACTGGGCTCTGAATGCTGGAA
    qCry1-R TGCCTGCTGAATACTGCGTGGAG
    qCry1a-F CAAAGCAGTATGGGCAGGT
    qCry1a-R AGTAGAAGAGCCGACAGGAGA
    qCry2-F GTCAATGCTGGCAGTTGGATGTGG
    qCry2-R GGGATGTAACGCCTGATGTATTCT
    qTimeless-F GAAACCAGACAGCCTCACTCCTAC
    qTimeless-R AAAGACTCCGACAACTGAAACCCT
    qβ-actin-F CAACTGGGATGACATGGAGAAG
    qβ-actin-R TTGGCTTTGGGGTTCAGG
    下载: 导出CSV

    表  2  花鲈垂体中生物钟基因mRNA表达的昼夜节律性参数

    Table  2.   Circadian rhythmic parameters of clock genes mRNA expressions in pituitary of L. maculatus

    基因
    Gene
    光周期
    Photoperiod
    振幅
    Amplitude
    峰值相位
    Acrophase
    中值
    Mesor
    P
    Bmal2 16L∶8D 0.471
    12L∶12D 0.416
    8L∶16D 0.200
    Naps4 16L∶8D 0.070
    12L∶12D 0.274
    8L∶16D 0.286
    Per2 16L∶8D 41.084 1 −18.890 5 43.894 7 <0.001
    12L∶12D 0.253 6 0.226 6 0.187 7 <0.001
    8L∶16D 0.371 2 −0.366 8 0.452 4 0.001
    Cry1 16L∶8D 0.951 9 9.299 7 1.390 5 <0.001
    12L∶12D 1.314 0 0.420 3 1.455 6 <0.001
    8L∶16D 1.165 7 10.981 0 1.647 0 <0.001
    Cryla 16L∶8D 0.364 2 6.375 6 1.577 3 0.002
    12L∶12D 0.498 9 0.568 9 0.648 5 <0.001
    8L∶16D 0.238 8 −7.008 3 1.027 7 <0.001
    Cry2 16L∶8D 47.911 9 0.018 2 31.143 5 <0.001
    12L∶12D 3.106 3 15.143 0 2.599 7 <0.001
    8L∶16D 29.750 8 −16.533 1 34.491 7 <0.001
    Timeless 16L∶8D 0.354 5 −4.266 8 0.860 5 <0.001
    12L∶12D 3.368 8 −1.157 9 2.785 8 <0.001
    8L∶16D 1.264 4 10.797 5 2.447 2 <0.001
    注:振幅为拟合波形峰值之间距离的一半;中值为周期平均值;峰值为相位周期最高幅度的时间点 (弧度);P为时间点间的差异;后表同此 Note: The amplitude is half of the distance between the peak values of the fitting waveform; the median value is the periodic average value; the peak value is the time point (radian) with the highest amplitude of phase period; P is the difference between time points; the same case in the following table.
    下载: 导出CSV

    表  3  花鲈下丘脑中生物钟基因mRNA表达的昼夜节律性参数

    Table  3.   Circadian rhythmic parameters of clock genes mRNA expressions in hypothalamus of L. maculatus

    基因
    Gene
    处理组
    Photoperiod
    振幅
    Amplitude
    峰值相位
    Acrophase
    中值
    Mesor
    P
    Bmal2 16L∶8D 0.469
    12L∶12D 0.060
    8L∶16D 0.260
    Naps4 16L∶8D 0.762
    12L∶12D 0.166
    8L∶16D 0.095
    Per2 16L∶8D 0.472 1 −8.423 1 0.808 7 0.018
    12L∶12D 2.773 3 −8.478 7 3.971 5 0.001
    8L∶16D 0.896 2 −11.462 8 1.088 0 0.004
    Cry1 16L∶8D 0.436 6 −5.273 3 1.408 4 0.006
    12L∶12D 0.378 3 0.461 1 1.654 0 0.007
    8L∶16D 1.741 5 −0.099 9 1.554 6 <0.001
    Cryla 16L∶8D 0.096 4 −1.274 2 0.876 5 0.047
    12L∶12D 1.000 0 1.436 0 0.000 3 0.017
    8L∶16D 1.038 8 −9.228 1 1.978 6 <0.001
    Cry2 16L∶8D 44.338 7 7.936 4 24.514 2 <0.001
    12L∶12D 2.681 0 8.881 4 4.563 0 <0.001
    8L∶16D 6.819 8 8.026 9 4.909 8 <0.001
    Timeless 16L∶8D 9.888 2 −4.951 6.863 4 <0.001
    12L∶12D 2.955 7 9.236 6 3.071 5 <0.001
    8L∶16D 78.572 9 −6.378 3 45.208 1 <0.001
    下载: 导出CSV
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  • 收稿日期:  2020-05-22
  • 修回日期:  2020-07-21
  • 网络出版日期:  2020-11-19

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