17α-甲基睾酮对稀有鮈鲫精巢DNA甲基转移酶基因表达的影响

doi:10.12160/j.issn.1672-5190.2022.02.001

摘要/Abstract

摘要： [目的] 以稀有鮈鲫雄鱼为研究对象,探讨17α-甲基睾酮（17α-methyltestosterone,MT）对稀有鮈鲫精巢甲基转移酶（DNMT）基因相对表达量的影响。[方法] 采用不同浓度MT（25、50和100 ng/L）处理稀有鮈鲫雄鱼7、14和21 d,处理结束时,每组随机解剖10条鱼,取精巢组织,Trizol一步法提取总RNA,实时荧光定量PCR（qRT-PCR）检测DNA甲基转移酶基因（dnmts）相对表达量。[结果] MT处理稀有鮈鲫雄鱼7 d,25 ng/L MT处理组（P<0.05）、50 ng/L MT处理组（P<0.01）和100 ng/L MT处理组（P<0.05）精巢dnmt3基因相对表达量显著（或极显著）降低;25 ng/L MT处理组和50 ng/L MT处理组精巢dnmt5基因相对表达量显著（P<0.05）降低;50 ng/L MT处理组精巢dnmt7基因相对表达量显著（P<0.05）降低。MT处理稀有鮈鲫雄鱼14 d,100 ng/L的MT处理组dnmt3、dnmt5、dnmt6、dnmt7基因相对表达量显著（P<0.05）升高;50 ng/L的MT处理组dnmt1基因相对表达量极显著（P<0.01）升高。MT处理稀有鮈鲫雄鱼21 d,50 ng/L MT处理组和100 ng/L MT处理组dnmt1基因相对表达量显著（P<0.05）升高;100 ng/L MT处理组dnmt8基因相对表达量极显著（P<0.01）升高。[结论] MT可以干扰稀有鮈鲫精巢DNA甲基转移酶基因dmmts的mRNA表达。MT对DNMTs酶活性的影响以及是否可以通过改变DNA甲基化水平对稀有鮈鲫精子的发生产生影响有待研究。

关键词: 17α-甲基睾酮, 稀有鮈鲫, 精巢, DNA甲基化转移酶基因

Abstract: [Objective] To investigate the effects of 17α-methyltestosterone （MT） on relative expression of DNA methyltransferase （DNMT） genes in testis of Gobiocypris rarus. [Method] Male Gobiocypris rarus were treated with varied doses of MT （25, 50, and 100 ng / L）. On the 7^th, 14^th and 21^st day post MT exposure, ten fishes were randomly selected from each MT treatment group, respectively, and they were dissected to collect testis tissues. The total RNA in the obtained testis tissues was extracted by one-step Trizol method, and the relative expression of DNA methyltransferase genes （dnmts） was detected by real-time quantitative PCR （qRT-PCR）. [Result] On 7^th day post MT exposure, compared with control group, the relative expression of dnmt3 gene in testis tissues in 25 ng/L （P<0.05）, 50 ng/L （P<0.01）, and 100 ng/L （P<0.05） MT treatment groups was significantly or extremely significantly reduced; in the testis tissues of 25 ng/L and 50 ng/L MT treatment groups, the relative expression of dnmt5 gene fell significantly （P<0.05）; 50 ng/L MT treatment group had significantly （P<0.05） decreased relative expression of dnmt7 gene in the testis tissues. On 14^th day post MT exposure, compared with control group, the relative expression of dnmt3, dnmt5, dnmt6, and dnmt7 genes increased significantly （P<0.05） in 100 ng/L MT treatment group, and that of dnmt1 gene increased extremely significantly （P<0.01） in 50 ng/L MT treatment group. On 21^st day post MT exposure, compared with control group, 50 ng/L and 100 ng/L MT treatment groups had significantly （P<0.05） increased relative expression of dnmt1 gene, and 100 ng/L MT treatment group had extremely significantly （P<0.01） higher relative expression of dnmt8 gene. [Conclusion] MT can interfere with the mRNA expression of dmmts genes in testis of Gobiocypris rarus. Further studies are expected to focus on the effects of MT on enzymatic activity of DNA methyltransferases, as well as on whether MT affects spermatogenesis of Gobiocypris rarus by altering DNA methylation levels.

Key words: 17α-methyltestosterone, Gobiocypris rarus, testis, DNA methyltransferase gene

中图分类号:

周俊亮, 王贤珍, 侯晓蕾, 杨琼, 陈越, 荣伟雅, 刘青, 王伟伟, 宋晶, 刘少贞. 17α-甲基睾酮对稀有鮈鲫精巢DNA甲基转移酶基因表达的影响[J]. 畜牧与饲料科学, 2022, 43(2): 1-6.

ZHOU Jun-liang, WANG Xian-zhen, HOU Xiao-lei, YANG Qiong, CHEN Yue, RONG Wei-ya, LIU Qing, WANG Wei-wei, SONG Jing, LIU Shao-zhen. Effects of 17α-methyltestosterone on Expression of DNA Methyltransferase Genes in Testis of Gobiocypris rarus[J]. Animal Husbandry and Feed Science, 2022, 43(2): 1-6.

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