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何勇凤,朱永久,郭威,杨德国,吴兴兵.2016.四川裂腹鱼脑型芳香化酶Cyp19b基因组织分布及温度对其表达水平的影响.动物学杂志,51(2):281-290.
四川裂腹鱼脑型芳香化酶Cyp19b基因组织分布及温度对其表达水平的影响
Tissue Distribution of Brain Aromatase Gene Cyp19b in Schizothorax kozlovi and Temperature Effects on Its Expression
投稿时间:2015-07-22  修订日期:2016-02-15
DOI:DOI: 10.13859/j.cjz.201602014
中文关键词:  四川裂腹鱼  Cyp19b  组织表达  温度
英文关键词:Schizothorax kozlovi  Cyp19b  Tissue expression  Temperature
基金项目:中国水产科学研究院长江水产研究所所长基金,现代农业人才支撑计划(2130106)
作者单位E-mail
何勇凤 中国水产科学研究院长江水产研究所 heyf@yfi.ac.cn 
朱永久 中国水产科学研究院长江水产研究所  
郭威 中国水产科学研究院长江水产研究所  
杨德国 中国水产科学研究院长江水产研究所 yangdg@yfi.ac.cn 
吴兴兵 中国水产科学研究院长江水产研究所  
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中文摘要:
      为研究脑型芳香化酶基因Cyp19b在四川裂腹鱼(Schizothorax kozlovi)早期性别分化中的作用,采用RACE方法从四川裂腹鱼脑中扩增得到该基因cDNA全长序列,并应用荧光定量RT-PCR技术测定该基因mRNA的相对表达量,探讨该基因在不同规格四川裂腹鱼鳃、脑、心、肝、脾、肾、肌肉、精巢、卵巢组织中的表达差异,以及温度对其早期仔鱼阶段该基因表达的影响。四川裂腹鱼Cyp19b基因的cDNA全长序列共3 021 bp,共编码507个氨基酸,属于脑型芳香化酶基因;四川裂腹鱼Cyp19b基因编码的氨基酸序列与其他鱼类脑型芳香化酶基因编码的氨基酸序列同源性可达70%以上,而与性腺型芳香化酶基因编码的氨基酸序列同源性较低,为64%左右;四川裂腹鱼Cyp19b基因仅在脑组织中表达,具有比较高的组织特异性,且随着个体的增长,逐渐呈现显著的雌雄差异;对不同温度处理12日龄仔鱼6 d后,低温(10℃和14℃)能显著促进Cyp19b基因表达量的升高,但高温(26℃)却对其表达量无显著影响作用。由此推测,脑型芳香化酶基因Cyp19b可能在低温导致四川裂腹鱼雌性化过程中发挥着重要作用。
英文摘要:
      Previous studies by authors indicated that low temperature could result in more females of Schizothorax kozlovi during the early developmental stage. In order to elucidate the role of Cyp19b in sex differentiation of S. kozlovi, the full length cDNA of Cyp19b was cloned from the brain of S. kozlovi by using rapid amplification of cDNA ends (RACE) method. Relative mRNA expression levels of Cyp19b were determined by using quantitative real-time PCR (RT-PCR). Then the differences of Cyp19b mRNA expression levels in gills, brain, heart, liver, spleen, kidney, muscle, testis and ovary among different body-sized groups were discussed, while temperature effects on its expression during the larval stage of S. kozlovi were evaluated. The results indicated that: the full length cDNA of Cyp19b was 3 021 bp, which encoded 507 amino acids and belonged to brain aromatase gene; the deduced amino acid sequence of Cyp19b of S. kozlovi had more than 70% identity compared with brain aromatase genes of other teleost fishes, but only around 64% when compared with gonadal aromatase genes of other teleost fishes; the mRNA expression of Cyp19b of S. kozlovi only appeared in brain but not in other tissues, presenting relatively high tissue specificity; One-Way Analysis of Variance (ANOVA) revealed that the mRNA expression levels of Cyp19b were not significantly different between females and males in small body-sized samples (mean standard length was 12 cm), but gradually showed significant difference between females and males in large body-sized samples (mean standard length was 26 cm) (Fig. 3); larvae at 12 days after hatching were cultured in five different temperature groups, ANOVA revealed that six days later the mRNA expression levels of Cyp19b were significantly increased by low temperature (10℃ and 14℃), but not influenced by high temperature (26℃) (Fig. 4). The present results suggested that brain aromatase gene Cyp19b may play important roles in the feminization of S. kozlovi affected by low temperature during the early developmental stage.
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