| 施氏鲟促卵泡激素受体基因cDNA的克隆、表达及调控 |
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| 引用本文： | 高雪,韩世成,马波,吕伟华,王念民,张颖.施氏鲟促卵泡激素受体基因cDNA的克隆、表达及调控[J].淡水渔业,2020(3):56-64. |
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| 作者姓名： | 高雪 韩世成 马波 吕伟华 王念民 张颖 |
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| 作者单位： | 中国水产科学研究院黑龙江水产研究所;上海海洋大学农业农村部淡水种质资源重点实验室 |
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| 基金项目： | 中央级公益性科研院所基本科研业务费专项资金项目(HSY201609)。 |
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| 摘 要： | 为研究促卵泡激素受体基因(FSHR)在施氏鲟(Acipenser schrenckii)性腺分化过程中的表达、分布及调控作用,采用反转录聚合酶链式反应(RT-PCR)和RACE技术克隆得到施氏鲟促性腺激素(FSHR)基因的全长cDNA序列,并对其编码氨基酸序列的特征、基因表达及调控作用进行了分析。结果显示:施氏鲟FSHR基因的cDNA全长为2696 bp,编码661个氨基酸,属于含有信号肽、跨膜结构且分泌到细胞外的疏水性蛋白。氨基酸同源性及进化分析表明,施氏鲟与小体鲟(A.ruthenus)FSHR序列一致性最高,亲缘关系最近。PCR结果显示,FSHR mRNA的表达具有广泛性,各组织中均有表达,但在性腺、垂体中的表达量较高,显著高于其他组织中的表达量。LHRH-A注射实验显示:不同浓度组中,施氏鲟性腺组织中FSHR mRNA的表达量呈先升高后降低的变化趋势;与对照组相比,1μg/kg组在诱导3 d时性腺组织中FSHR mRNA的表达量达到最高,极显著高于3μg/kg和5μg/kg组。与性腺组织中FSHR mRNA表达模式不同,垂体中FSHR mRNA的表达呈升高后降低趋势,3μg/kg组在注射后第3天达最大值。综上结果表明,LHRH-A可通过调控FSHR的表达参与施氏鲟的早期性腺发育。
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| 关 键 词： | 施氏鲟(Acipenser schrenckii) 促性腺激素(FSHR) 基因克隆 序列分析 基因表达 |
Cloning,expression and regulation of follicle stimulating hormone receptor gene cDNA in Acipenser schrenckii |
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| GAO Xue,HAN Shi-cheng,MA Bo,LV Wei-hua,WANG Nian-min,ZHANG Ying.Cloning,expression and regulation of follicle stimulating hormone receptor gene cDNA in Acipenser schrenckii[J].Freshwater Fisheries,2020(3):56-64. |
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| Authors: | GAO Xue HAN Shi-cheng MA Bo LV Wei-hua WANG Nian-min ZHANG Ying |
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| Institution: | (Heilongjiang River Fisheries Research Institute,Chinese Academy of Fishery Sciences,Harbin 150070,China;Key Laboratory of Freshwater Fishery Germplasm Resource,Ministry of Agriculture and Rural Affairs,Shanghai Ocean University,Shanghai 201306,China) |
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| Abstract: | The purpose of this study was to investigate the expression,distribution and regulation of the FSHR gene in the gonadal differentiation of Acipenser schrenckii.The full-length cDNA sequence of the gene was cloned by RT-PCR and RACE.Meanwhile,the characteristics,spatiotemporal expression and regulation of its amino acid sequence were analysed.The results showed that the full length of the gene was 2696 bp,encoding 661 amino acids.It was a hydrophobic protein with a signal peptide,transmembrane structure and extracellular secretion.The analysis of amino acid homology and evolution showed that the FSHR sequence of A.ruthenus and A.sturgeon were the most consistent,and the genetic relationship was the closest.The results of PCR showed that the expression of FSHR mRNA was widespread in all tissues,but it was higher in gonad and pituitary than that in other tissues.LHRH-A injection experiment showed that the expression of FSHR mRNA in the gonad of A.schrenckii increased first and then decreased in different concentration groups.Compared with the control group,the expression of FSHR mRNA in the gonad of 1μg/kg group reached the highest level on the third day of induction,significantly higher than that of 3μg/kg and 5μg/kg groups.Different from the expression pattern of FSHR mRNA in the gonad,the expression of FSHR mRNA in pituitary increased and then decreased.The maximum value of FSHR mRNA in 3μg/kg group reached on the 3rd day after injection.In conclusion,LHRH-A can participate in the early gonadal development of A.schrenckii by regulating the expression of FSHR. |
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| Keywords: | Acipenser schrenckii FSHR gene cloning sequence analysis gene expression |
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