雌性三角帆蚌Srd5a1基因的分子鉴定和功能研究  被引量:1

Molecular identification and functional study of Srd5a1 in female Hyriopsis cumingii

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作  者:曹慕莲 霍滢朵 刘宗雨 缪煜琳 金鑫[1] 汪桂玲[1,2,3] CAO Mulian;HUO Yingduo;LIU Zongyu;MIAO Yulin;JIN Xin;WANG Guiling(Key Laboratory of Freshwater Aquatic Genetic Resources,Ministry of Agriculture and Rural Affairs,Shanghai Ocean University,Shanghai 201306,China;Shanghai Engineering Research Center of Aquaculture,Shanghai Ocean University,Shanghai 201306,China;Shanghai Collaborative Innovation for Aquatic Animal Genetics and Breeding,Shanghai Ocean University,Shanghai 201306,China)

机构地区:[1]上海海洋大学农业农村部淡水水产种质资源重点实验室,上海201306 [2]上海海洋大学上海水产养殖工程技术研究中心,上海201306 [3]上海海洋大学水产动物遗传育种中心上海市协同创新中心,上海201306

出  处:《上海海洋大学学报》2024年第2期297-307,共11页Journal of Shanghai Ocean University

基  金:国家重点研发计划“蓝色粮仓科技创新”项目(2022YFD2400700);国家自然科学基金(31772835);财政部和农业农村部国家现代农业产业技术体系项目(CAR-49)。

摘  要:性类固醇激素与生物生殖、配子排放以及性腺发育密切相关,Srd5a1基因在性类固醇激素合成过程中扮演重要角色。为了探究性类固醇激素合成相关酶基因Srd5a1对雌性三角帆蚌卵巢发育的影响,应用RACE克隆得到了该基因全长cDNA序列,通过实时荧光定量(qRT-PCR)、原位杂交技术以及不同浓度17β-雌二醇(E2)和17α-甲基睾酮(MT)激素处理来探究基因的表达特性。结果显示,Srd5a1 cDNA全长2224 bp,包括446 bp 5′-UTR、953 bp 3′-UTR和825 bp ORF,编码274个氨基酸。Srd5a1基因在性腺中的表达呈二态性,在卵巢中表达量更高;且在卵巢排放期的表达量最高,极显著高于其他各时期。原位杂交结果显示,Srd5a1基因在卵巢卵母细胞、卵泡以及精巢精母细胞中有表达。不同浓度E2和MT处理24 d后,Srd5a1基因的表达发生了变化。综上,Srd5a1基因可能在雌性三角帆蚌卵巢卵母细胞发育成熟和配子排放中发挥一定作用,对探索三角帆蚌性腺生长发育具有重要意义,同时为实现三角帆蚌单性化养殖提供理论参考价值。Sex steroid hormones are closely related to biological reproduction,gamete emission and gonadal development.And Srd5a1 plays an important role in the synthesis of sex steroid hormones.In order to investigate the effect of the sex steroid hormone synthesis-related enzyme gene Srd5a1 on ovarian development in female Hyriopsis cumingii,Srd5a1 was obtained from the gonads of H.cumingii by RACE(Rapid-amplificiantion of cDNA ends).The expression characteristics of Srd5a1 were investigated by realtime fluorescence quantification(qRT-PCR),in situ hybridization and in vitro induction of 17β-estradiol(E2)and 17α-methyltestosterone(MT)at different concentrations for 24 days.The results showed that the full-length cDNA of Srd5a1 was 2224 bp,including ORF 825 bp,3′-UTR 953 bp,and 5′-UTR 446 bp,encoding 274 amino acids.The expression of Srd5a1 in the gonad was bimodal,with higher expression in the ovary than that in the testis;Srd5a1 had the highest expression level in the ovarian discharge period,which was extremely significantly higher than that in all other periods.In situ hybridization results showed that Srd5a1 detected positive signals in ovarian oocytes and follicles as well as spermatocytes of the testis.The expression of Srd5a1 was changed after 24 days of treatment with different concentrations of E2 and MT.In conclusion,Srd5a1 may play a role in the developmental maturation of the ovary oocyte and gamete emission in the female H.cumingii.This study is important for exploring the gonadal growth and development of H.cumingii,and also provides a theoretical reference value for the realization of monogamous breeding of H.cumingii.

关 键 词:三角帆蚌 Srd5a1基因 卵巢发育 17Β-雌二醇 17α-甲基睾酮 

分 类 号:S917.4[农业科学—水产科学]

 

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