摘要
【目的】克隆弓背青鳉(Oryzias curvinotus)雌激素受体基因(Estrogen receptor, er),分析在组织、胚胎发育时期中表达特征和雌二醇暴露后表达响应,为探究er介导雌激素调控机制提供理论依据。【方法】通过弓背青鳉肝脏转录组数据筛选进行er序列扩增,实时定量PCR(RT-q PCR)检测不同组织、不同胚胎发育时期和雌二醇暴露后的er表达量。【结果】3种ocuerα/erβ1/erβ2的编码区序列分别为1 860 bp、1 689 bp和2 007 bp,编码氨基酸个数相应为619、562和688。定量结果显示,3种er主要在肝脏和性腺高表达。胚胎发育过程中,ocuerα在卵裂期至囊胚期表达量低,在囊胚期后表达量逐渐上升持续到孵化;2种ocuerβ亚型均在胚胎发育初期有较高表达,随后ocuerβ1在原肠胚后开始降低,ocuerβ2在囊胚期后开始降低,在器官发育后期表达量均逐渐上升直至孵化。弓背青鳉仔鱼雌二醇暴露结果显示,各浓度雌二醇暴露均能使ocuerα和弓背青鳉卵黄蛋白基因(Vitellogenin,vtg)表达量上升,ocuerβ1仅在20μg/L上调,ocuerβ2表达量下调。【结论】ocuer不同亚型在介导雌激素调控中起着重要作用。
【Objective】To provide a theoretical basis for the mechanisms of estrogen receptor functions of Oryzias curvinotus, ocuer genes were cloned and analyzed. The tissue distribution and expression of embryonic development stage and response to estradiol were also evaluated. 【Method】The ocuer cDNA was amplified by the combination of transcriptome and reverse transcription PCR(RT-PCR).Tissue distribution of ocuer genes and expression of embryonic development stage and response to estradiol were analyzed by RT-qPCR. 【Result】The lengths of ocuerα/erβ1/erβ2 are 1971 bp, 2006 bp and 2365 bp, respectively, encoding of 619, 562 and 688 amino acids, correspondingly. The result of RT-qPCR showed that three ers were mainly expressed in the liver and gonads. The ocuerα was highly expressed in testis, ovary and female liver;ocuerβ1 was highly expressed in ovary and male liver;ocuerβ2 was highly expressed in gonad and liver. During the development of embryonic, the level of ocuerα expression was low, then increased gradually until hatching after blastocyst stage;both ocuerβsubtypes were highly expressed in the early stage of embryonic development, then decreased, but increased in the last stage of organ development until hatching. The exposure of E2 showed that the expression of ocuerα and ocuvtg increased at each concentration of E2 in the larvae of Oryzias curvinotu,and ocuerβ1 was only up-regulated at 20 μg/L but the ocuerβ2 was down-regulated. 【Conclusion】The ocuer gene play an important role in estrogen regulation.
作者
黄顺楷
郭昱嵩
汪淳
张海瑞
王淑如
董忠典
Huang Shun-kai;GUO Yu-song;WANG Chun;ZHANG Hai-rui;WANG Shu-ru;DONG Zhong-dian(Fisheries College,Guangdong Ocean University,Zhanjiang 524088,China)
出处
《广东海洋大学学报》
CAS
2019年第2期8-19,共12页
Journal of Guangdong Ocean University
基金
国家自然科学基金项目(41806195
31201996)
广东省人才项目(Yq2013089)
关键词
弓背青鳉
雌激素受体
雌激素
胚胎发育
Oryzias curvinotus
estrogen receptor
estrogen
embryonic development
