2种乌贼雌激素相关受体基因的克隆与表达特性分析
|
摘要
雌激素是重要的类固醇类激素,在软体动物中,雌激素一般不能与雌激素受体识别并结合,但却能通过调控一类孤儿核受体——雌激素相关受体(estrogen-related receptor,ERR)的表达而发挥其生殖调控的部分功能,但有关ERR在软体动物中的确切生物学特性及功能还知之甚少。本研究首次从2种海洋头足类:曼氏无针乌贼(Sepiella japonica)和白斑乌贼(Sepia latimanus)中,克隆得到了ERR基因的全序列,并对其在乌贼不同组织及不同发育时期的表达特征进行研究。结果表明:乌贼中仅克隆得到了1种ERR基因亚型(ERRγ),在曼氏无针乌贼和白斑乌贼中的全长分别为1 513 bp和1 547 bp,两者分别编码339和351个氨基酸残基,这些氨基酸可形成A/B、C、D和E 4个典型的核受体结构域。进化树分析表明,乌贼的ERR基因C结构域和E结构域较其他软体动物相对高度保守,曼氏无针乌贼和白斑乌贼ERR基因与同为头足类的双斑蛸(Octopus bimaculoides)的ERR同源性最高,分别达到95%和85%。2种乌贼ERR基因在不同组织中表达谱相仿,在不同组织中广泛表达,但在脑、肝、卵巢等生殖相关器官中表达量最高。2种乌贼卵巢中的ERR基因表达量都随性成熟时间的推进而逐渐上调,而在繁殖期后显著下调(P<0.05)。该结果表明乌贼的ERR基因可能在其生殖发育和性成熟过程中起着重要的调控作用。[中国渔业质量与标准,2019,9(2):16-23]
Estrogen is an important steroid hormone in animals. In mollusk, estrogen can′t bind to its natural estrogen receptor(ER) to activate the estrogen function, nevertheless, it can regulate expression of a type of orphan nuclear receptor, called estrogen related receptor(ERR), to partly activate estrogen pathway in reproduction regulation. However, the function and the signaling pathways of ERR in mollusk have not yet been clearly elucidated. In this study, a full-length gene sequence encoding the ERR was firstly cloned from two cuttlefish, Sepiella japonica and Sepia latimanus. The expression profiles of the ERR in various tissues were also studied during different development period. The subtype of ERR(EERγ) cDNA clone was found to contain 1 513 bp and 1 547 bp nucleotides including an open-reading frame encoding 339(SjEER) and 351(SlEER) amino acid(aa) residues in Sepiella japonica and Sepia latimanus, respectively. The deduced ER protein consisted of four nuclear receptor characteristic domains of A/B,C,D and E. Based on a phylogenetic analysis, the EER DNA-binding domain and ligand-binding domain were highly conserved compared to other mollusces. SjEER and SlEER were found to have the highest aa identities to common octopus(Octopus bimaculoides) EER with an identity of 95% and 85%, respectively. Tissue expression analysis confirmed that EER was widely distributed among various tissues and predominantly expressed in the brain, liver, gonad(ovary). The EER expression was markedly up regulated in the ovary during the early sexual maturation period and down regulated after spawning in two cuttlefishes(P<0.05). These results suggest that EER may play an important role in regulating the reproductive cycle of cuttlefish of S.japonica and S.latimanus.[Chinese Fishery Quality and Standards,2019,9(2):16-23]
Estrogen is an important steroid hormone in animals. In mollusk, estrogen can′t bind to its natural estrogen receptor(ER) to activate the estrogen function, nevertheless, it can regulate expression of a type of orphan nuclear receptor, called estrogen related receptor(ERR), to partly activate estrogen pathway in reproduction regulation. However, the function and the signaling pathways of ERR in mollusk have not yet been clearly elucidated. In this study, a full-length gene sequence encoding the ERR was firstly cloned from two cuttlefish, Sepiella japonica and Sepia latimanus. The expression profiles of the ERR in various tissues were also studied during different development period. The subtype of ERR(EERγ) cDNA clone was found to contain 1 513 bp and 1 547 bp nucleotides including an open-reading frame encoding 339(SjEER) and 351(SlEER) amino acid(aa) residues in Sepiella japonica and Sepia latimanus, respectively. The deduced ER protein consisted of four nuclear receptor characteristic domains of A/B,C,D and E. Based on a phylogenetic analysis, the EER DNA-binding domain and ligand-binding domain were highly conserved compared to other mollusces. SjEER and SlEER were found to have the highest aa identities to common octopus(Octopus bimaculoides) EER with an identity of 95% and 85%, respectively. Tissue expression analysis confirmed that EER was widely distributed among various tissues and predominantly expressed in the brain, liver, gonad(ovary). The EER expression was markedly up regulated in the ovary during the early sexual maturation period and down regulated after spawning in two cuttlefishes(P<0.05). These results suggest that EER may play an important role in regulating the reproductive cycle of cuttlefish of S.japonica and S.latimanus.[Chinese Fishery Quality and Standards,2019,9(2):16-23]
引文
[1] Lv Z M,Liu W,Liu L Q,et al.Cloning,characterization,and expression profile of estrogen receptor in common Chinese cuttlefish,Sepiella japonica[J].J Exp Zool,2016,325A:181-193.
[2] Di Cosmo A,Cristo C D,Paolucci M.Sex steroid hormone fluctuationsand morphological changes of the reproductive system ofthe female of Octopus vulgaris throughout the annual cycle[J].J Exp Zool,2001,289:33-47.
[3] Gauthier-Clerc S,Pellerin J,Amiard J C.Estradiol-17β andtestosterone concentrations in male and female Mya arenaria (Mollusca bivalvia) during the reproductive cycle[J].Gen Comp Endocrinol,2006,145:133-139.
[4] Croll R P,Wang C.Possible roles of sex steroids in the control of reproduction in bivalve mollusks[J].Aquaculture,2007,272:76-86.
[5] Osada M,Takamura T,Sato H,et al.Vitellogenin synthesisin the ovary of scallop,Patinopecten yessoensis:control by estradiol-17 beta and the central nervous system[J].J Exp Zool,2003,299A:172-179.
[6] Baker M E,Chandsawangbhuwana C.Analysis of 3D modelsof octopus estrogen receptor with estradiol:evidence for stericclashes that preventestrogen binding[J].BiochemBiophys Res Commun,2007,361:782-788.
[7] Giguere V,Yang N,Segui P,et al.Identification of a new class of steroid hormone receptors[J].Nature,1999,331(6151):91-94.
[8] Tennessen J M,Baker K D,Lam G,et al.The drosophila estrogen-related receptordirects a metabolic switchthat supports developmental growth[J].Cell Metab,2011,13:139-148.
[9] Gibson D A,Saunders P T K.Estrogen dependent signaling in reproductive tissues – A role for estrogen receptors and estrogen related receptors[J].Mol Cell Endocrinol,2012,348:361-372.
[10] Mitsunaga K,Araki K,Mizusaki H,et al.Loss of PGC- specific expression of the orphan nuclear receptor ERRβ results in reduction of germ cell number in mouse embryos[J].Mech Mater,2004,121(3):237-246.
[11] Park E,Kumar S,Lee B,et al.Estrogen receptor-related receptor g regulates testicular steroidogenesis through direct and indirect regulation of steroidogenic gene expression[J].Mol Cell Endocrinol,2017,452:15-24.
[12] 刘英华,何宁,姜淑卿,等.雌激素相关受体α与雌激素受体α通路相关性研究[J].中华肿瘤防治杂志,2013,20(17):1306-1314.
[13] 董正之.中国动物志:软体动物门,头足纲[M].北京:科学出版社,1988.
[14] 董正之.世界海洋经济头足类生物学[M].济南:山东科学技术出版社,1991.
[15] 陈健,周度金.雌激素受体相关受体研究进展[J].生命的化学,2001,21(6):487-489.
[16] Lv Z M,Liu W,Liu L Q,et al.De novo assembly and comparison of the ovarian transcriptomes of thecommon Chinese cuttlefish (Sepiella japonica) with different gonadal development[J].Genomics Data,2016,7:155–168.
[17] 欧阳霞辉,侯兰新,高丹丹,等.拟黑多刺蚁(Polyrhachis vicina)性腺雌激素相关受体基因的表达[J].西北民族大学学报,2014,35(1):53-57.
[18] He H,Xi G S,Lu X.Molecular cloning,characterization,and expression analysisof an estrogen receptor-related receptor homologue in the cricket,Teleogryllus emma[J].J Insect Sci,2010,10(188):1-16.
[19] Cheung N K M,Cheung A C K,Ye R R.Expression profile of oestrogen receptors and oestrogen-related receptors is organ specific and sex dependent:the Japanese medaka Oryzias latipes model[J].J Fish Biol,2013,83:295-310.
[20] Tarrant A M,Greytak S R,Callard G V,et al.Estrogen receptor-related receptors in the killifish Fundulus heteroclitus:Diversity,expression,and estrogen responsiveness[J].J Molec Endocrinol,2006,37(1):105-120.
[21] Bannister R,Beresford N,May D,et al.Novel estrogen receptor-related Transcripts in Marisa cornuarietis;a freshwater snail with reported sensitivity to estrogenic chemicals[J].Environ Sci Technol,2007,41(7):2643-2650.
[22] Nagasawa K,Treenc N,Kondo R,et al.Molecular characterization of an estrogen receptor and estrogen -related receptor and their autoregulatory capabilities in two Mytilus species[J].Gene,2015,564:153-159.
[23] Ouyang X H,Xi G S,Bu C P,et al.Molecular cloning and expression of an estrogen receptor-related receptor gene in the ant Polyrhachis vicina (Hymenoptera:Formicidae)[J].Ann Entomol Soc Am,2009,102(2):295-302.
[2] Di Cosmo A,Cristo C D,Paolucci M.Sex steroid hormone fluctuationsand morphological changes of the reproductive system ofthe female of Octopus vulgaris throughout the annual cycle[J].J Exp Zool,2001,289:33-47.
[3] Gauthier-Clerc S,Pellerin J,Amiard J C.Estradiol-17β andtestosterone concentrations in male and female Mya arenaria (Mollusca bivalvia) during the reproductive cycle[J].Gen Comp Endocrinol,2006,145:133-139.
[4] Croll R P,Wang C.Possible roles of sex steroids in the control of reproduction in bivalve mollusks[J].Aquaculture,2007,272:76-86.
[5] Osada M,Takamura T,Sato H,et al.Vitellogenin synthesisin the ovary of scallop,Patinopecten yessoensis:control by estradiol-17 beta and the central nervous system[J].J Exp Zool,2003,299A:172-179.
[6] Baker M E,Chandsawangbhuwana C.Analysis of 3D modelsof octopus estrogen receptor with estradiol:evidence for stericclashes that preventestrogen binding[J].BiochemBiophys Res Commun,2007,361:782-788.
[7] Giguere V,Yang N,Segui P,et al.Identification of a new class of steroid hormone receptors[J].Nature,1999,331(6151):91-94.
[8] Tennessen J M,Baker K D,Lam G,et al.The drosophila estrogen-related receptordirects a metabolic switchthat supports developmental growth[J].Cell Metab,2011,13:139-148.
[9] Gibson D A,Saunders P T K.Estrogen dependent signaling in reproductive tissues – A role for estrogen receptors and estrogen related receptors[J].Mol Cell Endocrinol,2012,348:361-372.
[10] Mitsunaga K,Araki K,Mizusaki H,et al.Loss of PGC- specific expression of the orphan nuclear receptor ERRβ results in reduction of germ cell number in mouse embryos[J].Mech Mater,2004,121(3):237-246.
[11] Park E,Kumar S,Lee B,et al.Estrogen receptor-related receptor g regulates testicular steroidogenesis through direct and indirect regulation of steroidogenic gene expression[J].Mol Cell Endocrinol,2017,452:15-24.
[12] 刘英华,何宁,姜淑卿,等.雌激素相关受体α与雌激素受体α通路相关性研究[J].中华肿瘤防治杂志,2013,20(17):1306-1314.
[13] 董正之.中国动物志:软体动物门,头足纲[M].北京:科学出版社,1988.
[14] 董正之.世界海洋经济头足类生物学[M].济南:山东科学技术出版社,1991.
[15] 陈健,周度金.雌激素受体相关受体研究进展[J].生命的化学,2001,21(6):487-489.
[16] Lv Z M,Liu W,Liu L Q,et al.De novo assembly and comparison of the ovarian transcriptomes of thecommon Chinese cuttlefish (Sepiella japonica) with different gonadal development[J].Genomics Data,2016,7:155–168.
[17] 欧阳霞辉,侯兰新,高丹丹,等.拟黑多刺蚁(Polyrhachis vicina)性腺雌激素相关受体基因的表达[J].西北民族大学学报,2014,35(1):53-57.
[18] He H,Xi G S,Lu X.Molecular cloning,characterization,and expression analysisof an estrogen receptor-related receptor homologue in the cricket,Teleogryllus emma[J].J Insect Sci,2010,10(188):1-16.
[19] Cheung N K M,Cheung A C K,Ye R R.Expression profile of oestrogen receptors and oestrogen-related receptors is organ specific and sex dependent:the Japanese medaka Oryzias latipes model[J].J Fish Biol,2013,83:295-310.
[20] Tarrant A M,Greytak S R,Callard G V,et al.Estrogen receptor-related receptors in the killifish Fundulus heteroclitus:Diversity,expression,and estrogen responsiveness[J].J Molec Endocrinol,2006,37(1):105-120.
[21] Bannister R,Beresford N,May D,et al.Novel estrogen receptor-related Transcripts in Marisa cornuarietis;a freshwater snail with reported sensitivity to estrogenic chemicals[J].Environ Sci Technol,2007,41(7):2643-2650.
[22] Nagasawa K,Treenc N,Kondo R,et al.Molecular characterization of an estrogen receptor and estrogen -related receptor and their autoregulatory capabilities in two Mytilus species[J].Gene,2015,564:153-159.
[23] Ouyang X H,Xi G S,Bu C P,et al.Molecular cloning and expression of an estrogen receptor-related receptor gene in the ant Polyrhachis vicina (Hymenoptera:Formicidae)[J].Ann Entomol Soc Am,2009,102(2):295-302.