|
|
|
| Molecular Cloning, Sequence Structure and Expression Analysisof Gonadal Aromatase Gene in Puffer Takifugu obscures |
| GAO Yingying1,2, LIU Xinfu1,3, HU Peng4, HUANG Bin1,3, LIU Bin1,3 |
1.Yellow Sea Fisheries Research Institute, Chinese Academy of Fishery Sciences, Qingdao 266071, China;
2.College of Fisheries and Life Sciences, Shanghai Ocean University, Shanghai 201306, China;
3.Qingdao National Laboratory for Marine Science and Technology, Qingdao 266071, China;
4.Neijiang Normal University, Neijiang 641100, China |
|
|
|
|
Abstract The gonadal aromatase gene (cyp19a) was a kind of gene that regulates the sex hormone balance. The purpose of this study was to preliminarily investigate the role of gonadal aromatase gene in gonad development and sex differentiation in puffer Takifugu obscurus, and we successfully cloned the full-length cDNA sequence of cyp19a in puffer through RACE and Real-Time PCR technology. The full length of cyp19a sequence was 1786 bp, encoding 514 amino acids, and sharing the maximal homology with the genes of puffer T. rubripes and T. niphobles. The amino acid sequence analysis showed that the protein encoded by cyp19a was hydrophilic and stable protein, but there is no signal peptide sequence. There were 38 phosphorylation sites, 4 N-glycosylation sites, 2 transmembrane domains in the sequence. Meanwhile, the multiple amino acid sequence alignment showed that the amino acid sequence of gonadal aromatase was very conservative in both mammalian and fish, and contained some functional conservative regions such as the transmembrane region, I-helix region, Ozol′s peptide region, aromatase-specific conserved region and heme binding region. Furthermore, expression analysis showed that the cyp19a was highly abundant in ovary and muscle, but less abundant in other tissues. The expression of cyp19a in the ovary of juvenile fish at different developmental stages showed a trend of increasing first and then decreasing rapidly, without in the testis. The findings indicated that cyp19a may be involved in the development of ovarian after gonad differentiation, maintenance of female characteristics and development of other tissues in female puffer.
|
|
Received: 25 December 2018
|
|
|
|
|
|
[1]阳清发,怀向军,苏明,等.暗纹东方鲀的开发利用与管理[J].渔业现代化,2015,42(1):28-31.
[2]张迪,杨正勇,张英丽.中国河鲀养殖产业发展现状与发展对策[J].科学养鱼,2017(12):1-3.
[3]刘延正,徐承旭.河鲀“解禁”规模化养殖将成主流[J].水产科技情报,2017(5):284-285.
[4]华元渝,李廷友,邹宏海.养殖型暗纹东方鲀生长与发育特性[J].水产学报,2004,28(1):8-14.
[5]Coumailleau P, Pellegrini E, Adrio F, et al. Aromatase, estrogen receptors and brain development in fish and amphibians[J]. Biochimica Et Biophysica Acta—Gene Regulatory Mechanisms,2015,1849(2):152-162.
[6]Guiguen Y, Fostier A, Piferrer F, et al. Ovarian aromatase and estrogens:a pivotal role for gonadal sex differentiation and sex change in fish[J]. General and Comparative Endocrinology,2010,165(3):352-366.
[7]Piferrer F, Blazquez M. Aromatase distribution and regulation in fish[J]. Fish Physiology and Biochemistry,2005,31(2/3):215-226.
[8]张东茜,温海深.绿鳍马面鲀(Navodonsep tentrionalis) CYP19a基因克隆与表达特征分析[J].海洋与湖沼,2013,44(4):1086-1094.
[9]包海声.Cyp19a1和Foxl2基因在中华鳖早期卵巢分化中的功能研究[D].上海:上海海洋大学,2017.
[10]黄波,杨小玉,戴奇,等.芳香化酶抑制剂对暗纹东方鲀CYP19A、DMRT1基因表达及性腺分化的影响[J].中国水产科学,2013,20(1):68-74.
[11]李广丽,刘晓春,林浩然.芳香化酶抑制剂letrozole对赤点石斑鱼(Epinephelus akaara)性逆转的作用[J].生理学报,2005,57(4):473-479.
[12]邹立军,龚婧,吉璐,等.翘嘴鳜性腺型芳香化酶基因CYP19a的克隆及表达研究[J].生命科学研究,2017,21(4):295-301.
[13]甘为,方展强.食蚊鱼CYP19a基因的克隆与序列分析[J].水产学报,2016,40(10):1542-1555.
[14]陈芸,周鹏,张子平,等.大黄鱼cyp19a/b基因的克隆与表达分析[J].集美大学学报:自然科学版,2015,20(2):81-89.
[15]金国雄,温海深,柳学周,等.条斑星鲽CYP19a基因克隆及其在雄鱼生殖周期中的表达[J].水产学报,2010,34(2):194-203.
[16]邓思平.半滑舌鳎性别相关基因P450芳香化酶、FTZ-F1和DMRT1基因克隆及表达分析[D].青岛:中国海洋大学,2007.
[17]徐跑.黄颡鱼性别相关基因Sox9、Ftz-F1和P450arom的研究[D].南京:南京农业大学,2006.
[18]Deng S P, Chen S L, Xu J Y, et al. Molecular cloning, characterization and expression analysis of gonadal P450 aromatase in the half-smooth tongue-sole, Cynoglossusse milaevis[J]. Aquaculture,2009,287(1/2):211-218.
[19]Sawyer S J, Gerstner K A, Callard G V, et al. Real-time PCR analysis of cytochrome P450 aromatase expression in zebrafish: gene specific tissue distribution, sex differences, developmental programming and estrogen regulation[J]. General and Comparative Endocrinology,2006,147(2):108-117.
[20]陶敏,刘少军,张卓慧,等.不同倍性鲫鲤性腺型芳香化酶CYP19a1a基因cDNA的克隆及表达[J].水产学报,2014,38(9):1201-1210.
[21]Trant J M, Gavassa S, Ackers J, et al. Developmental expression of cytochrome P450 aromatase genes in zebrafish fry[J]. Journal of Experimental Zoology,2001,290(5):475-483.
[22]Barney M, Patil J G, Gunasekera R M, et al. Distinct cytochrome P450 aromatase isoforms in the common carp (Cyprinus carpio): sexual dimorphism and onset of ontogenic expression[J]. General and Comparative Endocrinology,2008,156(3):499-508.
[23]Chiang E F, Yan Y L, Tong S K, et al. Characterization of duplicated zebrafish CYP19 genes [J]. Journal of Experimental Zoology,2001,290(7):709-714.
[24]Chourasia T K, Joy K P. Ovarian P450 aromatase activity in the catfish Heteropneustes fossilis: seasonal changes and effects of catecholestrogens[J]. General and Comparative Endocrinology,2008,156(3):537-543. |
|
|
|
