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Cloning, Sequence Features and Expression Analysis of 17β -HSD3, A Key Gene Involved in Gonadal Development of Mulard Ducks (Cairina moschata) |
MIAO Zhong-Wei1,2, LI Li1,2*, ZHU Zhi-Ming1,2, ZHANG Lin-Li1,2, HAO Xiao-Na1,3, HUANG Qin-Lou1,2, ZHENG Nen-Zhu1,2, XIN Qing-Wu1,2* |
1 Institute of Animal Husbandry and Veterinary Medicine, Fujian Academy of Agricultural Sciences, Fuzhou 350013, China; 2 Fujian Key Laboratory of Animal Genetics and Breeding, Fuzhou 350013, China; 3 College of Animal Science (College of Beeology), Fujian Agriculture and Forestry University, Fuzhou 350002, China |
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Abstract The 17β-hydroxysteroid dehydrogenase (17β-HSD) eventually oxidizes androstenedione converted from cholesterol to testosterone, which is essential for testosterone synthesis and male reproductive traits. 17β- HSD3 was found to be involved in the pre-embryonic gonadal development of the Mulard duck (Cairina moschata) through the steroid hormone synthesis pathway. In this study, the full-length sequence of 17β-HSD3 gene was obtained by rapid amplification of cDNA ends (RACE) cloning method, sequence characteristics and physicochemical properties of proteins were predicted using bioinformatics analysis, and phylogenetic trees were constructed. The qPCR was utilized to analyze the tissue-specific expression of 17β -HSD3 gene in Mulard duck (brain, heart, liver, lung, muscle, gonad, kidney) and differential expression in the gonads of Mulard ducks, male parent Muscovy and female parent Pekin ducks. The results showed that the full-length sequence of 17β-HSD3 gene was 1 332 bp in Mulard duck, containing a 963 bp of open reading frame, and the length of 5' and 3' non-coding regions were 26 bp and 343 bp, respectively. The encoded protein contained 320 amino acids, which was an unstable protein. Homology and phylogenetic tree phylogenetic analysis showed that the 17β-HSD3 gene in Mulard duck have the closest relationship with swans (Cygnus olor). Tissue-specific expression analysis revealed that the 17β-HSD3 gene was highly significantly more expressed in the gonads of Mulard ducks than in other tissues (P<0.01), suggesting that 17β -HSD3 gene might be involved in gonadal development in Mulard duck. The expression of the 17β -HSD3 gene was significantly higher in Muscovy ducks and Pekin ducks with normal reproductive traits than in Mulard ducks (P<0.01), revealing that 17β - HSD3 gene might be associated with sterility in Mulard ducks. The study provides a theoretical basis for the function of the 17β -HSD3 gene in gonadal development, germ cell differentiation and other reproductive processes in the Mulard duck.
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Received: 27 June 2022
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Corresponding Authors:
* xinqingwu6656@126.com; lily102524@163.om
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[1] 曹体婷, 赵宗胜, 刘贤侠, 等. 2009. 鹌鹑及与鸡的杂交后代3β-HSD 和 P-450c17 基因的 cDNA 部分序列克隆[J]. 石河子大学学报(自然科学版), 27(04): 445-448. (Cao T T, Zhao Z S, Liu X X, et al. 2009. The Partial cDNA se-quences cloning of 3β -HSD and P-450c17 genes in Gal-lus, genetic hybrids of chicken-quail[J]. Journal of Shi-hezi University (Natural Science), 27(04): 445-448. ) [2] 李丽, 章琳俐, 缪中纬,等. 2021. 半番鸭卵巢发育不全关键基因的筛选与差异基因表达分析[J]. 农业生物技术学报, 29(9): 1761-1773. (Li L, Zhang L L, Miao Z W, et al. 2021. Screening of key genes for ovarian hypoplasia and analysis of differential gene expression in Mulard Ducks (Cairina moschata). 农业生物技术学报, 29(9): 1761-1773. ) [3] 李艳红. 2015. 睾丸酮丛毛单胞菌 LysR 基因对 3,17β-羟基类固醇脱氢酶的调控分析[D]. 硕士学位论文, 长春理工大学, 导师 : 于源华, pp, 22-36. (Li Y H. 2015. LysR gene on analysis of the regulation of 3, 17 hydroxyster-oid dehydrogenase in Comamonas testosterone[D]. Mas-Thesis for M. S., Changchun University of Science and Technology. Supervisor: Yu Y H, pp. 22-36 ) [4] 梁冬冬, 范兆飞, 邹玉霞, 等. 2017. 牙鲆 17β-HSD1 基因克隆及其表达调控的初步研究[J]. 海洋科学, 41(09): 65-73. (Liang D D, Fan Z F, ZOU Y X, et al. 2017. Molecu-lar characterization, expression, and regulation of 17β-HSD1 in the olive flounder Paralichthys olivaceus[J]. Marine Sciences, 41(09): 65-73. ) [5] 曾臻, 倪健斌, 谭强来, 等. 2020. 福建牡蛎 17β-HSD 基因的克隆及其生殖周期表达[J]. 应用海洋学学报, 39(1): 12-19. (Zeng Z, Ni J B, Tan Q L, et al. 2020. Cloning of 17β-HSD gene and its characterization in the Fujian oys-ter, Crassostrea angulata, during gonad development[J]. Journal of Applied Oceanography, 39(1): 12-19. ) [6] 张哲, 郑清莲, 王宏竹, 等. 2019. 17β-HSD3 在 SD 大鼠肾脏中的表达及其功能[J]. 华中科技大学学报(医学版), 48(06): 686-689. (Zhang Z, Zheng Q L, Wang H Z, et al. 2019. Expression and function of 17β -HSD3 in SD rat kidney[J]. Acta Medicinae Universitatis Scientiae et Technologiae Huazhong[J]. 48(06): 686-689. ) [7] 周林燕. 2004. 罗非鱼三种 17β-羟类固醇脱氢酶(17β-HSD1, 17β-HSD3, 17β-HSD8)的克隆、表达及酶活性鉴定[D]. 西南师范大学. 硕士学位论文, 导师: 王德寿, pp. 12-30. (Zhou L Y. 2004. Molecular cloning, gene expres-sion and enzyme activity charaeterization of three types of 17β-hydorxysteorid dehydorgenases (17β-HSDI, 17β-HSD3, 17β-HSD8) from Nile TilaPia, Oreochromis niloti-cus[D]. Southwest University. Master's thesis, Supervi-sor: Wang D S, pp. 12-30. ) [8] Agarwal D, Gireesh-Babu P, Pavan-Kumar A, et al. 2020. Mo-lecular characterization and expression profiling of 17-beta-hydroxysteroid dehydrogenase 2 and spermatogen-esis associated protein 2 genes in endangered catfish, Clarias magur (Hamilton, 1822)[J]. Animal Biotechnolo-gy, 31(2): 93-106. [9] Ben Rhouma B, Belguith N, Mnif M F, et al. 2013. A novel nonsense mutation in HSD17B3 gene in a Tunisian pa-tient with sexual ambiguity[J]. The Journal of Sexual Medicine, 10(10): 2586-2589. [10] Ben Rhouma B, Kallabi F, Mahfoudh N, et al. 2017. Novel cases of Tunisian patients with mutations in the gene en-coding 17β -hydroxysteroid dehydrogenase type 3 and a founder effect[J]. Journal of Steroid Biochemistry and Molecular Biology, 165(Pt A): 86-94. [11] Bhat I A, Rather M A, Nazir M I, et al. 2018. Cloning, charac-terisation, docking and expression analysis of 3-beta-hy-droxysteroid dehydrogenase during ontogenetic develop-ment and annual reproductive cycles in catfish (Clarias batrachus)[J]. Theriogenology, 105(1): 34-44. [12] Cheng Y, Yang Y, Wu Y, et al. 2020. The curcumin derivative, h10, suppresses hormone-dependent prostate cancer by inhibiting 17β -hydroxysteroid dehydrogenase type 3[J]. Frontiers in Pharmacology, 11: 637. [13] Ciftci N, Kayaş L, Çamtosun E, et al. 2022. 46, XY sex devel-opment defect due to a novel homozygous (splice site) c. 673_1g>c variation in the HSD17B3 gene: Case Report[J]. Journal of Clinical Research in Pediatric Endocrinol-ogy. 14(2): 233-238. [14] Cohen-Kettenis P T. 2005. Gender change in 46, XY persons with 5alpha-reductase-2 deficiency and 17beta-hy-droxysteroid dehydrogenase-3 deficiency[J]. Archives of Sexual Behaviour, 34(4): 399-410. [15] Cortés-Benítez F, Roy J, Perreault M, et al. 2019. A-and D-ring structural modifications of an androsterone deriva-tive inhibiting 17β -hydroxysteroid dehydrogenase type 3: Chemical synthesis and structure-activity relation-ships[J]. Journal of Medicinal Chemistry, 62(15): 7070-7088. [16] Day J M, Tutill H J, Purohit A, et al. 2008. Design and valida-tion of specific inhibitors of 17beta-hydroxysteroid de-hydrogenases for therapeutic application in breast and prostate cancer, and in endometriosis[J]. Endocrine Re-lated Cancer, 15(3): 665-692. [17] Engeli R T, Rhouma B B, Sager C P, et al. 2016. Biochemical analyses and molecular modeling explain the functional loss of 17β -hydroxysteroid dehydrogenase 3 mutant G133R in three Tunisian patients with 46, XY disorders of sex development[J]. Journal of Steroid Biochemistry and Molecular Biology. 155(Pt A): 147-154. [18] Engeli R T, Tsachaki M, Hassan H A, et al. 2017. Biochemical analysis of four missense mutations in the HSD17B3 gene associated with 46, xy disorders of sex develop-ment in Egyptian patients[J]. The Journal of Sexual Medicine, 14(9): 1165-1174. [19] Furtado P S, Moraes F, Lago R, et al. 2012. Gender dysphoria associated with disorders of sex development[J]. Nature Reviews Urology,9(11): 620-627. [20] Grimbly C, Caluseriu O, Metcalfe P, et al. 2016. 46,XY disor-der of sex development due to 17-beta hydroxysteroid dehydrogenase type 3 deficiency: A plea for timely ge-netic testing[J]. Journal of Pediatric Endocrinology & Metabolism. 2016: 12 [21] Khattab A, Yuen T, Yau M, et al. 2015. Pitfalls in hormonal di-agnosis of 17-beta hydroxysteroid dehydrogenase Ⅲ de-ficiency[J]. Journal of Pediatric Endocrinology & Me-tabolism, 28(5-6): 623-628. [22] Kirk C L, Lei Z M, Rao V, et al. 2004. De novo testosterone production in luteinizing hormone receptor knockout mice after transplantation of leydig stem cells[J]. Endo-crinology, 145(9): 4011-4015. [23] Kothandapani A, Jefcoate C R, Jorgensen J S. 2021. Choles-terol contributes to male sex differentiation through its developmental role in androgen synthesis and hedgehog signaling[J]. Endocrinology, 162(7): bqab066. [24] Li M, Xiong G, Maser E. 2013. A novel transcriptional repres-sor PhaR for the steroid-inducible expression of the 3,17β -hydroxysteroid dehydrogenase gene in Comamonas testosteroni ATCC11996[J]. Chemico-Biological Interac-tions, 202(1-3): 116-125. [25] Li X, Wang Z, Jiang Z, et al. 2016. Regulation of seminifer-ous tubule-associated stem leydig cells in adult rat testes[J]. Proceedings of The National Academy of Sciences of the USA, 113(10): 2666-2671. [26] Lukacik P, Kavanagh K L, Oppermann U. 2006. Structure and function of human 17beta-hydroxysteroid dehydrogenas-es[J]. Molecular and Cellular Endocrinology, 248(1-2): 61-71. [27] Marchais-Oberwinkler S, Henn C, Möller G, et al. 2011. 17β-hydroxysteroid dehydrogenases (17β -HSDs) as thera-peutic targets: Protein structures, functions, and recent progress in inhibitor development[J]. Journal of Steroid Biochemistry and Molecular Biology, 125(1-2): 66-82. [28]Mendonca B B, Gomes N L, Costa E M, et al. 2017. 46, XY disorder of sex development (DSD) due to 17β -hy-droxysteroid dehydrogenase type 3 deficiency[J]. Jour-nal of Steroid Biochemistry and Molecular Biology, 165(Pt A): 79-85. [29] Ning X, Yang Y, Deng H, et al. 2017. Development of 17β-hy-droxysteroid dehydrogenase type 3 as a target in hor-mone-dependent prostate cancer therapy[J]. Steroids, 121(5): 10-16. [30] Ohno S, Honda Y, Nakajima Y, et al. 2006. Sequence of pig 17beta-hydroxysteroid dehydrogenase type3 cDNA and its expression in mammalian cells[J]. Journal of Steroid Biochemistry and Molecular Biology, 01(2-3): 145-150. [31] Raghuveer K, Senthilkumaran B. 2012. Cloning and expres-sion of 3β-hydroxysteroid dehydrogenase during gonad-al recrudescence and after hCG induction in the air-breathing catfish (Clarias gariepinus)[J]. Steroids, 77(11):1133-1140. [32] Rajakumar A, Senthilkumaran B. 2014. Molecular cloning and expression analysis of 17b-hydroxysteroid dehydro-genase 1 and 12 during gonadal development, recrudes-cence and after in vivo hCG induction in catfish, Clarias batrachu[J]. Steroids, 92(12): 81-89. [33] Tang Y, Chen J Y, Ding G H, et al. 2021. Analyzing the gonad-al transcriptome of the frog Hoplobatrachus rugulosus to identify genes involved in sex development[J]. BMC Genomics, 22(1): 552. [34] Vicker N, Bailey H V, Day J M, et al. 2021. Substituted aryl benzylamines as potent and selective inhibitors of 17β -hydroxysteroid dehydrogenase type 3[J]. Molecules, 26(23): 7166. [35] Wang C L, Ying S J, Wang Z Y, et al. 2013. Molecular cloning and expression of 17β -hydroxysteroid dehydrogenase type 2 gene in Hu sheep[J]. Molecular Biology Reports, 40(2): 1073-1080. [36] Wu Y, Huang P, Xiong G, et al. 2015. Identification and isola-tion of a regulator protein for 3, 17β -HSD expressional regulation in Comamonas testosteroni[J]. Chemico-Bio-logical Interactions, 234(6): 197-204. [37] Xiao L, Bei Y, Li J, et al. 2021. Preclinical pharmacokinetics, tissue distribution and primary safety evaluation of a nov-el curcumin analogue h10 suspension, a potential 17β hy-droxysteroid dehydrogenase type 3 inhibitor[J]. Chemi-cal & Pharmaceutical Bulletin (Tokyo), 69(1): 52-58. [38] Xie W, Xia Q, Chen L, et al. 2021. Cloning and identification of a new repressor of 3, 17β -hydroxysteroid dehydroge-nase of comamonas testosteroni[J]. Molecular Biology Reports, 48(11): 7067-7075. [39] Xiong G, Maser E. 2015. Construction of a biosensor mutant of Comamonas testosteroni for testosterone determination by cloning the EGFP gene downstream to the regulatory region of the 3, 17β -HSD gene[J]. Chemico-Biological Interactions, 234(6): 188-196. [40] Yuan K, Zhao B, Li XW, et al. 2012. Effects of phthalates on 3β -hydroxysteroid dehydrogenase and 17β -hydroxyster-oid dehydrogenase 3 activities in human and rat testes[J]. Chemico-Biological Interactions, 195(3): 180-188. [41] Zhang M, Wei H, Liu T, et al. 2020. Potential GnRH and ste-roidogenesis pathways in the scallop Patinopecten yes-soensis[J]. Journal of Steroid Biochemistry and Molecu-lar Biology, 204: 105756 [42] Zhai H N, Zhou J, Cai Z H, et al. 2012. Cloning, characteriza-tion, and expression analysis of a putative 17beta-hy-droxysteroid dehy-drogenase 11 in the abalone, Haliotis diversicolor supertexta[J]. Journal of Steroid Biochemis-try, 130(1/2): 57-63. [43] Zhao Y, Chen K, Liu F, et al. 2022. Dynamic gene expression and alternative splicing events demonstrate co-regula-tion of testicular differentiation and maturation by the brain and gonad in common carp[J]. Frontiers in Endo-crinology, 12: 820463. [44] Zhou L, Li M, Wang D. 2021. Role of sex steroids in fish sex determination and differentiation as revealed by gene ed-iting[J]. General and Comparative Endocrinology, 313: 113893. [45] Zirkin B R,Papadopoulos V. 2018. Leydig cells: Formation, function, and regulation[J]. Biology of Reproduction, 99(1): 101-111. |
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