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| Expression Analysis of Genes Related to Sex Differentiation in Tiger Puffer Takifugu rubripes Juveniles |
| JIANG Jieming1,2, SHEN Xufang2,3, LIU Ying1,2,4, ZHANG Qi1, ZHOU Huiting1, WANG Jia1, YAN Hongwei1,2, WANG Chenqi1, LIU Qi1,2 |
1. Dalian Ocean University, Dalian 116023, China;
2. Key Laboratory of Environment Controlled Aquaculture, Ministry of Education, Dalian 116023, China;
3. Liaoning Normal University, Dalian 116029, China;
4. Pilot National Laboratory for Marine Science and Technology, Qingdao 266237, China |
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Abstract In order to clarify the expression patterns of sex differentiation-related genes during the critical period of sex differentiation in tiger puffer Takifugu rubripes, the histological characteristics of the gonads were observed in tiger puffer juveniles at 40, 60 and 80 d post hatching firstly. Then, after the genetic sex verification of individual, the expression patterns of amhr2, dmrt1, gsdf, cyp19a1a, foxl2 and dnmt1 genes were determined in the gonads of XX and XY tiger puffer juveniles by real-time quantitative PCR (qPCR). The results showed that the expression of the above genes showed sexual dimorphism obviously, without significant difference in the expression level of amhr2 gene in the gonads of XX and XY tiger puffer juveniles at 40 and 60 d post hatching (P>0.05), significantly higher expression level of amhr2 gene in the gonads of XX juveniles than that in XY juveniles at 80 d post hatching (P<0.05). The expression levels of dmrt1 and gsdf genes in the gonads of XY juveniles were increased significantly from 20 d, and the expression levels in the gonads of XY juveniles were significantly higher than that in XX juveniles at all the three sampling points (P<0.05). The expression levels of cyp19a1a and foxl2 genes in the gonads of XX juveniles were significantly higher than that in XY juveniles at the 40, 60 and 80 d post hatching (P<0.05). There was no significant difference in the expression level of dnmt1 gene in the gonads of XX and XY tiger puffer juveniles at 40 and 60 d post hatching (P>0.05), but the expression level of dnmt1 gene in the gonads of XY tiger puffer juveniles was significantly higher than that in XX juveniles at 80 d post hatching (P<0.05). The findings demonstrated that dmrt1 and gsdf genes played critical roles in testicular differentiation, while cyp19a1a and foxl2 genes played critical roles in ovarian differentiation in tiger puffer juveniles. The findings laid a foundation for functional analysis of the sex differentiation-related genes during the development of sex differentiation of tiger puffer.
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Received: 22 March 2021
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[1] 杨旭,吴东舒,华昕彤,等.红鳍东方鲀3个白介素基因的序列特征及表达分析[J].水产科学,2021,40(1):64-71.
[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] 马爱军,房金岑,陈蓝荪,等.河豚鱼产业调研报告(上)[J].海洋与渔业·水产前沿,2014(7):87-92.
[4] KIKUCHI K, KAI W, HOSOKAWA A, et al. The sex-determining locus in the tiger pufferfish, Takifugu rubripes[J].Genetics,2007,175(4):2039-2042.
[5] APARICIO S, CHAPMAN J, STUPKA E, et al. Whole-genome shotgun assembly and analysis of the genome of Fugu rubripes[J].Science,2002,297(5585):1301-1310.
[6] KAMIYA T, KAI W, TASUMI S, et al. A trans-species missense SNP in Amhr2 is associated with sex determination in the tiger pufferfish, Takifugu rubripes (fugu)[J].PLoS Genetics,2012,8(7):e1002798.
[7] MATSUNAGA T, IEDA R, HOSOYA S, et al. An efficient molecular technique for sexing tiger pufferfish (fugu) and the occurrence of sex reversal in a hatchery population[J].Fisheries Science,2014,80(5):933-942.
[8] RASHID H, KITANO H, LEE K H, et al. Fugu (Takifugu rubripes) sexual differentiation:CYP19 regulation and aromatase inhibitor induced testicular development[J].Sexual Development,2007,1(5):311-322.
[9] YAMAGUCHI A, LEE K H, FUJIMOTO H, et al. Expression of the DMRT gene and its roles in early gonadal development of the Japanese pufferfish Takifugu rubripes[J].Comparative Biochemistry and Physiology Part D:Genomics and Proteomics,2006,1(1):59-68.
[10] YAN H W, SHEN X F, CUI X, et al. Identification of genes involved in gonadal sex differentiation and the dimorphic expression pattern in Takifugu rubripes gonad at the early stage of sex differentiation[J].Fish Physiology and Biochemistry,2018,44(5):1275-1290.
[11] 张旭,唐瞻杨,黄凯,等.维生素E对尼罗罗非鱼生长和性腺发育影响的研究[J].水产科学,2020,39(6):887-893.
[12] 孙红娟,陈仲,贺迪,等.仿刺参SARM基因的克隆、表达和功能研究[J].水产科学,2021,40(1):1-9.
[13] 梅洁,桂建芳.鱼类性别异形和性别决定的遗传基础及其生物技术操控[J].中国科学:生命科学,2014,44(12):1198-1212.
[14] PFENNIG F, STANDKE A, GUTZEIT H O. The role of Amh signaling in teleost fish—multiple functions not restricted to the gonads[J].General and Comparative Endocrinology,2015,223:87-107.
[15] HATTORI R S, MURAI Y, OURA M, et al. A Y-linked anti-Müllerian hormone duplication takes over a critical role in sex determination[J].Proceedings of the National Academy of Sciences of the United States of America,2012,109(8):2955-2959.
[16] LI M H, SUN Y L, ZHAO J E, et al. A tandem duplicate of anti-Müllerian hormone with a missense SNP on the Y chromosome is essential for male sex determination in Nile tilapia, Oreochromis niloticus[J].PLoS Genetics,2015,11(11):e1005678.
[17] MORINAGA C, SAITO D, NAKAMURA S, et al. The hotei mutation of medaka in the anti-Müllerian hormone receptor causes the dysregulation of germ cell and sexual development[J].Proceedings of the National Academy of Sciences of the United States of America,2007,104(23):9691-9696.
[18] MULLEN R D, ONTIVEROS A E, MOSES M M, et al. AMH and AMHR2 mutations:a spectrum of reproductive phenotypes across vertebrate species[J].Developmental Biology,2019,455(1):1-9.
[19] HERPIN A, SCHARTL M. Dmrt1 genes at the crossroads:a widespread and central class of sexual development factors in fish[J].The FEBS Journal,2011,278(7):1010-1019.
[20] PICARD M A L, COSSEAU C, MOUAHID G, et al. The roles of Dmrt (Double sex/Male-abnormal-3 Related transcription factor) genes in sex determination and differentiation mechanisms:ubiquity and diversity across the animal kingdom[J].Comptes Rendus Biologies,2015,338(7):451-462.
[21] MASUYAMA H, YAMADA M, KAMEI Y, et al. Dmrt1 mutation causes a male-to-female sex reversal after the sex determination by Dmy in the medaka[J].Chromosome Research,2012,20(1):163-176.
[22] LIN Q H, MEI J, LI Z, et al. Distinct and cooperative roles of amh and dmrt1 in self-renewal and differentiation of male germ cells in zebrafish[J].Genetics,2017,207(3):1007-1022.
[23] SAWATARI E, SHIKINA S, TAKEUCHI T, et al. A novel transforming growth factor-beta superfamily member expressed in gonadal somatic cells enhances primordial germ cell and spermatogonial proliferation in rainbow trout (Oncorhynchus mykiss)[J].Developmental Biology,2007,301(1):266-275.
[24] MYOSHO T, OTAKE H, MASUYAMA H, et al. Tracing the emergence of a novel sex-determining gene in medaka, Oryzias luzonensis[J].Genetics,2012,191(1):163-170.
[25] IMAI T, SAINO K, MATSUDA M. Mutation of gonadal soma-derived factor induces medaka XY gonads to undergo ovarian development[J].Biochemical and Biophysical Research Communications,2015,467(1):109-114.
[26] ZHANG X, GUAN G J, LI M Y, et al. Autosomal gsdf acts as a male sex initiator in the fish medaka[J].Scientific Reports,2016,6:19738.
[27] JIANG D N, YANG H H, LI M H, et al. Gsdf is a downstream gene of dmrt1 that functions in the male sex determination pathway of the Nile tilapia[J].Molecular Reproduction and Development,2016,83(6):497-508.
[28] 高莹莹,刘新富,胡鹏,等.暗纹东方鲀芳香化酶基因的结构及表达分析[J].水产科学,2020,39(1):30-39.
[29] LAU ESTHERS W, ZHANG Z W, QIN M M, et al. Knockout of zebrafish ovarian aromatase gene (cyp19a1a) by TALEN and CRISPR/Cas9 leads to all-male offspring due to failed ovarian differentiation[J].Scientific Reports,2016,6:37357.
[30] WU K, SONG W Y, ZHANG Z W, et al. Disruption of dmrt1 rescues the all-male phenotype of the cyp19a1a mutant in zebrafish—a novel insight into the roles of aromatase/estrogens in gonadal differentiation and early folliculogenesis[J].Development,2020,147(4):dev182758.
[31] BOULANGER L, PANNETIER M, GALL L, et al. FOXL2 is a female sex-determining gene in the goat[J].Current Biology,2014,24(4):404-408.
[32] ZHANG X B, LI M R, MA H, et al. Mutation of foxl2 or cyp19a1a results in female to male sex reversal in XX Nile tilapia[J].Endocrinology,2017,158(8):2634-2647.
[33] YANG Y J, WANG Y, LI Z, et al. Sequential, divergent, and cooperative requirements of Foxl2a and Foxl2b in ovary development and maintenance of zebrafish[J].Genetics,2017,205(4):1551-1572.
[34] LIU H F, MU X J, GUI L, et al. Characterization and gonadal expression of FOXL2 relative to Cyp19a genes in spotted scat Scatophagus argus[J].Gene,2015,561(1):6-14. |
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