Histological and sex-determining genes expression effects of 17α-methyltestosterone on mandarin fish Siniperca chuatsi gonad development

Background 17α-methyltestosterone (MT) as a synthetic androgen has been widely used in sex reversal of fish research, but there was no published report on sex reversal of mandarin fish by MT. Moreover, comparative transcriptome analysis of testis and ovarian tissue is still lacking in mandarin fish. We used histological analyses together with RNA sequencing to characterize mandarin fish gonadal transcriptomes and investigate the effects of MT on the sex ratio, survival rate, growth, gonadal differentiation.ResultsMandarin fish treated with dietary MT at 50 mg/kg and 100 mg/kg dosages were successfully induced to all-male stock (male rate 100%), compared with the control group (51.11%). The survival rate of fish in the MT treated and control groups were not significant different. MT were significantly inhibited the growth of the MT treatment group ( P < 0.05) at the 20 dph-120 dph, however, the weight and length in the MT treated and control groups were not significant different at the 180 dph and 240 dph. MT treatment promoted the development of testis, but inhibited the gonadosomatic index (GSI) and the levels of serum steroid hormone (T and E 2 ). This work screened out the genes related to the sex determination and differentiation of the fish by sequencing and analysis of the transcriptome of the ovary and testis. The masculinization of mandarin fish was also demonstrated by the expression patterns of sex-specific genes, dmrt1, sox9, foxl2 and cyp19a1a : the gonads of MT-treated fish exclusively expressed male-specific dmrt1 and sox9 with no expression of female-specific foxl2 and cyp19a1a .Conclusion This study suggests that 17α-methyltestosterone successfully induced all-male stock and we select the part of the genes ( dmrt1, sox9, foxl2 and cyp19a1a ) related to sex determination and differentiation.

[1]  Yong Zhang,et al.  Expression profiles of dmrts and foxls during gonadal development and sex reversal induced by 17α-methyltestosterone in the orange-spotted grouper. , 2019, General and comparative endocrinology.

[2]  Hongwei Yan,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 , 2018, Fish Physiology and Biochemistry.

[3]  T. Kocher,et al.  Transcriptome display during tilapia sex determination and differentiation as revealed by RNA-Seq analysis , 2018, BMC Genomics.

[4]  Guan-Chung Wu,et al.  Gonadal development and expression of sex-specific genes during sex differentiation in the Japanese eel. , 2017, General and comparative endocrinology.

[5]  Haitham G. Abo‐Al‐Ela.  Hormones and fish monosex farming: A spotlight on immunity. , 2018, Fish & shellfish immunology.

[6]  Xiumei Liu,et al.  Comparative transcriptome analysis of ovary and testis reveals potential sex-related genes and pathways in spotted knifejaw Oplegnathus punctatus. , 2017, Gene.

[7]  J. A. Luckenbach,et al.  Establishment of monosex female production of sablefish (Anoplopoma fimbria) through direct and indirect sex control , 2017 .

[8]  Hongxia Hu,et al.  Dimorphic expression of sex-related genes in different gonadal development stages of sterlet, Acipenser ruthenus, a primitive fish species , 2017, Fish Physiology and Biochemistry.

[9]  W. Tao,et al.  CRISPR/Cas9-induced disruption of wt1a and wt1b reveals their different roles in kidney and gonad development in Nile tilapia. , 2017, Developmental biology.

[10]  N. Gemmell,et al.  Histological and transcriptomic effects of 17α-methyltestosterone on zebrafish gonad development , 2017, BMC Genomics.

[11]  S. Fatima,et al.  Sex reversal of brook trout (Salvelinus fontinalis) by 17α-methyltestosterone exposure: A serial experimental approach to determine optimal timing and delivery regimes. , 2016, Animal reproduction science.

[12]  Tohru Kobayashi,et al.  Androgen induces gonadal soma-derived factor, Gsdf, in XX gonads correlated to sex-reversal but not Dmrt1 directly, in the teleost fish, northern medaka (Oryzias sakaizumii) , 2016, Molecular and Cellular Endocrinology.

[13]  Hongwei Liang,et al.  Molecular mechanism of endocrine system impairment by 17α-methyltestosterone in gynogenic Pengze crucian carp offspring. , 2016, Ecotoxicology and environmental safety.

[14]  Chao Yang,et al.  Genome-Wide Identification and Transcriptome-Based Expression Profiling of the Sox Gene Family in the Nile Tilapia (Oreochromis niloticus) , 2016, International journal of molecular sciences.

[15]  Han-Ping Wang,et al.  Effects of 17α-Methyltestosterone and Aromatase Inhibitor Letrozole on Sex Reversal, Gonadal Structure, and Growth in Yellow Catfish Pelteobagrus fulvidraco , 2015, The Biological Bulletin.

[16]  Lina Sun,et al.  Antagonistic roles of Dmrt1 and Foxl2 in sex differentiation via estrogen production in tilapia as demonstrated by TALENs. , 2013, Endocrinology.

[17]  Linyan Zhou,et al.  Characterization of Gonadal Transcriptomes from Nile Tilapia (Oreochromis niloticus) Reveals Differentially Expressed Genes , 2013, PloS one.

[18]  Hua Tian,et al.  Genetic Manipulation of Sex Ratio for the Large-Scale Breeding of YY Super-Male and XY All-Male Yellow Catfish (Pelteobagrus fulvidraco (Richardson)) , 2012, Marine Biotechnology.

[19]  Sun Zhong-zhi.  Annual maturation of testis correlated with sex steroids level in male barfin flounder broodstocks , 2012 .

[20]  Shaojun Liu,et al.  Massive Production of All-female Diploids and Triploids in the Crucian Carp , 2011, International journal of biological sciences.

[21]  R. Phelps,et al.  A non-paradoxical dose response to 17α-methyltestosterone by Nile tilapia Oreochromis niloticus (L.): effects on the sex ratio, growth and gonadal development , 2011 .

[22]  G. Taranger,et al.  The production of hermaphrodites of Atlantic cod (Gadus morhua) by masculinization with orally administered 17-α-methyltestosterone, and subsequent production of all-female cod populations. , 2011 .

[23]  J. Osborne,et al.  Effects of oestradiol-17β or 17α-methyltestosterone administration on gonadal differentiation of largemouth bass Micropterus salmoides (Lacepède) , 2009 .

[24]  M. Maliqueo,et al.  Testosterone-induced downregulation of anti-Müllerian hormone expression in granulosa cells from small bovine follicles , 2009, Endocrine.

[25]  Y. Guiguen,et al.  Successful production of monosex female brook trout Salvelinus fontinalis using gynogenetic sex reversed males by a combination of methyltestosterone immersion and oral treatments. , 2009 .

[26]  J. Godwin,et al.  Sex determination in flatfishes: Mechanisms and environmental influences. , 2009, Seminars in cell & developmental biology.

[27]  M. Gerstein,et al.  RNA-Seq: a revolutionary tool for transcriptomics , 2009, Nature Reviews Genetics.

[28]  F. Piferrer,et al.  Sex-related changes in estrogen receptors and aromatase gene expression and enzymatic activity during early development and sex differentiation in the European sea bass (Dicentrarchus labrax). , 2008, General and comparative endocrinology.

[29]  B. Williams,et al.  Mapping and quantifying mammalian transcriptomes by RNA-Seq , 2008, Nature Methods.

[30]  I. Goodhead,et al.  Dynamic repertoire of a eukaryotic transcriptome surveyed at single-nucleotide resolution , 2008, Nature.

[31]  R. Horiguchi,et al.  Molecular cloning and quantitative expression of sexually dimorphic markers Dmrt1 and Foxl2 during female-to-male sex change in Epinephelus merra. , 2008, General and comparative endocrinology.

[32]  Tohru Kobayashi,et al.  Sexual Dimorphic Expression of Genes in Gonads During Early Differentiation of a Teleost Fish, the Nile Tilapia Oreochromis niloticus1 , 2008, Biology of reproduction.

[33]  H. Rashid,et al.  Fugu (Takifugu rubripes) Sexual Differentiation: CYP19 Regulation and Aromatase Inhibitor Induced Testicular Development , 2007, Sexual Development.

[34]  B. Senthilkumaran,et al.  Ontogenic expression patterns of several nuclear receptors and cytochrome P450 aromatases in brain and gonads of the Nile tilapia Oreochromis niloticus suggests their involvement in sex differentiation , 2005, Fish Physiology and Biochemistry.

[35]  M. Schartl,et al.  Divergent expression patterns of Sox9 duplicates in teleosts indicate a lineage specific subfunctionalization , 2005, Development Genes and Evolution.

[36]  R. Veitia,et al.  An evolutionary and functional analysis of FoxL2 in rainbow trout gonad differentiation. , 2004, Journal of molecular endocrinology.

[37]  N. Woo,et al.  Influence of oral administration of estradiol-17β and testosterone on growth, digestion, food conversion and metabolism in the underyearling red sea bream, Chrysophrys major , 1993, Fish Physiology and Biochemistry.

[38]  M. Schartl,et al.  Absence of the Candidate Male Sex-Determining Gene dmrt1b(Y) of Medaka from Other Fish Species , 2003, Current Biology.

[39]  P. Carlsson,et al.  Forkhead transcription factors: key players in development and metabolism. , 2002, Developmental biology.

[40]  P. Kestemont,et al.  Sex control and sex determinism study in Eurasian perch, Perca fluviatilis, by use of hormonally sex-reversed male breeders , 2002 .

[41]  F. Weltzien,et al.  Spermatogenesis and related plasma androgen levels in Atlantic halibut (Hippoglossus hippoglossus L.). , 2002, Comparative biochemistry and physiology. Part A, Molecular & integrative physiology.

[42]  R. Devlin,et al.  Sex determination and sex differentiation in fish: an overview of genetic, physiological, and environmental influences , 2002 .

[43]  Shuichi Asakawa,et al.  DMY is a Y-specific DM-domain gene required for male development in the medaka fish , 2002, Nature.

[44]  A. Place,et al.  Developmental expression of cytochrome P450 aromatase genes (CYP19a and CYP19b) in zebrafish fry (Danio rerio). , 2001, The Journal of experimental zoology.

[45]  Y. Guiguen,et al.  Endocrine and environmental aspects of sex differentiation in fish , 1999, Cellular and Molecular Life Sciences CMLS.

[46]  P. Donahoe,et al.  Müllerian-inhibiting substance regulates androgen synthesis at the transcriptional level. , 1999, Endocrinology.

[47]  V. Rouiller-Fabre,et al.  Effect of anti-Mullerian hormone on Sertoli and Leydig cell functions in fetal and immature rats. , 1998, Endocrinology.

[48]  S. Yamashita,et al.  Rainbow trout SOX9: cDNA cloning, gene structure and expression. , 1997, Gene.

[49]  A. Sinclair,et al.  Mutations in SRY and SOX9: Testis‐determining genes , 1997, Human mutation.

[50]  T. Pandian,et al.  Hormonal induction of sex reversal and progeny testing in the zebra cichlid Cichlasoma nigrofasciatum , 1996 .

[51]  K. Yamauchi,et al.  Ovarian development and serum sex steroid and vitellogenin profiles in the female cultured sturgeon hybrid, the bester , 1996 .

[52]  K. Yamauchi,et al.  Testicular development and serum sex steroid profiles during the annual sexual cycle of the male sturgeon hybrid, the bester , 1996 .

[53]  T. Pandian,et al.  Production of ZZ females in the female-heterogametic black molly, Poecilia sphenops, by endocrine sex reversal and progeny testing , 1995 .

[54]  F. Piferrer,et al.  Development of sex control techniques for European sea bass (Dicentrarchus labrax L.) aquaculture: effects of dietary 17 α-methyltestosterone prior to sex differentiation , 1995 .

[55]  Y. Ito,et al.  Aromatase cytochrome P450, the enzyme responsible for estrogen biosynthesis. , 1994, Endocrine reviews.

[56]  G. Mair,et al.  Conditions for effective androgen sex reversal in Oreochromis niloticus (L.) , 1994 .

[57]  B. A. Simco,et al.  Hormonal sex manipulation and evidence for female homogamety in channel catfish. , 1990, General and comparative endocrinology.

[58]  W. Shelton,et al.  Induction of gynogenesis and sex reversal in silver carp , 1988 .

[59]  A. J. Matty,et al.  The effect of feeding methyltestosterone on the growth and body composition of common carp (Cyprinus carpio L.). , 1980, General and comparative endocrinology.

[60]  H. Clemens,et al.  The Production of Unisexual Broods by Tilapia mossambica Sex‐reversed with Methyl Testosterone , 1968 .