Feminization of channel catfish with 17β-oestradiol involves methylation and expression of a specific set of genes independent of the sex determination region

ABSTRACT Exogenous oestrogen 17β-oestradiol (E2) has been shown to effectively induce feminization in teleosts. However, the molecular mechanisms underlying the process remain unclear. Here, we determined global DNA methylation and gene expression profiles of channel catfish (Ictalurus punctatus) during early sex differentiation after E2 treatment. Overall, the levels of global DNA methylation after E2 treatment were not significantly different from those of controls. However, a specific set of genes were differentially methylated, which included many sex differentiation-related pathways, such as MARK signalling, adrenergic signalling, Wnt signalling, GnRH signalling, ErbB signalling, and ECM–receptor interactions. Many genes involved in these pathways were also differentially expressed after E2 treatment. Specifically, E2 treatments resulted in upregulation of female-related genes and downregulation of male-related genes in genetic males during sex reversal. However, E2-induced sex reversal did not cause sex-specific changes in methylation profiles or gene expression within the sex determination region (SDR) on chromosome 4, suggesting that E2-induced sex reversal was a downstream process independent of the sex determination process that was regulated by sex-specific methylation within the SDR.

[1]  Zhanjiang Liu,et al.  Identification of an Epigenetically Marked Locus within the Sex Determination Region of Channel Catfish , 2022, International journal of molecular sciences.

[2]  D. Jerry,et al.  Induction of precocious females in the protandrous barramundi (Lates calcarifer) through implants containing 17β-estradiol - effects on gonadal morphology, gene expression and DNA methylation of key sex genes , 2021, Aquaculture.

[3]  I. Ábrahám,et al.  Estradiol-Induced Epigenetically Mediated Mechanisms and Regulation of Gene Expression , 2020, International journal of molecular sciences.

[4]  Yong Lin,et al.  Effects of gonadotropin-releasing hormone analog (GnRHa) immunization on the gonadal transcriptome and proteome of tilapia (Oreochromis niloticus). , 2020, Comparative biochemistry and physiology. Part D, Genomics & proteomics.

[5]  N. Gemmell,et al.  Stress, novel sex genes, and epigenetic reprogramming orchestrate socially controlled sex change , 2018, Science Advances.

[6]  Lina Sun,et al.  Roles of estrogens in fish sexual plasticity and sex differentiation. , 2019, General and comparative endocrinology.

[7]  Jiaqiang Xiong,et al.  Proteomic analysis of mouse ovaries during the prepubertal stages , 2019, Experimental cell research.

[8]  S. Meng,et al.  Testicular transcriptome alterations in zebrafish (Danio rerio) exposure to 17β-estradiol. , 2019, Chemosphere.

[9]  Shikai Liu,et al.  The Y chromosome sequence of the channel catfish suggests novel sex determination mechanisms in teleost fish , 2019, BMC Biology.

[10]  B. Draper,et al.  Genetic regulation of sex determination and maintenance in zebrafish (Danio rerio). , 2019, Current topics in developmental biology.

[11]  S. Munck,et al.  A Label-free Multicolor Optical Surface Tomography (ALMOST) imaging method for nontransparent 3D samples , 2019, BMC Biology.

[12]  Zhiwei Zhang,et al.  Roles of Figla/figla in Juvenile Ovary Development and Follicle Formation During Zebrafish Gonadogenesis. , 2018, Endocrinology.

[13]  Arun S. Seetharam,et al.  Insights into teleost sex determination from the Seriola dorsalis genome assembly , 2018, BMC Genomics.

[14]  S. Ru,et al.  Estrogenic effects associated with bisphenol a exposure in male zebrafish (Danio rerio) is associated with changes of endogenous 17β-estradiol and gene specific DNA methylation levels. , 2017, General and comparative endocrinology.

[15]  B. Capel Vertebrate sex determination: evolutionary plasticity of a fundamental switch , 2017, Nature Reviews Genetics.

[16]  N. Zhang,et al.  Utilization of a Wheat660K SNP array-derived high-density genetic map for high-resolution mapping of a major QTL for kernel number , 2017, Scientific Reports.

[17]  Songlin Chen,et al.  Genome editing reveals dmrt1 as an essential male sex-determining gene in Chinese tongue sole (Cynoglossus semilaevis) , 2017, Scientific Reports.

[18]  L. J. Wang,et al.  Transcriptome analysis of the uniparous and multiparous goats ovaries. , 2016, Reproduction in domestic animals = Zuchthygiene.

[19]  Lixue Sun,et al.  Global DNA Methylation Changes in Nile Tilapia Gonads during High Temperature-Induced Masculinization , 2016, PloS one.

[20]  S. Koren,et al.  The channel catfish genome sequence provides insights into the evolution of scale formation in teleosts , 2016, Nature Communications.

[21]  D. Lan,et al.  Toward Understanding the Genetic Basis of Yak Ovary Reproduction: A Characterization and Comparative Analyses of Estrus Ovary Transcriptiome in Yak and Cattle , 2016, PloS one.

[22]  Xin Hu,et al.  Twist2 promotes kidney cancer cell proliferation and invasion by regulating ITGA6 and CD44 expression in the ECM-receptor interaction pathway , 2016, OncoTargets and therapy.

[23]  O. Aral,et al.  The effects of different proportions of the 17β-estradiol and 17α-methyltestosterone on growth, sex reversal and skin colouration of the electric blue hap (Sciaenochromis ahli Trewavas, 1935) , 2016 .

[24]  Jean-Philippe Vert,et al.  Changes in correlation between promoter methylation and gene expression in cancer , 2015, BMC Genomics.

[25]  F. Brion,et al.  Localization of steroidogenic enzymes and Foxl2a in the gonads of mature zebrafish (Danio rerio). , 2015, Comparative biochemistry and physiology. Part A, Molecular & integrative physiology.

[26]  X. Mu,et al.  Characterization and gonadal expression of FOXL2 relative to Cyp19a genes in spotted scat Scatophagus argus. , 2015, Gene.

[27]  Steven L Salzberg,et al.  HISAT: a fast spliced aligner with low memory requirements , 2015, Nature Methods.

[28]  Zhanjiang Liu,et al.  Construction of a high-density, high-resolution genetic map and its integration with BAC-based physical map in channel catfish , 2014, DNA research : an international journal for rapid publication of reports on genes and genomes.

[29]  W. Huber,et al.  Moderated estimation of fold change and dispersion for RNA-seq data with DESeq2 , 2014, Genome Biology.

[30]  Paul Theodor Pyl,et al.  HTSeq—a Python framework to work with high-throughput sequencing data , 2014, bioRxiv.

[31]  Björn Usadel,et al.  Trimmomatic: a flexible trimmer for Illumina sequence data , 2014, Bioinform..

[32]  H. Ostrer,et al.  Mutations in MAP3K1 tilt the balance from SOX9/FGF9 to WNT/β-catenin signaling. , 2014, Human molecular genetics.

[33]  Mathieu Blanchette,et al.  The relationship between DNA methylation, genetic and expression inter-individual variation in untransformed human fibroblasts , 2014, Genome Biology.

[34]  W. Dębek,et al.  Serum AMH in Physiology and Pathology of Male Gonads , 2013, International journal of endocrinology.

[35]  M. Megias,et al.  Environmental induced methylation changes associated with seawater adaptation in brown trout , 2013 .

[36]  F. Piferrer Epigenetics of sex determination and gonadogenesis , 2013, Developmental dynamics : an official publication of the American Association of Anatomists.

[37]  S. Wells,et al.  Gadd45γ and Map3k4 Interactions Regulate Mouse Testis Determination via p38 MAPK-Mediated Control of Sry Expression , 2012, Developmental cell.

[38]  C. Niehrs,et al.  GADD45G functions in male sex determination by promoting p38 signaling and Sry expression. , 2012, Developmental cell.

[39]  T. Benfey,et al.  Hormonal sex reversal in Atlantic cod, Gadus morhua , 2012 .

[40]  Francine E. Garrett-Bakelman,et al.  methylKit: a comprehensive R package for the analysis of genome-wide DNA methylation profiles , 2012, Genome Biology.

[41]  Masaru Nakamura,et al.  Diversity and Plasticity of Sex Determination and Differentiation in Fishes , 2012, Sexual Development.

[42]  E. Peatman,et al.  Identification of a sex-linked marker for channel catfish. , 2012, Animal genetics.

[43]  S. Munger,et al.  Sex and the circuitry: progress toward a systems‐level understanding of vertebrate sex determination , 2012, Wiley interdisciplinary reviews. Systems biology and medicine.

[44]  Guangchuang Yu,et al.  clusterProfiler: an R package for comparing biological themes among gene clusters. , 2012, Omics : a journal of integrative biology.

[45]  Steven L Salzberg,et al.  Fast gapped-read alignment with Bowtie 2 , 2012, Nature Methods.

[46]  P. Manolakou,et al.  Sex determination strategies in 2012: towards a common regulatory model? , 2012, Reproductive Biology and Endocrinology.

[47]  D. Zarkower,et al.  Sex and the singular DM domain: insights into sexual regulation, evolution and plasticity , 2012, Nature Reviews Genetics.

[48]  P. Sridevi,et al.  FTZ-F1 and FOXL2 up-regulate catfish brain aromatase gene transcription by specific binding to the promoter motifs. , 2012, Biochimica et biophysica acta.

[49]  P. Sridevi,et al.  Cloning and differential expression of FOXL2 during ovarian development and recrudescence of the catfish, Clarias gariepinus. , 2011, General and comparative endocrinology.

[50]  L. Ribas,et al.  DNA Methylation of the Gonadal Aromatase (cyp19a) Promoter Is Involved in Temperature-Dependent Sex Ratio Shifts in the European Sea Bass , 2011, PLoS genetics.

[51]  Felix Krueger,et al.  Bismark: a flexible aligner and methylation caller for Bisulfite-Seq applications , 2011, Bioinform..

[52]  H. Wickham ggplot2 , 2011 .

[53]  H. Ostrer,et al.  Mutations in MAP3K1 cause 46,XY disorders of sex development and implicate a common signal transduction pathway in human testis determination. , 2010, American journal of human genetics.

[54]  B. Brunström,et al.  DNA methylation levels in the 5' flanking region of the vitellogenin I gene in liver and brain of adult zebrafish (Danio rerio)--sex and tissue differences and effects of 17alpha-ethinylestradiol exposure. , 2010, Aquatic toxicology.

[55]  A. Navratil,et al.  GnRH signaling, the gonadotrope and endocrine control of fertility , 2010, Frontiers in Neuroendocrinology.

[56]  I. Simon,et al.  DNA methylation and gene expression , 2010, Wiley interdisciplinary reviews. Systems biology and medicine.

[57]  A. Sinclair,et al.  The avian Z-linked gene DMRT1 is required for male sex determination in the chicken , 2009, Nature.

[58]  H. Ostrer,et al.  Loss of Mitogen-Activated Protein Kinase Kinase Kinase 4 (MAP3K4) Reveals a Requirement for MAPK Signalling in Mouse Sex Determination , 2009, PLoS biology.

[59]  Gonçalo R. Abecasis,et al.  The Sequence Alignment/Map format and SAMtools , 2009, Bioinform..

[60]  J. I. Fernandino,et al.  Dimorphic Expression of dmrt1 and cyp19a1 (Ovarian Aromatase) during Early Gonadal Development in Pejerrey, Odontesthes bonariensis , 2009, Sexual Development.

[61]  Masaru Nakamura,et al.  Dual Roles of cyp19a1a in Gonadal Sex Differentiation and Development in the Protandrous Black Porgy, Acanthopagrus schlegeli1 , 2008, Biology of reproduction.

[62]  I. Katsiadaki,et al.  Global genomic methylation levels in the liver and gonads of the three-spine stickleback (Gasterosteus aculeatus) after exposure to hexabromocyclododecane and 17-beta oestradiol. , 2008, Environment international.

[63]  C. Tyler,et al.  Estrogen-induced alterations in amh and dmrt1 expression signal for disruption in male sexual development in the zebrafish. , 2007, Environmental science & technology.

[64]  Tohru Kobayashi,et al.  Foxl2 up-regulates aromatase gene transcription in a female-specific manner by binding to the promoter as well as interacting with ad4 binding protein/steroidogenic factor 1. , 2007, Molecular endocrinology.

[65]  B. Sinervo Sex Determination and Differentiation , 2007 .

[66]  S. Levy,et al.  Ovarian gene expression in the absence of FIGLA, an oocyte-specific transcription factor , 2007, BMC Developmental Biology.

[67]  M. Ferguson-Smith The Evolution of Sex Chromosomes and Sex Determination in Vertebrates and the Key Role of DMRT1 , 2006, Sexual Development.

[68]  P. Manolakou,et al.  Molecular patterns of sex determination in the animal kingdom: a comparative study of the biology of reproduction , 2006, Reproductive biology and endocrinology : RB&E.

[69]  Kunio Kobayashi,et al.  Comparison of response to 17β‐estradiol and 17β‐trenbolone among three small fish species , 2006 .

[70]  R. Veitia,et al.  FOXL2 activates P450 aromatase gene transcription: towards a better characterization of the early steps of mammalian ovarian development. , 2006, Journal of molecular endocrinology.

[71]  Y. Nagahama,et al.  Molecular cloning and analysis of gonadal expression of Foxl2 in the medaka, Oryzias latipes. , 2006, Biochemical and biophysical research communications.

[72]  T. Wibbels,et al.  Dmrt1 expression in response to estrogen treatment in a reptile with temperature-dependent sex determination. , 2006, Journal of experimental zoology. Part B, Molecular and developmental evolution.

[73]  D. Schlessinger,et al.  Foxl2 is required for commitment to ovary differentiation. , 2005, Human molecular genetics.

[74]  Masaru Nakamura,et al.  Evidence that estrogen regulates the sex change of honeycomb grouper (Epinephelus merra), a protogynous hermaphrodite fish. , 2005, Journal of experimental zoology. Part A, Comparative experimental biology.

[75]  S. Ojeda,et al.  erbB-1 and erbB-4 receptors act in concert to facilitate female sexual development and mature reproductive function. , 2005, Endocrinology.

[76]  H. Yao The pathway to femaleness: current knowledge on embryonic development of the ovary , 2005, Molecular and Cellular Endocrinology.

[77]  Shoko Imai,et al.  Effects of 17beta-estradiol on the reproduction of Java-medaka (Oryzias javanicus), a new test fish species. , 2005, Marine pollution bulletin.

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

[79]  J. Brillard,et al.  Isolation of chicken homolog of the FOXL2 gene and comparison of its expression patterns with those of aromatase during ovarian development , 2004, Developmental dynamics : an official publication of the American Association of Anatomists.

[80]  Yosef Yarden,et al.  Signal transduction and oncogenesis by ErbB/HER receptors. , 2004, International journal of radiation oncology, biology, physics.

[81]  Masaru Nakamura,et al.  The role estrogens play in sex differentiation and sex changes of fish , 2003, Fish Physiology and Biochemistry.

[82]  T. Pandian,et al.  Recent Advances in Hormonal Induction of Sex-Reversal in Fish , 2003 .

[83]  N. Rosenthal,et al.  Normal Female Sexual Development Requires Neuregulin–erbB Receptor Signaling in Hypothalamic Astrocytes , 2003, The Journal of Neuroscience.

[84]  M. James,et al.  Intestinal and hepatic microsomal metabolism of testosterone and progesterone by a 3α-hydroxysteroid dehydrogenase to the 3α-hydroxy derivatives in the channel catfish, Ictalurus punctatus , 2002, The Journal of Steroid Biochemistry and Molecular Biology.

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

[86]  R. Seger,et al.  Intracellular signaling pathways mediated by the gonadotropin-releasing hormone (GnRH) receptor. , 2001, Archives of medical research.

[87]  Francesc Piferrer,et al.  Endocrine sex control strategies for the feminization of teleost fish , 2001 .

[88]  A. Bernot,et al.  DMRT1 expression during gonadal differentiation and spermatogenesis in the rainbow trout, Oncorhynchus mykiss. , 2000, Biochimica et biophysica acta.

[89]  M. Hiroi,et al.  Sandwich configuration of type I collagen suppresses progesterone production in primary cultured porcine granulosa cells by reducing gene expression of cytochrome P450 cholesterol side-chain cleavage enzyme. , 2000, Archives of biochemistry and biophysics.

[90]  B. A. Simco,et al.  SEX DIFFERENTIATION OF CHANNEL CATFISH GONADS : NORMAL DEVELOPMENT AND EFFECTS OF TEMPERATURE , 1996 .

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

[92]  T. Pandian,et al.  Hormonal induction of sex reversal in fish , 1995 .

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

[94]  B. A. Simco,et al.  Feminization of channel catfish by oral administration of steroid sex hormones , 1983 .