Anti-masculinization induced by aromatase inhibitors in adult female zebrafish

Springer Science and Business Media LLC - Tập 21 - Trang 1-10 - 2020
Lu Chen1,2, Li Wang1,2, Qiwei Cheng1,2, Yi-Xuan Tu1,2, Zhuang Yang1,2, Run-Ze Li1,2, Zhi-Hui Luo1,2, Zhen-Xia Chen1,2
1Hubei Key Laboratory of Agricultural Bioinformatics, College of Life Science and Technology, Huazhong Agricultural University, Wuhan, People’s Republic of China
2College of Biomedicine and Health, Huazhong Agricultural University, Wuhan, People’s Republic of China

Tóm tắt

Early sex differentiation genes of zebrafish remain an unsolved mystery due to the difficulty to distinguish the sex of juvenile zebrafish. However, aromatase inhibitors (AIs) could direct juvenile zebrafish sex differentiation to male and even induce ovary-to-testis reversal in adult zebrafish. In order to determine the transcriptomic changes of sex differentiation in juvenile zebrafish and early sex-reversal in adult zebrafish, we sequenced the transcriptomes of juvenile and adult zebrafish treated with AI exemestane (EM) for 32 days, when juvenile zebrafish sex differentiation finished. EM treatment in females up-regulated the expression of genes involved in estrogen metabolic process, female gamete generation and oogenesis, including gsdf, macf1a and paqr5a, while down-regulated the expression of vitellogenin (vtg) genes, including vtg6, vtg2, vtg4, and vtg7 due to the lower level of Estradiol (E2). Furthermore, EM-juveniles showed up-regulation in genes related to cell death and apoptosis, such as bcl2l16 and anax1c, while the control-juveniles exhibited up-regulation of genes involved in positive regulation of reproductive process and oocyte differentiation such as zar1 and zpcx. Moreover, EM-females showed higher enrichment than control females in genes involved in VEGF signaling pathway, glycosaminoglycan degradation, hedgehog signaling pathway, GnRH signaling pathway and steroid hormone biosynthesis. Our study shows anti-masculinization in EM-treated adult females but not in EM-treated juveniles. This may be responsible for the lower sex plasticity in adults than juveniles.

Tài liệu tham khảo

Wilson CA, High SK, McCluskey BM, Amores A, Yan YL, Titus TA, Anderson JL, Batzel P, Carvan MJ 3rd, Schartl M, et al. Wild sex in zebrafish: loss of the natural sex determinant in domesticated strains. Genetics. 2014;198(3):1291–308. Liew WC, Orban L. Zebrafish sex: a complicated affair. Brief Funct Genomics. 2014;13(2):172–87. Nagabhushana A, Mishra RK. Finding clues to the riddle of sex determination in zebrafish. J Biosci. 2016;41(1):145–55. Jorgensen A, Morthorst JE, Andersen O, Rasmussen LJ, Bjerregaard P. Expression profiles for six zebrafish genes during gonadal sex differentiation. Reprod Biol Endocrinol. 2008;6:25. Liu H, Todd EV, Lokman PM, Lamm MS, Godwin JR, Gemmell NJ. Sexual plasticity: a fishy tale. Mol Reprod Dev. 2017;84(2):171–94. Zhang Y, Zhang S, Lu H, Zhang L, Zhang W. Genes encoding aromatases in teleosts: evolution and expression regulation. Gen Comp Endocrinol. 2014;205:151–8. Kang H, Xiao X, Huang C, Yuan Y, Tang D, Dai X, Zeng X. Potent aromatase inhibitors and molecular mechanism of inhibitory action. Eur J Med Chem. 2018;143:426–37. Steele RE, Mellor LB, Sawyer WK, Wasvary JM, Browne LJ. In vitro and in vivo studies demonstrating potent and selective estrogen inhibition with the nonsteroidal aromatase inhibitor CGS 16949A. Steroids. 1987;50(1–3):147–61. Kitano T, Takamune K, Nagahama Y, Abe SI. Aromatase inhibitor and 17alpha-methyltestosterone cause sex-reversal from genetical females to phenotypic males and suppression of P450 aromatase gene expression in Japanese flounder (Paralichthys olivaceus). Mol Reprod Dev. 2000;56(1):1–5. Afonso LO, Wassermann GJ, Terezinha de Oliveira R. Sex reversal in Nile tilapia (Oreochromis niloticus) using a nonsteroidal aromatase inhibitor. J Exp Zool. 2001;290(2):177–81. Kitano T, Yoshinaga N, Shiraishi E, Koyanagi T, Abe S. Tamoxifen induces masculinization of genetic females and regulates P450 aromatase and Mullerian inhibiting substance mRNA expression in Japanese flounder (Paralichthys olivaceus). Mol Reprod Dev. 2007;74(9):1171–7. Dranow DB, Tucker RP, Draper BW. Germ cells are required to maintain a stable sexual phenotype in adult zebrafish. Dev Biol. 2013;376(1):43–50. Wong TT, Saito T, Crodian J, Collodi P. Zebrafish germline chimeras produced by transplantation of ovarian germ cells into sterile host larvae. Biol Reprod. 2011;84(6):1190–7. Takatsu K, Miyaoku K, Roy SR, Murono Y, Sago T, Itagaki H, Nakamura M, Tokumoto T. Induction of female-to-male sex change in adult zebrafish by aromatase inhibitor treatment. Sci Rep. 2013;3:3400. Giudici D, Ornati G, Briatico G, Buzzetti F, Lombardi P, di Salle E. 6-Methylenandrosta-1,4-diene-3,17-dione (FCE 24304): a new irreversible aromatase inhibitor. J Steroid Biochem. 1988;30(1–6):391–4. Uchida D, Yamashita M, Kitano T, Iguchi T. Oocyte apoptosis during the transition from ovary-like tissue to testes during sex differentiation of juvenile zebrafish. J Exp Biol. 2002;205(Pt 6):711–8. Paul-Prasanth B, Bhandari RK, Kobayashi T, Horiguchi R, Kobayashi Y, Nakamoto M, Shibata Y, Sakai F, Nakamura M, Nagahama Y. Estrogen oversees the maintenance of the female genetic program in terminally differentiated gonochorists. Sci Rep. 2013;3:2862. Cheng WY, Zhang Q, Schroeder A, Villeneuve DL, Ankley GT, Conolly R. Editor's highlight: computational modeling of plasma Vitellogenin alterations in response to aromatase inhibition in fathead minnows. Toxicol Sci. 2016;154(1):78–89. Byrne BM, Gruber M, Ab G. The evolution of egg yolk proteins. Prog Biophys Mol Biol. 1989;53(1):33–69. Lin Q, Mei J, Li Z, Zhang X, Zhou L, Gui JF. Distinct and cooperative roles of amh and dmrt1 in self-renewal and differentiation of male germ cells in Zebrafish. Genetics. 2017;207(3):1007–22. Sambroni E, Lareyre JJ, Le Gac F. Fsh controls gene expression in fish both independently of and through steroid mediation. PLoS One. 2013;8(10):e76684. Huff M, da Silveira WA, Carnevali O, Renaud L, Hardiman G. Systems analysis of the liver Transcriptome in adult male Zebrafish exposed to the plasticizer (2-Ethylhexyl) phthalate (DEHP). Sci Rep. 2018;8(1):2118. Harada N, Honda SI, Hatano O. Aromatase inhibitors and enzyme stability. Endocr Relat Cancer. 1999;6(2):211–8. Kortner TM, Rocha E, Arukwe A. Previtellogenic oocyte growth and transcriptional changes of steroidogenic enzyme genes in immature female Atlantic cod (Gadus morhua L.) after exposure to the androgens 11-ketotestosterone and testosterone. Comp Biochem Physiol A Mol Integr Physiol. 2009;152(3):304–13. Krovel AV, Olsen LC. Sexual dimorphic expression pattern of a splice variant of zebrafish vasa during gonadal development. Dev Biol. 2004;271(1):190–7. Shen S, Park JW, Lu ZX, Lin L, Henry MD, Wu YN, Zhou Q, Xing Y. rMATS: robust and flexible detection of differential alternative splicing from replicate RNA-Seq data. Proc Natl Acad Sci U S A. 2014;111(51):E5593–601. Uchida D, Yamashita M, Kitano T, Iguchi T. An aromatase inhibitor or high water temperature induce oocyte apoptosis and depletion of P450 aromatase activity in the gonads of genetic female zebrafish during sex-reversal. Comp Biochem Physiol A Mol Integr Physiol. 2004;137(1):11–20. Fenske M, Segner H. Aromatase modulation alters gonadal differentiation in developing zebrafish (Danio rerio). Aquat Toxicol. 2004;67(2):105–26. Shi H, Gao T, Liu Z, Sun L, Jiang X, Chen L, Wang D. Blockage of androgen and administration of estrogen induce transdifferentiation of testis into ovary. J Endocrinol. 2017;233(1):65–80. Li M, Sun L, Wang D. Roles of estrogens in fish sexual plasticity and sex differentiation. Gen Comp Endocrinol. 2019;277:9–16. Sun D, Zhang Y, Wang C, Hua X, Zhang XA, Yan J. Sox9-related signaling controls zebrafish juvenile ovary-testis transformation. Cell Death Dis. 2013;4:e930. Muth-Kohne E, Westphal-Settele K, Bruckner J, Konradi S, Schiller V, Schafers C, Teigeler M, Fenske M. Linking the response of endocrine regulated genes to adverse effects on sex differentiation improves comprehension of aromatase inhibition in a fish sexual development test. Aquat Toxicol. 2016;176:116–27. Zhong L, Yuan L, Rao Y, Li Z, Zhang X, Liao T, Xu Y, Dai H. Distribution of vitellogenin in zebrafish (Danio rerio) tissues for biomarker analysis. Aquat Toxicol. 2014;149:1–7. Gavrieli Y, Sherman Y, Ben-Sasson SA. Identification of programmed cell death in situ via specific labeling of nuclear DNA fragmentation. J Cell Biol. 1992;119(3):493–501. Ewels P, Magnusson M, Lundin S, Kaller M. MultiQC: summarize analysis results for multiple tools and samples in a single report. Bioinformatics. 2016;32(19):3047–8. Kim D, Langmead B, Salzberg SL. HISAT: a fast spliced aligner with low memory requirements. Nat Methods. 2015;12(4):357–60. Li H, Handsaker B, Wysoker A, Fennell T, Ruan J, Homer N, Marth G, Abecasis G, Durbin R. Genome project data processing S: the sequence alignment/map format and SAMtools. Bioinformatics. 2009;25(16):2078–9. Anders S, Pyl PT, Huber W. HTSeq--a Python framework to work with high-throughput sequencing data. Bioinformatics. 2015;31(2):166–9. Love MI, Huber W, Anders S. Moderated estimation of fold change and dispersion for RNA-seq data with DESeq2. Genome Biol. 2014;15(12):550. Chen ZX, Oliver B. X Chromosome and Autosome Dosage Responses in Drosophila melanogaster Heads. G3 (Bethesda). 2015;5(6):1057–63. Ingleby FC, Flis I, Morrow EH. Sex-biased gene expression and sexual conflict throughout development. Cold Spring Harb Perspect Biol. 2014;7(1):a017632. Gentleman RC, Carey VJ, Bates DM, Bolstad B, Dettling M, Dudoit S, Ellis B, Gautier L, Ge Y, Gentry J, et al. Bioconductor: open software development for computational biology and bioinformatics. Genome Biol. 2004;5(10):R80. Yu G, Wang LG, Han Y, He QY. clusterProfiler: an R package for comparing biological themes among gene clusters. OMICS. 2012;16(5):284–7. Subramanian A, Tamayo P, Mootha VK, Mukherjee S, Ebert BL, Gillette MA, Paulovich A, Pomeroy SL, Golub TR, Lander ES, et al. Gene set enrichment analysis: a knowledge-based approach for interpreting genome-wide expression profiles. Proc Natl Acad Sci U S A. 2005;102(43):15545–50. Mootha VK, Lindgren CM, Eriksson KF, Subramanian A, Sihag S, Lehar J, Puigserver P, Carlsson E, Ridderstrale M, Laurila E, et al. PGC-1alpha-responsive genes involved in oxidative phosphorylation are coordinately downregulated in human diabetes. Nat Genet. 2003;34(3):267–73. rmats2sashimiplot [https://github.com/Xinglab/rmats2sashimiplot]. Accessed 1 May 2019.