Skip to main content
Log in

Cloning, characterization and function analysis of DAX1 in Chinese loach (Paramisgurnus dabryanus)

  • Original Paper
  • Published:
Genetica Aims and scope Submit manuscript

Abstract

The mechanisms of sex determination and differentiation have not been elucidated in most fish species. In this study, the full-length cDNAs of DAX1 was cloned and characterized in aquaculture fish Chinese loach (Paramisgurnus dabryanus), designated as Pd-DAX1. The cDNA sequence of Pd-DAX1 was 1261 bp, including 795 bp open reading frame (ORF) encoding 264 amino acids. Pd-DAX1 shares highly identical sequence with DAX1 homologues from different species. The expression profiles of Pd-DAX1 in different developmental stages and diverse adult tissues were analyzed by quantitative real-time RT-PCR and in situ hybridization (ISH). Pd-DAX1 was continuously expressed during embryogenesis, with the extensive distribution in the development of the central nervous system. Tissue distribution analysis revealed that Pd-DAX1 expressed widely in adult tissues, with the highest expression level found in testis, moderate level in ovary, showing a sex-dimorphic expression pattern. Pd-DAX1 mainly located in spermatogonia cells, spermatocytes, primary oocytes and previtellogenic oocyte cells, implying that Pd-DAX1 may involve in gametogenesis. These preliminary findings suggest that Pd-DAX1 gene is highly conserved during vertebrate evolution and involved in a wide range of developmental processes including embryogenesis, central nervous system development and gonad development.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Institutional subscriptions

Fig. 1
Fig. 2
Fig. 3
Fig. 4
Fig. 5
Fig. 6
Fig. 7

Similar content being viewed by others

References

  • Agoulnik IU, Krause WC, Bingman WE, Rahman HT, Amrikachi M, Ayala GE et al (2003) Repressors of androgen and progesterone receptor action. J Biol Chem 278:31136–31148

    Article  CAS  Google Scholar 

  • Anonymous (2012) Guidelines for the treatment of animals in behavioural research and teaching. Anim Behav 83:301–309

    Article  Google Scholar 

  • Arnold K, Bordoli L, Kopp J, Schwede T (2006) The SWISS-MODEL Workspace: a web-based environment for protein structure homology modelling. Bioinformatics 22:195–201

    Article  CAS  Google Scholar 

  • Bae D, Schaefer M, Partan B, Muglia L (1996) Characterization of the mouse DAX-1 gene reveals evolutionary conservation of a unique amino-terminal motif and widespread expression in mouse tissue. Endocrinology 137:3921–3927

    Article  CAS  Google Scholar 

  • Bertrand S, Thisse B, Tavares R, Sachs L, Chaumot A, Bardet PL, Escrivà H, Duffraisse M, Marchand O, Safi R, Thisse C, Laudet V (2007) Unexpected novel relational links uncovered by extensive developmental profiling of nuclear receptor expression. Plos Genet 3:e188

    Article  Google Scholar 

  • Chen S, Zhang H, Wang F, Zhang W, Peng G (2016) nr0b1 (DAX1) mutation in zebrafish causes female-to-male sex reversal through abnormal gonadal proliferation and differentiation. Mol Cell Endocrinol 433:105–116

    Article  CAS  Google Scholar 

  • Feng ZF, Zhang ZF, Shao MY, Zhu W (2011) Developmental expression pattern of the Fc-vasa-like gene, gonadogenesis and development of germ cell in Chinese shrimp, Fenneropenaeus chinensis. Aquaculture 314:202–209

    Article  CAS  Google Scholar 

  • Gao J, Wang J, Jiang J, Fan L, Wang W, Liu J, Zhang Q, Wang X (2013) Identification and characterization of a nanog homolog in Japanese flounder (Paralichthys olivaceus). Gene 531:411–421

    Article  CAS  Google Scholar 

  • Gasteiger E, Hoogland C, Gattiker A, Duvaud S, Wilkins MR, Appel RD, Bairoch A (2005) Protein identification and analysis tools on the ExPASy server. In: John MW (ed) The proteomics protocols handbook. Humana Press, New York, pp 571–607

    Chapter  Google Scholar 

  • Geourjon C, Deleage G (1995) SOPMA: significant improvements in protein secondary structure prediction by consensus prediction from multiple alignments. Comput Appl Biosci 11:681–684

    CAS  PubMed  Google Scholar 

  • Gurates B, Sebastian S, Yang S, Zhou J, Tamura M, Fang Z et al (2002) WT1 and DAX-1 inhibit aromatase P450 expression in human endometrial and endometriotic stromal cells. J Clin Endocrinol Metab 87:4369–4377

    Article  CAS  Google Scholar 

  • Haugen T, Almeida FF, Andersson E et al (2012) Sex differentiation in Atlantic cod (Gadus morhua L.): morphological and gene expression studies. Reprod Biol Endocrinol 10:47

    Article  CAS  Google Scholar 

  • Holter E, Kotaja N, Makela S, Strauss L, Kietz S, Janne OA, Gustafsson JA, Palvimo JJ, Treuter E (2002) Inhibition of androgen receptor (AR) function by the reproductive orphan nuclear receptor DAX-1. Mol Endocrinol 16:515–528

    Article  CAS  Google Scholar 

  • Hu Q, Guo W, Gao Y, Tang R, Li D (2015) Molecular cloning and characterization of amh and dax1 genes and their expression during sex inversion in rice-field eel Monopterus albus. Sci Rep 5:16667

    Article  CAS  Google Scholar 

  • Ito M, Yu R, Jameson JL (1997) DAX-1 inhibits SF-1-mediated transactivation via a carboxy-terminal domain that is deleted in adrenal hypoplasia congenita. Mol Cell Biol 17:1476–1483

    Article  CAS  Google Scholar 

  • Iyer AK, McCabe ER (2004) Molecular mechanisms of DAX1 action. Mol Genet Metab 83:60–73

    Article  CAS  Google Scholar 

  • Kawajiri K, Ikuta T, Suzuki T, Kusaka M, Muramatsu M, Fujieda K et al (2003) Role of the LXXLL-motif and activation function 2 domain in subcellular localization of Dax-1 (dosage-sensitive sex reversal-adrenal hypoplasia congenita critical region on the X chromosome, gene 1). Mol Endocrinol 17:994–1004

    Article  CAS  Google Scholar 

  • Kubista M, Andrade JM, Bengtsson M, Forootan A, Jonák J, Lind K et al (2006) The real-time polymerase chain reaction. Mol Aspects Med 27:95–125

    Article  CAS  Google Scholar 

  • Lalli E, Sassone-Corsi P (2003) DAX-1, an unusual orphan receptor at the crossroads of steroidogenic function and sexual differentiation. Mol Endocrinol 17:1445–1453

    Article  CAS  Google Scholar 

  • Lalli E, Bardoni B, Zazopoulos E, Wurtz JM, Strom TM, Moras D, Sas- sone-Corsi P (1997) A transcriptional silencing domain in DAX-1 whose mutation causes adrenal hypoplasia congenita. Mol Endocrinol 11(13):1950–1960

    Article  CAS  Google Scholar 

  • Li M, Wang L, Wang H, Liang H, Zheng Y, Qin F et al (2013) Molecular cloning and characterization of amh, dax1 and cyp19a1a genes and their response to 17α -methyltestosterone in pengze crucian carp. Comp Biochem Physiol Part C 157:372–381

    CAS  Google Scholar 

  • Li H, Liu J, Huang X, Wang D, Zhang Z (2014) Characterization, expression and function analysis of DAX1 gene of scallop (Chlamys farreri Jones and Preston 1904) during its gametogenesis. Ocean Coast Sea Res 13(4):696–704

    CAS  Google Scholar 

  • Livak KJ, Schmittgen TD (2001) Analysis of relative gene expression data using real time quantitative PCR and the 2-∆∆Ct method. Methods 25:402–408

    Article  CAS  Google Scholar 

  • Martins RS, Deloffre LA, Mylonas CC, Power DM, Canário AV (2007) Developmental expression of DAX1 in the European sea bass, Dicentrarchus labrax: lack of evidence for sexual dimorphism during sex differentiation. Reprod Biol Endocrinol 5(19):1–13

    Google Scholar 

  • McCabe E (2001) Adrenal hypoplasias and aplasias. The metabolic and molecular bases of inherited disease. McGraw-Hill, New York, pp 4263–4274

    Google Scholar 

  • McCabe ER (2007) DAX1: increasing complexity in the roles of this novel nuclear receptor. Mol Cell Endocrinol 265:179–182

    Article  Google Scholar 

  • Meeks JJ, Crawford SE, Russell TA, Morohashi K, Weiss J, Jameson JL (2003) Dax1 regulates testis cord organization during gonadal differentiation. Development 130:1029–1036

    Article  CAS  Google Scholar 

  • Nagl F, Schönhofer K, Seidler B, Mages J, Allescher HD, Schmid RM et al (2009) Retinoic acid-induced nNOS expression depends on a novel PI3K/Akt/DAX1 pathway in human TGW-nu-I neuroblastoma cells. Am J Physiol Cell Physiol 297:1146–1156

    Article  Google Scholar 

  • Nakamoto M, Wang DS, Suzuki A, Matsuda M, Nagahama Y, Shibata N (2007) Dax1 suppresses P450arom expression in medaka ovarian follicles. Mol Reprod Dev 74:1239–1246

    Article  CAS  Google Scholar 

  • Niakan KK, Davis EC, Clipsham RC, Jiang M, Dehart DB, Sulik KK et al (2006) Novel role for the orphan nuclear receptor Dax1 in embryogenesis, different from steroidogenesis. Mol Genet Metab 88:261–271

    Article  CAS  Google Scholar 

  • Nielsen H (2017) Predicting secretory proteins with signalp. Methods Mol Biol 1611:59–73

    Article  CAS  Google Scholar 

  • Parma P, Pailhoux E, Puissant C, Cotinot C (1997) Porcine Dax-1 gene: isolation and expression during gonadal development. Mol Cell Endocrinol 135:49–58

    Article  CAS  Google Scholar 

  • Peng N, Kim JW, Rainey WE, Carr BR, Attia GR (2003) The role of the orphan nuclear receptor, liver receptor homologue-1, in the regulation of human corpus luteum 3β -hydroxysteroid dehydrogenase type II. J Clin Endocrinol Metab 88:6020–6028

    Article  CAS  Google Scholar 

  • Pfall MW (2001) A new mathematical model for relative quantification in real-time RT-PCR. Nucleic Acids Res 29:2002–2007

    Google Scholar 

  • Powers J, Zhao Y, Lin S, Edward RB, McCabe (2009) The expression of nr0b1, the earliest gene in zebrafish tooth development, is a marker for human tooth and ameloblastoma formation. Dev Genes Evol 219:419–425

    Article  CAS  Google Scholar 

  • Sievers F, Wilm A, Dineen D, Gibson TJ, Karplus K, Li W, Lopez R, McWilliam H, Remmert M, Soding J, Thompson JD, Higgins DG (2011) Fast, scalable generation of high-quality protein multiple sequence alignments using Clustal Omega. Mol Syst Biol 7:539

    Article  Google Scholar 

  • Sim EU, Smith A, Szilagi E, Rae F, Ioannou P, Lindsay MH et al (2002) Wnt-4 regulation by the Wilms’ tumour suppressor gene, WT1. Oncogene 21:2948–2960

    Article  CAS  Google Scholar 

  • Small BC, Murdock CA, Bilodeau-Bourgeois AL, Peterson BC, Waldbieser GC (2008) Stability of reference genes for real-time PCR analyses in channel catfish (Ictalurus punctatus) tissues under varying physiological conditions. Comp Biochem Physiol B Biochem Mol Biol 151:296–304

    Article  Google Scholar 

  • Smith C, Clifford V, Western PS, Wilcox SA, Bell KS, Sinclair AH (2000) Cloning and expression of a DAX1 homologue in the chicken embryo. J Mol Endocrinol 24:23–32

    Article  CAS  Google Scholar 

  • Sugita J, Takase M, Nakamura M (2001) Expression of Dax-1 during gonadal development of the frog. Gene 280:67–74

    Article  CAS  Google Scholar 

  • Suzuki T, Kasahara M, Yoshioka H, Morohashi KI, Umesono K (2003) LXXLL-related motifs in Dax-1 have target specificity for the orphan nuclear receptors Ad4BP/SF-1 and LRH-1. Mol Cell Biol 23:238–249

    Article  CAS  Google Scholar 

  • Swain A, Zanaria E, Hacker A, Lovell-Badge R, Camerino G (1996) Mouse Dax1 expression is consistent with a role in sex determination as well as in adrenal and hypothalamus function. Nat Genet 12:404–409

    Article  CAS  Google Scholar 

  • Swain A, Narvaez V, Burgoyne P, Camerino G, Lovell-Badge R (1998) Dax1 antagonizes Sry action in mammalian sex determination. Nature 391:761–767

    Article  CAS  Google Scholar 

  • Tamura K, Stecher G, Peterson D, Filipski A, Kumar S (2013) MEGA6: molecular evolutionary genetics analysis version 6.0. Mol Biol Evol 30:2725–2729

    Article  CAS  Google Scholar 

  • Von Schalburg KR, Yasuike M, Yazawa R, de Boer JG, Reid L, So S et al (2011) Regulation and expression of sexual differentiation factors in embryonic and extragonadal tissues of Atlantic salmon. BMC Genom 12:31

    Article  Google Scholar 

  • Wang DS, Kobayashi T, Senthilkumaran B et al (2002) Molecular cloning of DAX1 and SHP c DNAs and their expression patterns in the Nile tilapia, Oreochromis niloticus. Biochem Biophys Res Commun 297(3):632–640

    Article  CAS  Google Scholar 

  • Yajima Y, Vestergaard MDC, Takagi M (2012) Damage in brain development by morpholino knockdown of zebrafish dax1. J Biosci Bioeng 113:683–688

    Article  CAS  Google Scholar 

  • Zanaria E, Muscatelli F, Bardoni B, Strom TM, Guioli S, Guo W, Lalli E, Moser C, Walker AP, McCabe ERB, Meitinger T, Monaco AP, Sassone-Corsi P, Camerino G (1994) An unusual member of the nuclear hormone receptor superfamily responsible for X-linked adrenal hypoplasia congenita. Nature 372:635–641

    Article  CAS  Google Scholar 

  • Zhang H, Thomsen JS, Johansson L, Gustafsson JA, Treuter E (2000) DAX-1 functions as an LXXLL-containing corepressor for activated estrogen receptors. J Biol Chem 275:39855–39859

    Article  CAS  Google Scholar 

  • Zhao Y, Yang Z, Phelan JK, Wheeler DA, Lin S, McCabe ERB (2006) Zebrafish dax1 is required for development of the interrenal organ, the adrenal cortex equivalent. Mol Endocrinol 20(11):2630–2640

    Article  CAS  Google Scholar 

Download references

Funding

This work is supported by grants from the National Natural Science Foundation of China (no. 31200923), Young Core Instructor Foundation of Henan Normal University (no. 5101049470610) and Program for Innovative Research Team (in Science and Technology) in University of Henan Province (no. 17IRTSTHN017).

Author information

Authors and Affiliations

Authors

Corresponding author

Correspondence to Xiaohua Xia.

Ethics declarations

Conflict of interest

The author(s) declare that they have no competing interests.

Human and animal rights statement

Animal maintenance and handling procedures followed the recommendations of the Association of Animal Behaviour and national regulations (Anonymous 2012).

Electronic supplementary material

Rights and permissions

Reprints and permissions

About this article

Check for updates. Verify currency and authenticity via CrossMark

Cite this article

Xia, X., Huo, W., Wan, R. et al. Cloning, characterization and function analysis of DAX1 in Chinese loach (Paramisgurnus dabryanus). Genetica 146, 487–496 (2018). https://doi.org/10.1007/s10709-018-0039-6

Download citation

  • Received:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s10709-018-0039-6

Keywords

Navigation