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A novel Dmrt gene is specifically expressed in the testis of Chinese mitten crab, Eriocheir sinensis

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Dmrt is a family of genes related to the sexual regulators Doublesex of Drosophila melanogaster and Mab-3 of Caenorhabditis elegans. Dmrt genes are widely conserved and known for their involvement in sex determination and differentiation across phyla. In this study, we report here the identification of a novel Dmrt gene, named EsDmrt-like, from Chinese mitten crab Eriocheir sinensis. EsDmrt-like encodes a protein of 236 amino acids without intron. The protein contains a conserved DNA-binding DM domain that is characteristic of Dmrt genes. The DM domain shares 98% identity with that of Drosophila Dmrt99B and vertebrate Dmrt5, but outside the DM domain, there is little homology in sequence and no other conserved domain such as DMA specific to the Dmrt99B and Dmrt3–5. Interestingly, the expression pattern of EsDmrt-like is quite similar with that of vertebrate Dmrt1. Reverse transcription–polymerase chain reaction showed that EsDmrt-like transcripts were detectable only in testis with much higher expression at immature stage. In situ hybridization to gonad sections indicated that the EsDmrt-like mRNA was exclusively localized in Sertoli cells around the periphery of seminiferous tubules and developing germ cells including spermatogonium, spermatocyte, and spermatid, but absent in spermatozoa. This finding strongly suggests an essential role for EsDmrt-like in the male testicular development/differentiation of the crab.

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  1. Aflalo E, Hoang T, Nguyen V, Lam Q, Nguyen D, Trinh Q, Raviv S, Sagi A (2006) A novel two-step procedure for mass production of all-male populations of the giant freshwater prawn Macrobrachium rosenbergii. Aquaculture 256:468–478

  2. Aoyama S, Shibata K, Tokunaga S, Takase M, Matsui K, Nakamura M (2003) Expression of Dmrt1 protein in developing and in sex-reversed gonads of amphibians. Cytogenet Genome Res 101:295–301

  3. Baker BS, Ridge KA (1980) Sex and the single cell. I. On the action of major loci affecting sex determination in Drosophila melanogaster. Genetics 94:383–423

  4. Burtis KC, Baker BS (1989) Drosophila doublesex gene controls somatic sexual differentiation by producing alternatively spliced mRNAs encoding related sex-specific polypeptides. Cell 56:997–1010

  5. Charniaux-Cotton H (1954) Découverte chez un Crustacé Amphipode (Orchestia gammarella) d’une glande endocrine responsible de la différenciation des caractéres sexuels primaires et secondaires mâles. C R Acad Sci Paris 239:780–782

  6. De Grandi A, Calvari V, Bertini V, Bulfone A, Peverali G, Camerino G, Borsani G, Guioli S (2000) The expression pattern of a mouse doublesex-related gene is consistent with a role in gonadal differentiation. Mech Dev 90:323–326

  7. Erdman SE, Burtis KC (1993) The Drosophila doublesex proteins share a novel zinc finger related DNA binding domain. EMBO J 12:527–535

  8. Guan G, Kobayashi T, Nagahama Y (2000) Sexually dimorphic expression of two types of DM (Doublesex/Mab-3)-domain genes in a teleost fish, the Tilapia (Oreochromis niloticus). Biochem Biophys Res Commun 272:662–666

  9. Guo Y, Li Q, Gao S, Zhou X, He Y, Shang X, Cheng H, Zhou R (2004) Molecular cloning, characterization, and expression in brain and gonad of Dmrt5 of zebrafish. Biochem Biophys Res Commun 324:569–575

  10. Guo Y, Cheng H, Huang X, Gao S, Yu H, Zhou R (2005) Gene structure, multiple alternative splicing, and expression in gonads of zebrafish Dmrt1. Biochem Biophys Res Commun 330:950–957

  11. Hinsch GW (1980) Spermiogenesis in a hermit-crab. Coenobita clypeatus. II. Sertoli cells. Tissue Cell 12:255–262

  12. Huang X, Guo Y, Shui Y, Gao S, Yu H, Cheng H, Zhou R (2005) Multiple alternative splicing and differential expression of dmrt1 during gonad transformation of the rice field eel. Biol Reprod 73:1017–1024

  13. Kato Y, Kobayashi K, Oda S, Colbourn JK, Tatarazako N, Watanabe H, Iguchi T (2008) Molecular cloning and sexually dimorphic expression of DM-domain genes in Daphnia magna. Genomics 91:94–101

  14. Kettlewell JR, Raymond CS, Zarkower D (2000) Temperature-dependent expression of turtle Dmrt1 prior to sexual differentiation. Genesis 26:174–178

  15. Khalaila I, Katz T, Abdu U, Yehezkel G, Sagi A (2001) Effects of implantation of hypertrophied androgenic glands on sexual characters and physiology of the reproductive system in the female red claw crayfish, Cherax quadricarinatus. Gen Comp Endocrinol 121:242–249

  16. Kobayashi T, Matsuda M, Kajiura-Kobayashi H, Suzuki A, Saito N, Nakamoto M, Shibata N, Nagahama Y (2004) Two DM domain genes, DMY and DMRT1, involved in testicular differentiation and development in the medaka, Oryzias latipes. Dev Dyn 231:518–526

  17. Kumar S, Tamura K, Nei M (2004) MEGA3: integrated software for molecular evolutionary genetics analysis and sequence alignment. Brief Bioinform 5:150–163

  18. Lu H, Huang X, Zhang L, Guo Y, Cheng H, Zhou R (2007) Multiple alternative splicing of mouse Dmrt1 during gonadal differentiation. Biochem Biophys Res Commun 352:630–634

  19. MacDonald CC, Redondo JL (2002) Reexamining the polyadenylation signal: were we wrong about AAUAAA? Mol Cell Endocrinol 190:1–8

  20. Malecha SR, Nevin PA, Ha P, Barck LE, Lamadrid-Rose Y, Masuno S, Hedgecock D (1992) Sex-ratios and sex-determination in progeny from crosses of surgically sex-reversed freshwater prawns, Macrobrachium rosenbergii. Aquaculture 105:201–218

  21. Manor R, Aflalo ED, Segall C, Weil S, Azulay D, Ventura T, Sagi A (2004) Androgenic gland implantation promotes growth and inhibits vitellogenesis in Cherax quadricarinatus females held in individual compartments. Invertebr Reprod Dev 45:151–159

  22. Marchand O, Govoroun M, D’Cotta H, McMeel O, Lareyre J, Bernot A, Laudet V, Guiguen Y (2000) DMRT1 expression during gonadal differentiation and spermatogenesis in the rainbow trout, Oncorhynchus mykiss. Biochim Biophys Acta 1493:180–187

  23. Matsuda M, Nagahama Y, Shinomiya A, Sato T, Matsuda C, Kobayashi T, Morrey CE, Shibata N, Asakawa S, Shimizu N, Hori H, Hamaguchi S, Sakaizumi M (2002) DMY is a Y-specific DM-domain gene required for male development in the medaka fish. Nature 417:559–563

  24. Matsushita Y, Oshima Y, Nakamura M (2007) Expression of DMRT genes in the gonads of Rana rugosa during sex determination. Zoolog Sci 24:95–99

  25. Miller SW, Hayward DC, Bunch TA, Miller DJ, Ball EE, Bardwell VJ, Zarkower D, Brower DL (2003) A DM domain protein from a coral, Acropora millepora, homologous to proteins important for sex determination. Evol Dev 5:251–258

  26. Nagamine C, Knight AW, Maggenti A, Paxman G (1980a) Effects of androgenic gland ablation on male primary and secondary sexual characteristics in the Malaysian prawn, Macrobrachium rosenbergii (de Man) (Decapoda, Palaemonidae), with first evidence of induced feminization in a nonhermaphroditic decapod. Gen Comp Endocrinol 41:423–441

  27. Nagamine C, Knight AW, Maggenti A, Paxman G (1980b) Masculinization of female Macrobrachium rosenbergii (de Man) (Decapoda, Palaemonidae) by androgenic gland implantation. Gen Comp Endocrinol 41:442–457

  28. Naimi A, Martinez AS, Specq ML, Mrac A, Diss B, Mathieu M, Sourdaine P (2009) Identification and expression of a factor of the DM family in the oyster Crassostrea gigas. Comp Biochem Physiol A Mol Integr Physiol 152:189–196

  29. Ottolenghi C, Fellous M, Barbieri M, McElreavey K (2002) Novel paralogy relations among human chromosomes support a link between the phylogeny of doublesex-related genes and the evolution of sex determination. Genomics 79:333–343

  30. Qiu GF, Weber GM, Rexroad CE 3rd, Yao J (2008) Identification of RtGST-1, a novel germ cell-specific mRNA-like transcript predominantly expressed in early previtellogenic oocytes in rainbow trout (Oncorhynchus mykiss). Mol Reprod Dev 75:723–730

  31. Raymond CS, Shamu CE, Shen MM, Seifert KJ, Hirsch B, Hodgkin J, Zarkower D (1998) Evidence for evolutionary conservation of sex-determining genes. Nature 391:691–695

  32. Raymond CS, Kettlewell JR, Hirsch B, Bardwell VJ, Zarkower D (1999a) Expression of Dmrt1 in the genital ridge of mouse and chicken embryos suggests a role in vertebrate sexual development. Dev Biol 215:208–220

  33. Raymond CS, Parker ED, Kettlewell JR, Brown LG, Page DC, Kusz K, Jaruzelska J, Reinberg Y, Flejter WL, Bardwell VJ, Hirsch B, Zarkower D (1999b) A region of human chromosome 9p required for testis development contains two genes related to known sexual regulators. Hum Mol Genet 8:989–996

  34. Sagi A, Cohen D, Milner Y (1990) Effect of androgenic gland ablation on morphotypic differentiation and sexual characteristics of male freshwater prawns, Macrobrachium rosenbergii. Gen Comp Endocrinol 77:15–22

  35. Shen MM, Hodgkin J (1988) mab-3, a gene required for sex-specific yolk protein expression and a male-specific lineage in C. elegans. Cell 54:1019–1031

  36. Smith CA, McClive PJ, Western PS, Reed KJ, Sinclair AH (1999) Conservation of a sex-determining gene. Nature 402:601–602

  37. Smith CA, Katz M, Sinclair AH (2003) DMRT1 is upregulated in the gonads during female-to-male sex reversal in ZW chicken embryos. Biol Reprod 68:560–570

  38. Sreenivasulu K, Ganesh S, Raman R (2002) Evolutionarily conserved, DMRT1, encodes alternatively spliced transcripts and shows dimorphic expression during gonadal differentiation in the lizard. Calotes versicolor. Mech Dev 119(Suppl 1):S55–S64

  39. Strauss WM (1989) Preparation of genomic DNA from mammalian tissues. In: Ausubel FM, Brent R, Kingston RE, Moore DD, Seidman JG, Smith JA, Struhl K, Chanda VB (eds) Current protocol in molecular biology. Wiley, New York, pp 221–222

  40. Thompson JD, Higgins DG, Gibson TJ (1994) CLUSTAL W: improving the sensitivity of progressive multiple sequence alignment through sequence weighting, position-specific gap penalties and weight matrix choice. Nucleic Acids Res 22:4673–4680

  41. Yi W, Zarkower D (1999) Similarity of DNA binding and transcriptional regulation by Caenorhabditis elegans MAB-3 and Drosophila melanogaster DSX suggests conservation of sex determining mechanisms. Development 126:873–881

  42. Ying M, Chen B, Tian Y, Hou Y, Li Q, Shang X, Sun J, Cheng H, Zhou R (2007) Nuclear import of human sexual regulator DMRT1 is mediated by importin-beta. Biochim Biophys Acta 1773:804–813

  43. Zhao Y, Lu H, Yu H, Cheng H, Zhou R (2007) Multiple alternative splicing in gonads of chicken DMRT1. Dev Genes Evol 217:119–126

  44. Zhu L, Wilken J, Phillips NB, Narendra U, Chan G, Stratton SM, Kent SB, Weiss MA (2000) Sexual dimorphism in diverse metazoans is regulated by a novel class of intertwined zinc fingers. Genes Dev 14:1750–1764.

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This research was funded by the National Natural Science Foundation of China (project number 30972242), Shanghai Science and Technology Project (project number 09391911100), and the Marine Biology Program of Shanghai Leading Academic Discipline (project number J50701).

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Correspondence to Gao-Feng Qiu.

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Communicated by S. Roth

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Zhang, E., Qiu, G. A novel Dmrt gene is specifically expressed in the testis of Chinese mitten crab, Eriocheir sinensis . Dev Genes Evol 220, 151–159 (2010).

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  • Dmrt-like gene
  • Testis-specific expression
  • Testicular development
  • Decapod crustacean