Journal of Oceanology and Limnology

, Volume 36, Issue 6, pp 2337–2350 | Cite as

Pf-Dmrt4, a potential factor in sexual development in the pearl oyster Pinctada fucata

  • Qi Wang (王琦)
  • Yu Shi (石禹)
  • Maoxian He (何毛贤)Email author


The mechanisms of sex determination and sex differentiation in the pearl oyster Pinctada fucata are currently poorly understood. We therefore investigated the roles of orthologs of the Dmrt gene family, key players in male gonad differentiation in mammals, in P. fucata sex differentiation and sexual development. Pf-Dmrt4 exhibits features typical of the D mrt family, and displays significant homologies to the DMRT4 cluster. Pf-Dmrt4 mRNA expression in the gonads during a gametogenic cycle, measured by quantitative polymerase chain reaction, was maximal in mature individuals. Pf-Dmrt4 expression, demonstrated by in situ hybridization, was localized in the spermatozoa, spermatids, oocytes and vitellogenic oocytes. Knockdown of Pf-Dmrt4 with double-stranded RNA resulted in decreased mRNA expression levels. And Pf-Dmrt4-dsRNA-injected groups showed spawning-stage male gonads, with ruptured follicles and released spermatozoa. Our results enhance the understanding of sex determination and differentiation in P. fucata and suggest that Pf-Dmrt4 could be involved in male gonadal development, and maintenance of male gonadal function.


Pinctada fucata Pf-Dmrt4 gene expression sexual development 


Unable to display preview. Download preview PDF.

Unable to display preview. Download preview PDF.


  1. 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 ic and Genome Res earch, 101 (3–4): 295–301.CrossRefGoogle Scholar
  2. Burtis K C, Baker B S. 1989. Drosophila doublesex gene controls somatic sexual differentiation by producing alternatively spliced mRNAs encoding related sexspecific polypeptides. Cell, 56 (6): 997–1 010.CrossRefGoogle Scholar
  3. Cao J L, Chen J J, Wu T T, Gan X, Luo Y J. 2010. Molecular cloning and sexually dimorphic expression of DMRT4 gene in Oreochromis aureus. Molecular Biology Rep orts, 37 (6): 2 781–2 788.CrossRefGoogle Scholar
  4. Crémazy F, Berta P, Girard F. 2001. Genome–wide analysis of Sox genes in Drosophila melanogaster. Mech anisms of Dev elopment, 109 (2): 371–375.CrossRefGoogle Scholar
  5. Croll R P, Wang C D. 2007. Possible roles of sex steroids in the controlof reproduction in bivalve molluscs. Aquaculture, 272 (1–4): 76–86.CrossRefGoogle Scholar
  6. Du X D, Fan G Y, Jiao Y, Zhang H, Guo X M, Huang R L, Zheng Z, Bian C, Deng Y W, Wang Q H, Wang Z D, Liang X M, Liang H Y, Shi C C, Zhao X X, Sun F M, Hao R J, Bai J, Liu J L, Chen W B, Liang J L, Liu W Q, Xu Z, Shi Q, Xu X, Zhang G F, Liu X. 2017. The pearl oyster Pinctada fucata martensii genome and multi–omic analyses provide insights into biomineralization. Giga Science, 6 (8): 1–12.CrossRefGoogle Scholar
  7. Fabioux C, Pouvreau S, Le Roux F, Huvet A. 2004. The oyster vasa–like gene: a specific marker of the germline in Crassostrea gigas. Biochem ical and Biophys ical Res earch Commun ications, 315 (4): 897–904.CrossRefGoogle Scholar
  8. Feng Z F, Shao M Y, Sun D P, Zhang Z F. 2010. Cloning, characterization and expression analysis of Cf–dmrt4–like gene in Chlamys farreri. J ournal of Fish ery Sci ences of China, 17 (5): 930–940. (in Chinese with English abstract)Google Scholar
  9. Guo Y Q, Li Q, Gao S, Zhou X, He Y, Shang X, Cheng H H, Zhou R J. 2004. Molecular cloning, characterization, and expression in brain and gonad of Dmrt5 of zebrafish. Biochem ical and Biophys ical Res earch Commun ications, 324 (2): 569–575.CrossRefGoogle Scholar
  10. Hodgkin J. 2002. The remarkable ubiquity of DM domain factors as regulators of sexual phenotype: ancestry or aptitude? Genes & Dev elopment, 16 (18): 2 322–2 326.CrossRefGoogle Scholar
  11. Hong C S, Park B Y, Saint–Jeannet J P. 2007. The function of Dmrt genes in vertebrate development: it is not just about sex. Dev elopmental Biology, 310 (1): 1–9.CrossRefGoogle Scholar
  12. Klinbunga S, Amparyup P, Khamnamtong B, Hirono I, Aoki T, Jarayabhand P. 2009. Isolation and characterization of testis–specific DMRT1 in the tropical abalone ( Haliotis asinina ). Biochemical Genetics, 47 (1–2): 66–79.CrossRefGoogle Scholar
  13. Leveugle M, Prat K, Popovici C, Birnbaum D, Coulier F. 2004. Phylogenetic analysis of Ciona intestinalis gene superfamilies supports the hypothesis of successive gene expansions. J ournal of Molecular Evolution, 58 (2): 168–181.CrossRefGoogle Scholar
  14. Marı́n I, Baker B S. 1998. The evolutionary dynamics of sex determination. Science, 281 (5385): 1 990–1 994.CrossRefGoogle Scholar
  15. Miller S W, Hayward D C, Bunch T A, Miller D J, Ball E E, Bardwell V J, Zarkower D, Brower D L. 2003. A DM domain protein from a coral, Acropora millepora, homologous to proteins important for sex determination. Evolution & Dev elopment, 5 (3): 251–258.CrossRefGoogle Scholar
  16. Miyashita T, Hanashita T, Toriyama M, Takagi R, Akashika T, Higashikubo N. 2008. Gene cloning and biochemical characterization of the BMP–2 of Pinctada fucata. Bio s cience, Biotechnology, and Biochemistry, 72 (1): 37–47.CrossRefGoogle Scholar
  17. Naimi A, Martinez A S, Specq M L, 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 arative Biochem istry and Physio logy Part A: Molecular & Integr ative Physiology, 152 (2): 189–196.CrossRefGoogle Scholar
  18. 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 (3): 333–343.CrossRefGoogle Scholar
  19. Peng Q L, Pu Y G, Cheng Z H, Nie L W. 2005. Sequence analysis of three Dmrt genes in Macrobrachium rosenbergi i. J ournal of Fish ery Sci ences of China, 12 (1): 5–9. (in Chinese with English abstract)Google Scholar
  20. Raymond C S, Kettlewell J R, Hirsch B, Bardwell V J, Zarkower D. 1999. Expression of Dmrt1 in the genital ridge of mouse and chicken embryos suggests a role in vertebrate sexual development. Dev elopmental Biology, 215 (2): 208–220.CrossRefGoogle Scholar
  21. Raymond C S, Murphy M W, O’Sullivan M G, Bardwell V J, Zarkower D. 2000. Dmrt1, a gene related to worm and fly sexual regulators, is required for mammalian testis differentiation. Genes & Dev elopment, 14 (20): 2 587–2 595.CrossRefGoogle Scholar
  22. Shen X Y, Cui J Z, Yang G P, Gong Q L, Gu Q Q. 2007. Expression detection of DMRTs and two sox9 genes in Takifugu rubripes (Tetraodontidae, Vertebrata). J ournal of Ocean U niversity of China, 6 (2): 182–186.CrossRefGoogle Scholar
  23. Shen M M, Hodgkin J. 1988. M ab–3, a gene required for sexspecific yolk protein expression and a male–specific lineage in C. elegans. Cell, 54 (7): 1 019–1 031.CrossRefGoogle Scholar
  24. Shi Y, Wang Q, He M X. 2014. Molecular identification of dmrt2 and dmrt5 and effect of sex steroids on their expressions in Chlamys nobilis. Aqu a culture, 426–427 (1): 21–30.CrossRefGoogle Scholar
  25. Suzuki M, Saruwatari K, Kogure T, Yamamoto Y, Nishimura T, Kato T, Nagasawa H. 2009. An acidic matrix protein, Pif, is a key macromolecule for nacre formation. Science, 325 (5946): 1 388–1 390.CrossRefGoogle Scholar
  26. Takeuchi T, Kawashima T, Koyanagi R, Gyoja F, Tanaka M, Ikuta T, Shoguchi E, Fujiwara M, Shinzato C, Hisata K, Fujie M, Usami T, Nagai K, Maeyama K, Okamoto K, Aoki H, Ishikawa T, Masaoka T, Fujiwara A, Endo K, Endo H, Nagasawa H, Kinoshita S, Asakawa S, Watabe S, Satoh N. 2012. Draft genome of the pearl oyster Pinctada fucata: a platform for understanding bivalve biology. DNA Research, 19 (2): 117–130.CrossRefGoogle Scholar
  27. Teaniniuraitemoana V, Huvet A, Levy P, Klopp C, Lhuillier E, Gaertner–Mazouni N, Gueguen Y, Le Moullac G. 2014. Gonad transcriptome analysis of pearl oyster Pinctada margaritifera: identification of potential sex differentiation and sex determining genes. BMC Genomics, 15 (1): 491.CrossRefGoogle Scholar
  28. Volff J N, Zarkower D, Bardwell V J, Schartl M. 2003. Evolutionary dynamics of the DM domain gene family in metazoans. J ournal of Mo lecular Evolution, 57 (S1): S241–S249.CrossRefGoogle Scholar
  29. Wang C, Croll R P. 2003. Effects of sex steroids on in vitro gamete release in the sea scallop, Placopecten magellanicus. Invert ebrate Reprod uction & Dev elopment, 44 (2–3): 89–100.CrossRefGoogle Scholar
  30. Wen A Y, You F, Tan X G, Sun P, Ni J, Zhang Y Q, Xu D D, Wu Z H, Xu Y L, Zhang P J. 2009. Expression pattern of dmrt4 from olive flounder ( Paralichthys olivaceus ) in adult gonads and during embryogenesis. Fish Physiology and Biochem istry, 35 (3): 421–433.CrossRefGoogle Scholar
  31. Yamaguchi A, Lee K H, Fujimoto H, Kadomura K, Yasumoto S, Matsuyama M. 2006. Expression of the DMRT gene and its roles in early gonadal development of the Japanese pufferfish Takifugu rubripes. Comp arative Biochem istry and Physiology Part D: Genomics and Proteomics, 1 (1): 59–68.Google Scholar
  32. Ying M, Chen B, Tian Y H, Hou Y, Li Q, Shang X, Sun J H, Cheng H H, Zhou R J. 2007. Nuclear import of human sexual regulator DMRT1 is mediated by importin–β. Biochim ica e t Biophys ica Acta Molecular Cell Resaerch, 1773 (6): 804–813.CrossRefGoogle Scholar
  33. Yu F F, Gui J F, Zhou L, Wang M F, Yu X Y. 2009. Cloning and expression characterization of Dmrt5 in Pinctada martensii. Acta Hydrobiologica Sinica, 33 (5): 844–850. (in Chinese with English abstract)CrossRefGoogle Scholar
  34. Yu F F, Wang M F, Zhou L, Gui J F, Yu X Y. 2011. Molecular cloning and expression characterization of DMRT2 in Akoya pearl oysters, Pinctada martensii. J ournal of Shellfish Res earch, 30 (2): 247–254.CrossRefGoogle Scholar
  35. Yu F F, Yu X Y, Wang M F, Zhou L, Gui J F. 2007. Sex reversal phenomena in bivalves and its mechanism. Acta Hydrobiologica Sinica, 31 (4): 576–580. (in Chinese with English abstract)Google Scholar
  36. Zhang L X, Hua Z C, Ren J G, Meng A M. 2001. The nuclear localization signal of zebrafish terra is located within the DM domain. FEBS Letters, 503 (1): 25–29.CrossRefGoogle Scholar
  37. Zhang N, Xu F, Guo X M. 2014. Genomic analysis of the pacific oyster ( Crassostrea gigas ) reveals possible conservation of vertebrate sex determination in a mollusc. Genes, Genomes, Genetics, 4 (11): 2 207–2 217.Google Scholar

Copyright information

© Chinese Society for Oceanology and Limnology, Science Press and Springer-Verlag GmbH Germany, part of Springer Nature 2018

Authors and Affiliations

  • Qi Wang (王琦)
    • 1
    • 2
  • Yu Shi (石禹)
    • 1
  • Maoxian He (何毛贤)
    • 1
    Email author
  1. 1.CAS Key Laboratory of Tropical Marine Bio-resources and Ecology, Guangdong Provincial Key Laboratory of Applied Marine Biology, South China Sea Institute of OceanologyChinese Academy of SciencesGuangzhouChina
  2. 2.Taiyuan University of TechnologyTaiyuanChina

Personalised recommendations