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cDNA cloning and expression analysis of a vasa-like gene in Pacific bluefin tuna Thunnus orientalis

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Abstract

In this study, a Pacific bluefin tuna (Thunnus orientalis) homolog of the Drosophila vasa gene, BtVLG (bluefin tuna vasa-like gene), was cloned and characterized for use as a molecular marker for germ cells in this species. Analysis of the nucleotide sequence revealed that BtVLG comprises 2,394 bps with an open reading frame of 1,932 bps encoding 644 amino acids. The deduced amino acid sequence contained arginine-glycine or arginine-glycine-glycine motifs and eight conserved motifs belonging to the DEAD-box protein family. The BtVLG sequence showed high similarity to Drosophila vasa (69.1%), zebrafish vasa homolog (80.5%), and tilapia vasa homolog (91.2%). In adult tissues, the BtVLG transcripts were specifically detected in ovary and testis. In situ hybridization analysis showed that BtVLG messenger RNA (mRNA) was detected in oogonia and previtellogenic oocytes in the ovary. In the testis, while BtVLG mRNA was detected in spermatogonia, it was not detected in the primary and secondary spermatocytes, spermatids, spermatozoa or gonadal somatic cells. Consequently, consensus sequences, sequence similarity, and specific localization of BtVLG mRNA in the germ cells all suggest that BtVLG is the bluefin tuna vasa homolog of the Drosophila vasa gene. Further, BtVLG can be used as a molecular marker for bluefin tuna germ cells.

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References

  1. General Fisheries Commission for the Mediterranean International Commission for the Conservation of Atlantic Tunas (2005) Report of the third meeting of the Ad Hoc GFCM/ICCAT working group on sustainable bluefin tuna farming fattening practices in the Mediterranean. FAO Fisheries Report No 779. FAO, Rome

  2. Sawada Y, Okada T, Miyashita S, Murata O, Kumai H (2005) Completion of the Pacific bluefin tuna, Thunnus orientalis, life cycle under aquaculture conditions. Aquac Res 36:413–421

    Article  Google Scholar 

  3. Nakamura Y, Ando M, Seoka M, Kawasaki K, Tsukamasa Y (2006) The changes in proximate compositions and glycogen contents in the dorsal ordinary muscles of the full-cycle cultured Pacific bluefin tuna Thunnus orientalis occurring with growth. Fish Sci 72:1140–1146

    Article  CAS  Google Scholar 

  4. Okutsu T, Suzuki K, Takeuchi Y, Takeuchi T, Yoshizaki G (2006) Testicular germ cells can colonize sexually undifferentiated embryonic gonad and produce functional eggs in fish. Proc Natl Acad Sci USA 103:2725–2729

    Article  PubMed  CAS  Google Scholar 

  5. Takeuchi Y, Yoshizaki G, Takeuchi T (2003) Generation of live fry from intraperitoneally transplanted primordial germ cells in rainbow trout. Biol Reprod 69:1142–1149

    Article  PubMed  CAS  Google Scholar 

  6. Takeuchi Y, Yoshizaki G, Takeuchi T (2004) Biotechnology: surrogate broodstock produces salmonids. Nature 430:629–630

    Article  PubMed  CAS  Google Scholar 

  7. Okutsu T, Shikina S, Kanno M, Takeuchi Y, Yoshizaki G (2007) Production of trout offspring from triploid salmon parents. Science 317:1517

    Article  PubMed  CAS  Google Scholar 

  8. Raz E (2000) The function and regulation of vasa-like genes in germ-cell development. Genome Biol 1:reviews 1017.1–reviews 1017.6

    Article  Google Scholar 

  9. Yoon C, Kawakami K, Hopkins N (1997) Zebrafish vasa homologue RNA is localized to the cleavage planes of 2- and 4-cell-stage embryos and is expressed in the primordial germ cells. Development 124:3157–3165

    PubMed  CAS  Google Scholar 

  10. Olsen LC, Aasland R, Fjose A (1997) A vasa-like gene in zebrafish identifies putative primordial germ cells. Mech Dev 66:95–105

    Article  PubMed  CAS  Google Scholar 

  11. Yoshizaki G, Sakatani S, Tominaga H, Takeuchi T (2000) Cloning and characterization of a vasa-like gene in rainbow trout and its expression in the germ cell lineage. Mol Reprod Dev 55:364–371

    Article  PubMed  CAS  Google Scholar 

  12. Kobayashi T, Kajiura-Kobayashi H, Nagahama Y (2000) Differential expression of vasa homologue gene in the germ cells during oogenesis and spermatogenesis in a teleost fish, tilapia, Oreochromis niloticus. Mech Dev 99:139–142

    Article  PubMed  CAS  Google Scholar 

  13. Shinomiya A, Tanaka M, Kobayashi T, Nagahama Y, Hamaguchi S (2000) The vasa-like gene, olvas, identifies the migration path of primordial germ cells during embryonic body formation stage in the medaka, Oryzias latipes. Dev Growth Differ 42:317–326

    Article  PubMed  CAS  Google Scholar 

  14. Xu H, Gui J, Hong Y (2005) Differential expression of vasa RNA and protein during spermatogenesis and oogenesis in the gibel carp (Carassius auratus gibelio), a bisexually and gynogenetically reproducing vertebrate. Dev Dyn 233:872–882

    Article  PubMed  CAS  Google Scholar 

  15. Ye D, Lv D, Song P, Peng M, Chen Y, Guo M, Yang Q, Hu Y (2007) Cloning and characterization of a rice field eel vasa-like gene cDNA and its expression in gonads during natural sex transformation. Biochem Genet 45:211–224

    Article  PubMed  CAS  Google Scholar 

  16. Miyake A, Saito T, Kashiwagi N, Ando D, Yamamoto A, Suzuki T, Nakatsuji N, Nakatsuji T (2006) Cloning and pattern of expression of the shiro-uo vasa gene during embryogenesis and its roles in PGC development. Int J Dev Biol 50:619–625

    Article  PubMed  CAS  Google Scholar 

  17. Cardinali M, Gioacchini G, Candiani S, Pestarino M, Yoshizaki G, Carnevali O (2004) Hormonal regulation of vasa-like messenger RNA expression in the ovary of the marine teleost Sparus aurata. Biol Reprod 70:737–743

    Article  PubMed  CAS  Google Scholar 

  18. Thomopson JD, Higgins DG, Gibson TJ (1994) CLUSTALW. Nucleic Acids Res 22:4673–4680

    Article  Google Scholar 

  19. Saitou N, Nei M (1987) The neighbor-joining method: a new method for reconstructing phylogenetic trees. Mol Biol Evol 4:406–425

    PubMed  CAS  Google Scholar 

  20. Hay B, Jan LY, Jan YN (1988) A protein component of Drosophila polar granules is encoded by vasa and has extensive sequence similarity to ATP-dependent helicase. Cell 55:577–587

    Article  PubMed  CAS  Google Scholar 

  21. Lasko PF, Ashburner M (1988) The product of the Drosophila gene vasa is very similar to eukaryotic initiation factor-4A. Nature 335:611–617

    Article  PubMed  CAS  Google Scholar 

  22. Linder P, Lasko PF, Ashburner M, Leroy P, Nielsen PJ, Nishi K, Schnier J, Slonimski PP (1989) Birth of the D-E-A-D box. Nature 337:121–122

    Article  PubMed  CAS  Google Scholar 

  23. Pause A, Sonenberg N (1992) Mutational analysis of a DEAD box RNA helicase: the mammalian translation initiation factor eIF-4A. EMBO J 11:2643–2654

    PubMed  CAS  Google Scholar 

  24. Liang L, Diehl-Jones W, Lasko P (1994) Localization of vasa protein to the Drosophila pole plasm is independent of its RNA-binding and helicase activities. Development 120:1201–1211

    PubMed  CAS  Google Scholar 

  25. Kajiura-Kobayashi H, Kobayashi T, Nagahama Y (2005) Cloning of cDNAs and the differential expression of A-type cyclins and Dmc 1 during spermatogenesis in the Japanese eel, a teleost fish. Dev Dyn 232:1115–1123

    Article  PubMed  CAS  Google Scholar 

  26. Iwai T, Lee J, Yoshii A, Yokota T, Mita K, Yamashita M (2004) Changes in the expression and localization of cohesin subunits during meiosis in a non-mammalian vertebrate, the medaka fish. Gene Expr Patterns 4:495–504

    Article  PubMed  CAS  Google Scholar 

  27. Iwai T, Yoshii A, Yokota T, Sakai C, Hori H, Kanamori A, Yamashita M (2006) Structural components of the synaptonemal complex, SYCP1 and SYCP3, in the medaka fish Oryzias latipes. Exp Cell Res 312:2528–2537

    Article  PubMed  CAS  Google Scholar 

  28. Okutsu T, Yano A, Nagasawa K, Shikina S, Kobayashi T, Takeuchi Y, Yoshizaki G (2006) Manipulation of fish germ cell: visualization, cryopreservation and transplantation. J Reprod Dev 52:685–693

    Article  PubMed  Google Scholar 

  29. Yano A, Suzuki K, Yoshizaki G (2008) Flow-cytometric isolation of testicular germ cells from rainbow trout (Oncorhynchus mykiss) carrying the green fluorescent protein gene driven by trout vasa regulatory regions. Biol Reprod 78:151–158

    Article  PubMed  CAS  Google Scholar 

  30. Seoka M, Kato K, Toshihiko Kubo T, Mukai Y, Sakamoto W, Kumai H, Murata O (2007) Gonadal maturation of pacific bluefin tuna Thunnus orientalis in captivity. Aquac Sci 55:289–292

    Google Scholar 

  31. Sawada Y, Seoka M, Kato K, Tamura T, Nakatani M, Hayashi S, Okada T, Tose K, Miyashita S, Murata O, Kumai H (2007) Testes maturation of reared Pacific bluefin tuna Thunnus orientalis at two-plus years old. Fish Sci 73:1070–1077

    Article  CAS  Google Scholar 

  32. Corriero A, Medina A, Mylonas CC, Abascal FJ, Deflorio M, Aragón L, Bridges CR, Santamaria N, Heinisch G, Vassallo-Agius R, Belmonte A, Fauvel C, Garcia A, Gordin H, De Metrio G (2007) Histological study of the effects of treatment with gonadotropin-releasing hormone agonist (GnRHa) on the reproductive maturation of captive-reared Atlantic bluefin tuna (Thunnus thynnus L.). Aquaculture 272:675–686

    Article  CAS  Google Scholar 

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Authors and Affiliations

  1. Department of Marine Biosciences, Tokyo University of Marine Science and Technology, 4-5-7 Konan, Minato-ku, Tokyo, 108-8477, Japan

    Kazue Nagasawa, Misako Miwa, Kentaro Higuchi & Goro Yoshizaki

  2. Research Center for Advanced Fisheries Science and Technology, Tokyo University of Marine Science and Technology, 670 Banda, Tateyama, Chiba, 294-0308, Japan

    Yutaka Takeuchi

  3. Oita Marine Biological Technology Center, Nippon Suisan Kaisha, Ltd, 508-8 Ariakeura, Saiki, Oita, 876-1204, Japan

    Tetsuro Morita & Toru Mitsuboshi

  4. Nagasaki Prefectural Institute of Fisheries, 1551-4 Taira-cho, Nagasaki, 851-2213, Japan

    Kadoo Miyaki & Kazushi Kadomura

Authors
  1. Kazue Nagasawa
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  2. Yutaka Takeuchi
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  3. Misako Miwa
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  4. Kentaro Higuchi
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  7. Kadoo Miyaki
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  8. Kazushi Kadomura
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  9. Goro Yoshizaki
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Correspondence to Goro Yoshizaki.

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Nagasawa, K., Takeuchi, Y., Miwa, M. et al. cDNA cloning and expression analysis of a vasa-like gene in Pacific bluefin tuna Thunnus orientalis . Fish Sci 75, 71–79 (2009). https://doi.org/10.1007/s12562-008-0021-9

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  • DOI: https://doi.org/10.1007/s12562-008-0021-9

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