Advertisement

Fisheries Science

, Volume 75, Issue 1, pp 137–144 | Cite as

Induction of spermatogenesis in Japanese eel by recombinant goldfish gonadotropins

  • Youichi Hayakawa
  • Hidekazu Nagaya
  • Hiroki Kaki
  • Komei Hotta
  • Makito Kobayashi
Original Article Biology

Abstract

We previously demonstrated the biological activities of single-chain recombinant gonadotropins (scGTHs) of goldfish Carrassius auratus follicle-stimulating hormone (scFSH) and luteinizing hormone (scLH), produced by a baculovirus–silkworm larvae system, by using in vivo bioassays with some fishes including Japanese eel Anguilla japonica. Among the bioassays, we succeeded in induction of spermatogenesis of sexually immature male Japanese eels by both scFSH and scLH, especially resulting in the occurrence of spermatozoa in scLH-administered males. However, those recombinant hormones did not induce enlargement of testes. In order to further confirm the potency of recombinant GTHs for use in aquaculture species, we administered scFSH and scLH to males of Japanese eel at higher dosage and frequency (eight times with 2–5 days interval) than those of the previous study (five or six times with 7 days intervals), including combination of scFSH and scLH administration (scFSH–scLH). Gonadosomatic indices (GSI) of scLH- and scFSH–scLH-administered males were larger than those of initial control males and of control males that were injected with saline. Enlargement of testes was also confirmed by measurement of testicular lobe size in scFSH-, scLH-, and scFSH–scLH-administered males. By histological observation, occurrence of spermatozoa was confirmed in scLH- and scFSH–scLH-administered eels. Although milt production was not induced, higher dosage and frequency of scGTH administration was effective in promoting testicular development of immature eels. Thus, single-chain fish GTHs produced by the baculovirus–silkworm larvae system could be a useful tool for promotion of gonadal maturation in aquaculture fishes.

Keywords

FSH Goldfish Gonadotropin Japanese eel LH Recombinant hormone Single-chain Spermatogenesis 

Notes

Acknowledgments

We thank Professor Robert W. Ridge, Department of Life Science, International Christian University, for critical reading of the manuscript. We also thank Dr. Goro Yoshizaki and Dr. Tetsuro Morita of the Department of Marine Sciences, Tokyo University of Marine Science and Technology, for their technical assistances and valuable advice. This study was supported in part by a Grants-in-Aid from the Japan Society for the Promotion of Science for Scientific Research (MK, YH) and grants from the Ministry of Education, Sports, Science, and Technology for the Academic Frontier Project (2003–2007), International Christian University.

References

  1. 1.
    Hayakawa Y, Morita T, Kitamura W, Kanda S, Banba A, Nagaya H, Hotta K, Sohn YC, Yoshizaki G, Kobayashi M (2008) Biological activities of single-chain goldfish follicle-stimulating hormone and luteinizing hormone. Aquaculture 274:408–415CrossRefGoogle Scholar
  2. 2.
    Parhar IS (2003) Gonadotropin-releasing hormone receptors: neuroendocrine regulators and neuromodulators. Fish Physiol Biochem 28:13–18CrossRefGoogle Scholar
  3. 3.
    Swanson P, Dickey JT, Campbell B (2003) Biochemistry and physiology of fish gonadotropins. Fish Physiol Biochem 28:53–59CrossRefGoogle Scholar
  4. 4.
    Patiño R (1997) Manipulations of the reproductive system of fishes by means of exogenous chemicals. Prog Fish Cult 59:118–128CrossRefGoogle Scholar
  5. 5.
    Zohar Y, Mylonas CC (2001) Endocrine manipulations of spawning in cultured fish: from hormones to genes. Aquaculture 197:99–136CrossRefGoogle Scholar
  6. 6.
    Kobayashi M, Morita T, Ikeguchi K, Yoshizaki G, Suzuki T, Watabe S (2006) In vivo biological activity of recombinant goldfish gonadotropins produced by baculovirus in silkworm larvae. Aquaculture 256:433–442CrossRefGoogle Scholar
  7. 7.
    Vischer HF, Granneman JCM, Linskens MHK, Schulz RW, Bogerd J (2003) Both recombinant African catfish LH and FSH are able to activate the African catfish FSH receptor. J Mol Endocrinol 31:133–140PubMedCrossRefGoogle Scholar
  8. 8.
    Kamei H, Ohira T, Yoshiura Y, Uchida N, Nagasawa H, Aida K (2003) Expression of a biologically active recombinant follicle stimulating hormone of Japanese eel, Anguilla japonica using methylotropic yeast, Pichia pastoris. Gen Comp Endocrinol 134:244–254PubMedCrossRefGoogle Scholar
  9. 9.
    Kasuto H, Levavi-Sivan B (2005) Production of biologically active tethered tilapia LHβα by the methylotropic yeast Pichia pastoris. Gen Comp Endocrinol 140:222–232PubMedCrossRefGoogle Scholar
  10. 10.
    Aizen J, Kasuto H, Golan M, Zakay H, Levavi-Sivan B (2007) Tilapia follicle-stimulating hormone (FSH): immunochemistry, stimulation by gonadotropin-releasing hormone, and effect of biologically active recombinant FSH on steroid secretion. Biol Reprod 76:692–700PubMedCrossRefGoogle Scholar
  11. 11.
    Zmora N, Kazeto Y, Kumar RS, Schulz RW, Trant JM (2007) Production of recombinant channel catfish (Ictalurus punctatus) FSH and LH in S2 Drosophila cell line and an indication of their different actions. J Endocrinol 194:407–416PubMedCrossRefGoogle Scholar
  12. 12.
    Meiri I, Zmora N, Elizur A (1999) Functional expression of recombinant seabream FSH and LH in baculovirus infected insect cells. In: Norberg B, Kjesbu OS, Taranger GL, Andersson E, Stefansson SO (eds) Proceedings of the sixth international symposium on the reproductive physiology of fish, 4–9 July. Institute of Marine Research and University of Bergen, p 488Google Scholar
  13. 13.
    Choi EJ, Ko H, Shim J, Kim MA, Sohn YC (2005) Expression of gonadotropin genes in Manchurian trout Brachymystax lenok and production of recombinant gonadotropins. Fish Sci 71:1193–1200CrossRefGoogle Scholar
  14. 14.
    Morita T, Yoshizaki G, Kobayashi M, Takeuchi T (2003) Production of biologically-active recombinant goldfish gonadotropins in transgenic rainbow trout. Fish Physiol Biochem 28:473–474CrossRefGoogle Scholar
  15. 15.
    Morita T, Yoshizaki G, Kobayashi M, Watabe S, Takeuchi T (2004) Fish eggs as bioreactors: the production of bioactive luteinizing hormone in transgenic trout embryos. Transgenic Res 13:551–557PubMedCrossRefGoogle Scholar
  16. 16.
    Ko H, Park WD, Kim DJ, Kobayashi M, Sohn YC (2007) Biological activities of recombinant Manchurian trout FSH and LH: their receptor specificity, steroidogenic and vitellogenic potencies. J Mol Endocrinol 28:99–111CrossRefGoogle Scholar
  17. 17.
    Narayan P, Wu C, Puett D (2000) Genetic engineering of single-chain gonadotropins and hormone-receptor fusion proteins. Methods 21:59–66PubMedCrossRefGoogle Scholar
  18. 18.
    Garcia-Campayo V, Boime I (2001) Novel recombinant gonadotropins. Trends Endocrinol Metab 12:72–77PubMedCrossRefGoogle Scholar
  19. 19.
    Yamada H, Satoh R, Yamashita T, Kambegawa A, Iwata M (1997) Development of a time-resolved fluoroimmunoassay (TR-FIA) for testosterone: measurement of serum testosterone concentrations after testosterone treatment in the rainbow trout (Oncorhynchus mykiss). Gen Comp Endocrinol 106:181–188PubMedCrossRefGoogle Scholar
  20. 20.
    Yamada H, Satoh R, Ogoh M, Takaji K, Fujimoto Y, Hakuba T, Chiba H, Kambegawa A, Iwata M (2002) Circadian Changes in serum concentrations of steroids in Japanese char Salvelinus leucomaenis at the stage of final maturation. Zool Sci 19:891–898PubMedCrossRefGoogle Scholar
  21. 21.
    Todd PR (1981) Morphometric changes, gonad histology, and fecundity estimates in migrating New Zealand freshwater eels (Anguilla sp.). N Z J Mar Freshwater Res 13:155–170CrossRefGoogle Scholar
  22. 22.
    Ohta H, Unuma T (2003) Induction of sperm maturation. In: Aida K, Tsukamoto K, Yamauchi K (eds) Eel biology. Springer, Tokyo, pp 319–329Google Scholar
  23. 23.
    Kamei H, Kaneko T, Aida K (2006) In vivo gonadotropic effects of recombinant Japanese eel follicle-stimulating hormone. Aquaculture 261:771–775CrossRefGoogle Scholar
  24. 24.
    Ohta T, Miyake H, Miura C, Kamei H, Aida K, Miura T (2007) Follicle-stimulating hormone induces spermatogenesis mediated by androgen production in Japanese eel, Anguilla japonica. Biol Reprod 77:970–977PubMedCrossRefGoogle Scholar
  25. 25.
    Kobayashi M, Sohn YC, Morita T, Yoshizaki G, Hayakawa Y, Nagaya H, Suzuki T (2008) Production of recombinant fish gonadotropins by baculovirus in silkworm larvae and their in vivo biological activities. In: Nakamura TK (ed) Aquaculture Research Progress. Nova Science Publishers, New York (in press)Google Scholar
  26. 26.
    Boime I, Ben-Menahem D (1999) Glycoprotein hormone structure-function and analog design. Rec Prog Horm Res 54:271–289PubMedGoogle Scholar
  27. 27.
    Misaki R, Nagaya H, Fujiyama K, Yanagihara I, Honda T, Seki T (2003) N-linked glycan structures of mouse interferon-beta produced by Bombyx mori larvae. Biochem Biophys Res Commun 311:979–986PubMedCrossRefGoogle Scholar

Copyright information

© The Japanese Society of Fisheries Science 2008

Authors and Affiliations

  • Youichi Hayakawa
    • 1
  • Hidekazu Nagaya
    • 2
  • Hiroki Kaki
    • 2
  • Komei Hotta
    • 3
  • Makito Kobayashi
    • 1
  1. 1.Department of Life ScienceInternational Christian UniversityMitakaJapan
  2. 2.Katakura Industries Co. LtdSayamaJapan
  3. 3.Marine Ecology Research InstituteKashiwazakiJapan

Personalised recommendations