Environmental Biology of Fishes

, Volume 75, Issue 2, pp 159–166 | Cite as

Plasma Sex Steroid Levels and Steroidogenesis in the Gonad of the Self-fertilizing Fish Rivulus marmoratus

  • Masako Minamimoto
  • Yoshitaka SakakuraEmail author
  • Kiyoshi Soyano
  • Yoshiaki Akaba
  • Atsushi Hagiwara


The mangrove killifish, Rivulus marmoratus, is the only known self-fertilizing vertebrate. This species is sexually dimorphic; sexually mature individuals are either hermaphrodite or primary and secondary males. Although the mangrove killifish has a unique reproductive strategy, there has been no study on the reproductive endocrinology of this species. Thus we investigated plasma sex steroid hormone levels and steroidogenesis in the gonads of R. marmoratus by enzyme linked immunosorbent assay (ELISA). Plasma 17β-estradiol (E2) and 11-ketotestosterone (11-KT) were detected both in hermaphrodite and in primary male. Ovarian follicles (follicle-enclosed oocytes) from hermaphrodites, which were categorized into early yolk stage and late yolk stage, and testis tissue of primary males were cultured with different concentrations of 17α-hydroxyprogesterone (OHP) or testosterone (T) for 24 h. Production of T, E2, 11-KT and 17α-20 β-dihydroxy-4-pregnen-3-one (17α,20β-P) in the medium from tissue culture were measured by ELISA. Early and late ovarian follicles of hermaphrodites and testis pieces of primary males synchronously secreted E2, 11-KT, and 17α,20β-P following incubation with OHP or T. We conclude that both hermaphrodite and primary male of the mangrove killifish secrete estrogen, androgen, and progestin synchronously.


mangrove killifish steroid hormone hermaphrodite tissue culture 


Unable to display preview. Download preview PDF.

Unable to display preview. Download preview PDF.


  1. Abel, D.C., Koenig, C.G., Davis, W.P. 1987Emersion in the mangrove forest fish Rivulus marmoratus: a unique response to hydrogen sulfideEnviron. Biol. Fish.186772CrossRefGoogle Scholar
  2. Asahina, K., Kanbegawa, A., Higashi, T. 1995Development of a microtiter plate enzyme-linked immunosorbent assay for 17α,20β-trihydroxy-4-pregnen-3-one, a teleost gonadal steroidFish. Sci.61491494Google Scholar
  3. Bradford, C.S., Taylor, M.H. 1987Semilunar changes in estradiol and cortisol coincident with gonadal maturation and spawning in the killifish Fundulus heteroclitus Gen. Comp. Endocrinol.667178CrossRefGoogle Scholar
  4. Cheek, A.O., Thomas, P., Sullivan, C.V. 2000Sex steroids relative to alternative mating behaviors in the simultaneous hermaphrodite Serranus subligarius (Perciformes: Serranidae)Horm. Behav.37198211CrossRefGoogle Scholar
  5. Cochran, R.C. 1987Serum androgens during the annual reproductive cycle of the male mummichog, Fundulus heteroclitus Gen. Comp. Endocrinol.65141148CrossRefGoogle Scholar
  6. Cole, K.S. & D.L.G. Noakes, 1997. Gonadal development and sexual allocation in mangrove killifish, Rivulus marmoratus (Pisces: Atherinomorpha). Copeia: 596 – 600Google Scholar
  7. Davis, W.P. 1988Reproductive and developmental responses in the self-fertilizing fish, Rivulus marmoratus, induced by the plasticizer, di-n-butylphthalateEnviron. Biol. Fish.218190CrossRefGoogle Scholar
  8. Dunson, W.A. & D.B. Dunson., 1999. Factors influencing growth and survival of the killifish, Rivulus marmoratus, held inside enclosures in mangrove swamps. Copeia: 661 – 668Google Scholar
  9. Frick, N.T., Wright, P.A. 2002Nitrogen metabolism and excretion in the mangrove killifish Rivulus marmoratus I The influence of environment salinity and external ammoniaThe J. Exp. Biol.2057989Google Scholar
  10. Grageda, M.V.C., Sakakura, Y., Hagiwara, A. 2004Early development of the self-fertilizing mangrove killifish, Rivulus marmoratus, reared in laboratoryIchthyol. Res.51309315CrossRefGoogle Scholar
  11. Grageda, M.V.C., Sakakura, Y., Minamimoto, M., Hagiwara, A. 2005Differences in life-history traits in two clonal strains of the self-fertilizing fish, Rivulus marmoratus Environ. Biol. Fish.73427436CrossRefGoogle Scholar
  12. Greeley, M.R., Calder, D.R.,Jr., Taylor, M.H., Hols, H., Wallace, R.A. 1986Oocyte maturation in the mummichog (Fundulus heteroclitus): Effects of steroids on germinal vesicle breakdown of intact follicles in vitro Gen. Comp. Endocrinol.62281289CrossRefGoogle Scholar
  13. Harrington, R.W.,Jr. 1961Oviparous hermaphroditic fish with internal self-fertilizationScience13417491750Google Scholar
  14. Harrington, R.W.,Jr 1967Environmentally controlled induction of primary male gonochorists from eggs of the self-fertilizing hermaphroditic fish, Rivulus marmoratus PoeyBiol. Bull132174199Google Scholar
  15. Harrington, R.W. Jr. 1971. How ecological and genetic factors interact to determine when self-fertilizing hermaphrodites of Rivulus marmoratus change into functional secondary males, with a reappraisal of the modes of intersexuality among fishes. Copeia 389 – 432Google Scholar
  16. Harrington, R.W.,Jr., Kallman, K.D. 1968The homozygosity of clones of the self-fertilizing hermaphroditic fish Rivulus marmoratus Poey (Cyprinodontidae, Atheriniformes)Am. Nat.102337343CrossRefGoogle Scholar
  17. Kallman, K.D., Harrington, R.W.,Jr. 1964Evidence for the existence of homozygous clones in the self-fertilizing hermaphroditic teleost Rivulus marmoratus (Poey)Biol. Bull.126101114Google Scholar
  18. Kobayashi, D., Tanaka, M, Fukuda, S., Nagahama, Y. 1996Steroidogenesis in the ovarian follicles of the Medaka (Oryzias latipes) during vitellogenesis and oocyte maturationZool. Sci.13921927CrossRefGoogle Scholar
  19. Kobayshi, M., Aida, K., Stacey, N.E. 1991Induction of testis development by implantation of 11-ketotestosterone in female goldfishZool. Sci.8389393Google Scholar
  20. Kobayashi, M., Furukawa, K., Kim, M.-H., Aida, K. 1997Induction of male-type gonadotropin secretion by implantation of 11-ketotetosterone in female goldfishGen. Comp. Endocrinol.108434445CrossRefGoogle Scholar
  21. Lin, H.-C., Dunson, W.A. 1993The effect of salinity on the acute toxicity of cadmium to the tropical, estuarine, hermaphroditic fish, Rivulus marmoratus: A comparison of Cd, Cu, and Zn tolerance with Fundulus heteroclitus Arch. Environ. Contamin. Toxicol.254147CrossRefGoogle Scholar
  22. Miura, T., Miura, C., Ohta, T., Nader, M.R., Todo, T., Yamauchi, K. 1999Estradiol-17β stimulates the renewal of spermatogonial stem cells in malesBiochem. Biophys. Res. Commun.264230234CrossRefGoogle Scholar
  23. Nagahama, Y. 1987Gonadotropin action on gametogenesis and steroidogenesis in teleost gonadsZool. Sci.4209222Google Scholar
  24. Nagahama, Y. 1994Endocrine regulation of gametogenesis in fishInt. J. Dev. Biol.38217229Google Scholar
  25. Nagahama, Y. 199717α,20β-Dihydroxy-4-pregnen-3-one, a maturation-inducing hormone in fish oocytes: Mechanisms of synthesis and actionSteroids62190196CrossRefGoogle Scholar
  26. Park, E.-H., Chang, H.-H., Joo, W.N. 1994Assessment of the estuarine hermaphroditic fish, Rivulus marmoratus as a useful euryhaline species for acute toxicity tests as shown using cadmiumCanadian J. Fish. Aqua. Sci.51280285CrossRefGoogle Scholar
  27. Petrino, T.R., Lin, Y.-W.P., Netherton, J.C., Powell, D.H., Wallace, R.A. 1993Steroidogenesis in Fundulus heteroclitus V: Purification, characterization, and metabolism of 17α,20β-dihydroxy-4-pregnen-3-one by intact follicles and its role in oocyte maturationGen. Comp. Endocrinol.92115CrossRefGoogle Scholar
  28. Sakai, N., Iwamatsu, T., Yamauchi, K., Suzuki, N., Nagahama, Y. 1988Influence of follicular development on steroid production in the medaka (Oryzias latipes) ovarian follicle in response to exogenous substratesGen. Comp. Endocrinol.71516523CrossRefGoogle Scholar
  29. Sakakura, Y., Noakes, D.L.G. 2000Age, growth, sexual development in the self-fertilizing hermaphroditic fish Rivulus marmoratus Environ. Biol. Fish.59309317CrossRefGoogle Scholar
  30. Soto, C.G., Leatherland, J.F., Noakes, D.L.G. 1992Gonadal histology in the self-fertilizing hermaphroditic fish Rivulus marmoratus (Pisces, Cyprinodontidae)Canadian J. Zool.7023382347CrossRefGoogle Scholar
  31. Soto, C.G., Noakes, D.L.G. 1994Colouration and gender in the hermaphroditic fish Rivulus marmoratus Poey (Teleostei: Rivulidae)Ichthyol. Explor. Freshw.57990Google Scholar
  32. Turner, B.J., Elder, J.F.,Jr., Laughlin, T.F., Davis, W.P. 1990Genetic variation in clonal vertebrates detected by simple-sequence DNA fingerprintingProc. Nat. Acad. Sci.8756535657Google Scholar

Copyright information

© Springer 2006

Authors and Affiliations

  • Masako Minamimoto
    • 1
  • Yoshitaka Sakakura
    • 2
    Email author
  • Kiyoshi Soyano
    • 3
  • Yoshiaki Akaba
    • 1
  • Atsushi Hagiwara
    • 1
  1. 1.Graduate School of Science and TechnologyNagasaki UniversityNagasakiJapan
  2. 2.Faculty of FisheriesNagasaki UniversityNagasakiJapan
  3. 3.Institute for East China Sea ResearchNagasaki UniversityNagasakiJapan

Personalised recommendations