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Hydrobiologia

, Volume 365, Issue 1–3, pp 281–285 | Cite as

Ecdysteroids in nemerteans: further characterization and identification

  • Robert K. Okazaki
  • Mark J. Snyder
  • Casey C. Grimm
  • Ernest S. Chang
Article

Abstract

Ecdysteroids are a class of steroidal hormones thatare important in molting and reproduction inarthropods. These hormones have been recentlydetected in non-arthropodan groups, such assoft-bodied worms. To continue our efforts todetermine the presence of ecdysteroids in nemerteans,this study further documents the identification of20-hydroxyecdysone (20E), in Paranemertesperegrina, by gas chromatography/mass spectrometry(GC/MS). After C18 Sep-Pak extraction,radioimmunoassay of the 40% and 80% methanolicfractions from an extract of 1000 worms indicated 98and 156 ng of ecdysteroids, respectively. Ecdysteroids of these two samples, as well as the 20Eand ecdysone standards, were N-trimethylsilyimidazole(TMSI)-derivatized before GC/MS analysis. Themethanolic samples contained a large number ofcompounds but only small, insignificant peaks in thearea where ecdysteroid standards eluted. However, thereconstruction ion current (RIC) chromatograms for m/z561 indicated the presence of 20E at the correctretention time of 12.48 min in the 40% methanolicfraction. Reanalysis of the samples under selectedion monitoring mode demonstrated the presence of 20Ein both the 40% and 80% fractions. These resultsdemonstrate conclusively the presence of an activeecdysteroid in the phylum Nemertea.

nemertean Paranemertesperegrina ecdysteroid GC-MS RIA 

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References

  1. Barker, G. C., D. J. Chitwood & H. H. Rees, 1990. Ecdysteroids in helminths and annelids. Invert. Reprod. Dev. 18: 1–11.Google Scholar
  2. Butenandt, A. & P. Karlson, 1954. Über die isolierung eines metamorphose-hormons der insekten in kristallisierter form. Z. Naturforsch. 9B: 389–391.Google Scholar
  3. Chang, E. S., 1989. Endocrine regulation of molting in Crustacea. Rev. Aquat. Sci 1: 131–157.Google Scholar
  4. Chang, E. S. & J. D. O’Connor, 1979. Arthropod molting hormones. In Jaffe, B. M. & H. R. Behrman (eds), Methods of Hormone Radioimmunoassay, 2nd edn. Academic Press, New York: 797–814.Google Scholar
  5. Dennis, R., 1977. Partial characterization of and the effect of insect growth hormones on the ribosomes and polyribosomes of the nematode, Panagrellus redivivus. Int. J. Parasitol. 7: 171–179.Google Scholar
  6. Evershed, R. P., J. G. Mercer & H. H. Rees, 1987. Capillary gas chromatographymass spectrometry of ecdysteroids. J. Chromatogr. 390: 357–369.Google Scholar
  7. Evershed, R. P., M. Kabbouh, M. C. Prescott, J. L. Maggs & H. H. Rees, 1990. Current status and recent advances in the chromatography and mass spectrometry of ecdysteroids. In McCaffery, A. R. & I. D. Wilson (eds), Chromatography and Isolation of Insect Hormones and Pheromones. Plenum Press, New York: 103–115.Google Scholar
  8. Fleming, M. W., 1985. Ascaris suum: Role of ecdysteroids in molting. Exp. Parasitol. 60: 207–210.Google Scholar
  9. Fleming, M. W., 1987. Ecdysteroids during embryonation of eggs of Ascaris suum. Comp. Biochem. Physiol. 87A: 803–805.Google Scholar
  10. Fleming, M. W., 1993. Ecdysteroids during development in the ovine parasitic nematode, Haemonchus contortus. Comp. Biochem. Physiol. 104B: 653–655.Google Scholar
  11. Garcia, M., J. Gharbi, J-P. Girault, C. Hetru & R. Lafont, 1989. Ecdysteroid metabolism in leeches. Invert. Reprod. Dev. 15: 57–68.Google Scholar
  12. Gupta, A. P. (ed.), 1990a. Morphogenetic Hormones of Arthropods, 1. Discoveries, Syntheses, Metabolism, Evolution, Modes of Action, and Techniques. Rutgers Univ. Press, New Brunswick (NJ), 531 pp.Google Scholar
  13. Gupta, A. P. (ed.), 1990b. Morphogenetic Hormones of Arthropods, Volume 1.2. Embryonic and Postembryonic Sources. Rutgers Univ. Press, New Brunswick (NJ), 533 pp.Google Scholar
  14. Hampshire, F. & D. H. S. Horn, 1966. Structure of crustecdysone, a crustacean moulting hormone. J. Chem. Soc., Chem. Commun. 37–38.Google Scholar
  15. Horn, D. H. S., E. J. Middleton, J. A. Wunderlich & F. Hampshire, 1966. Identity of the moulting hormones of insects and crustaceans. J. Chem. Soc., Chem. Commun. 339–340.Google Scholar
  16. Horn, D. H. S. & R. Bergamasco, 1985. Chemistry of ecdysteroids. In Kerkut, G. A. & L. I. Gilbert (eds), Comprehensive Insect Physiology, Biochemistry, and Pharmacology, Volume 7. Pergamon Press, New York: 185–248.Google Scholar
  17. Horn, D. H. S., J. S. Wilkie & J. A. Thompson, 1974. Isolation of β-ecdysone (20-hydroxyecdysone) from the parasitic nematode Ascaris lumbricoides. Experientia 30: 1109–1110.Google Scholar
  18. Huber, R. & W. Hoppe, 1965. Zur Chemie des Ecdysons. VII. Die kristall-und Molekulstruktur Analyse des Insektenverpuppung-shormons Ecdyson mit der automatisierten Faltmolekulmethode. Chem. Ber. 98: 2403–2424.Google Scholar
  19. Kaplanis, J. N., W. E. Robbins, M. J. Thompson & S. R. Dutky, 1973. 26-Hydroxyecdysone: new insect moulting hormone from the egg of the tobacco hornworm. Science 180: 307–308.Google Scholar
  20. Karlson, P., H. Hoffmeister, H. Hummel, P. Hocks & G. Spiteller, 1965. Zur Chemie des Ecdysons. VI. Reaktionen des Ecdysonmolekuls. Chem. Ber. 98: 2394–2402.Google Scholar
  21. Mercer, J. G., J. P. Gillespie & H. H. Rees, 1988. The fate of [3H] ecdysone in three species of annelids. Int. J. Invert. Reprod. Dev. 13: 225–238.Google Scholar
  22. Morgan, E. D. & M. P. Marco, 1990. Advances in techniques for ecdysteroid analysis. Invert. Reprod. Dev. 18: 55–66.Google Scholar
  23. Okazaki, R. K., M. J. Snyder & E. S. Chang, 1988. Ecdysteroids in nemerteans: Presence and physiological role. Hydrobiologia 156: 153–160.Google Scholar
  24. Porchet, M., N. Gaillet, F. Sauber, M. Charlet & J. A. Hoffmann, 1984. Ecdysteroids in annelids. In Hoffmann, J. & M. Porchet (eds), Biosynthesis, Metabolism and Mode of Action of Invertebrate Hormones. Springer-Verlag, Berlin: 346–348.Google Scholar
  25. Snyder, M. J., R. K. Okazaki & E. S. Chang, 1992. Nemertean ecdysteroids: Relationship to reproduction. Invert. Reprod. Dev. 21: 7–13.Google Scholar
  26. Soumoff, C., D. H. S. Horn & J. D. O’Connor, 1981. Production of a new antiserum to arthropod molting hormone and comparison with two other antisera. J. Steroid Biochem. 14: 429–435.Google Scholar
  27. Thompson, M. J., J. N. Kaplanis, W. E. Robbins & R. T. Yamamoto, 1967. 20,26-Dihydroxyecdysone a new steroid with moulting hormone activity from the tobacco hornworm, Manduca sexta (Johannson). J. Chem. Soc., Chem. Commun. 650–653.Google Scholar

Copyright information

© Kluwer Academic Publishers 1997

Authors and Affiliations

  • Robert K. Okazaki
    • 1
  • Mark J. Snyder
    • 2
  • Casey C. Grimm
    • 3
  • Ernest S. Chang
    • 2
  1. 1.Department of Biological SciencesSoutheastern Louisiana UniversityHammondU.S.A
  2. 2.Bodega Marine LaboratoryUniversity of CaliforniaBodega BayU.S.A
  3. 3.Southern Regional Research Center, Agricultural Research ServiceUnited States Department of AgricultureNew OrleansU.S.A

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