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Comparing in vivo and in vitro approaches to study the hormonal regulation of sea urchin reproduction

Published online by Cambridge University Press:  12 January 2016

Silvia Mercurio  and
Michela Sugni
Silvia Mercurio*
Affiliation:
Department of Biosciences, University of Milan, via Celoria, 26-20133 Milan, Italy
Michela Sugni
Affiliation:
Department of Biosciences, University of Milan, via Celoria, 26-20133 Milan, Italy
*
Correspondence should be addressed to:S. Mercurio, Department of Biosciences, University of Milan, via Celoria, 26-20133 Milan, Italy email: silvia.mercurio@unimi.it
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Abstract

Although in vivo and in vitro approaches appear to be very different, they are related and complementary techniques and both are essential for the investigation of diverse biological topics. The employment of both techniques was considered particularly appropriate to investigate the role of 17β-oestradiol and testosterone in echinoid reproductive biology. The relationship between sex-steroids and echinoid reproduction has not been clearly determined yet, due to the highly variable and sometimes contrasting results obtained from steroid administration experiments. These might be due to the activation of protective metabolic mechanisms that can prevent the exogenous molecules from exerting their biological functions, as observed in our previous research. To clarify these aspects, in the present study we explored sex-steroid involvement in the reproduction of the sea urchin Paracentrotus lividus, employing both in vivo and in vitro approaches: (1) an experiment involving hormone dietary administration was performed and different reproductive parameters were deeply analysed; (2) ovarian cells were cultured in the presence of the same steroids and morphological and biochemical analyses were carried out. According to our results, sex-steroids appear not to be involved in sea urchin gonad development and gamete maturation, as neither in vivo administration nor in vitro exposure influenced gonad and gamete growth. In addition, in vitro hormonal treatment did not affect sea urchin Major Yolk Protein content. Overall, the present work complements our previous research providing information on sex-steroid involvement in echinoid reproduction and illustrates new methodological approaches that will be useful for future research on invertebrate biology and physiology.

Keywords

steroid hormonesechinodermscell culturedietary administrationsea urchinreproduction
Type
Research Article
Information
Journal of the Marine Biological Association of the United Kingdom , Volume 96 , Issue 6 , September 2016 , pp. 1363 - 1372
Copyright
Copyright © Marine Biological Association of the United Kingdom 2016 

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References

REFERENCES

Barbaglio, A., Sugni, M., Di Benedetto, C., Bonasoro, F., Schnell, S., Lavado, R., Porte, C. and Candia Carnevali, D.M. (2007) Gametogenesis correlated with steroid levels during the gonadal cycle of the sea urchin Paracentrotus lividus (Echinodermata: Echinoidea). Comparative Biochemistry and Physiology Part A: Molecular and Integrative Physiology 147, 466–474.Google Scholar
Barker, M.F. and Xu, R.A. (1993) Effects of estrogens on gametogenesis and steroid levels in the ovaries and pyloric caeca of Sclerasterias mollis (Echinodermata: Asteroidea). Invertebrate Reproduction and Devevelopment 24, 53–58.Google Scholar
Creange, J.E. and Szego, C.M. (1967) Sulphation as a metabolic pathway for oestradiol in the sea urchin Strongylocentrotus franciscanus . Biochemical Journal 102, 898–904.CrossRefGoogle ScholarPubMed
Fujiwara, A., Unuma, T., Ohno, K. and Yamano, K. (2010) Molecular characterization of the major yolk protein of the Japanese common sea cucumber (Apostichopus japonicus) and its expression profile during ovarian development. Comparative Biochemistry and Physiology Part A: Molecular and Integrative Physiology 155, 34–40.CrossRefGoogle ScholarPubMed
Goldstone, J.V., Hamdoun, A., Cole, B.J., Howard-Ashby, M., Nebert, D.W., Scally, M., Dean, M., Epel, D., Hahn, M.E. and Stegeman, J.J. (2006) The chemical defensome: environmental sensing and response genes in the Strongylocentrotus purpuratus genome. Developmental Biology 300, 366–384.Google Scholar
Harrington, F.E. and Ozaki, H. (1986) The effect of estrogen on protein synthesis in echinoid coelomocytes. Comparative Biochemistry and Physiology Part B: Biochemistry and Molecular Biology 84, 417–421.Google Scholar
Hines, G.A., Watts, S.A. and McClintock, J.B. (1994) Biosynthesis of estrogen derivatives in the echinoid Lytechinus variegatus Lamarck. In David, G., Féral, J.-P. and Roux, M (eds) Echinoderms through time. Rotterdam: Balkema, pp. 711–715.Google Scholar
Hochberg, R.B. (1998) Biological esterification of steroids. Endocrine Reviews 19, 331–348.Google Scholar
Janer, G., LeBlanc, G.A. and Porte, C. (2005a) A comparative study on androgen metabolism in three invertebrate species. General and Comparative Endocrinology 143, 211–221.Google Scholar
Janer, G., Sternberg, R.M., LeBlanc, G.A. and Porte, C. (2005b) Testosterone conjugating activities in invertebrates: are they targets for endocrine disruptors? Aquatic Toxicology 71, 273–282.Google Scholar
Köhler, H-R., Kloas, W., Schirling, M., Lutz, I., Reye, A., Langen, J.-S., Triebskorn, R., Nagel, R. and Schönfelder, G. (2007) Sex steroid receptor evolution and signalling in aquatic invertebrates. Ecotoxicology 16, 131–143.CrossRefGoogle ScholarPubMed
Kontrogianni-Konstantopoulos, A. and Flytzanis, C.N. (2001) Differential cellular compartmentalization of the nuclear receptor SpSHR2 splicing variants in early sea urchin embryos. Molecular Reproduction and Development 60, 147–157.Google Scholar
Kontrogianni-Konstantopoulos, A., Leahy, P.S. and Flytzanis, C.N. (1998) Embryonic and post-embryonic utilization and subcellular localization of the nuclear receptor SpSHR2 in the sea urchin. Journal of Cell Science 111, 2159–2169.Google Scholar
Kontrogianni-Konstantopoulos, A., Vlahou, A., Vu, D. and Flytzanis, C.N. (1996) A novel sea urchin nuclear receptor encoded by alternatively spliced maternal RNAs. Developmental Biology 177, 371–382.Google Scholar
Laemmli, U.K. (1970) Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature 227, 680–685.Google Scholar
Lange, I.G., Hartel, A. and Meyer, H.H.D. (2002) Evolution of oestrogen functions in vertebrates. Journal of Steroid Biochemistry 83, 219–226.Google Scholar
Lavado, R., Sugni, M., Candia Carnevali, M.D. and Porte, C. (2006) Triphenyltin alters androgen metabolism in the sea urchin Paracentrotus lividus . Aquatic Toxicology 79, 247–256.Google Scholar
Mercurio, S., Di Benedetto, C., Sugni, M. and Candia Carnevali, M.D. (2014) Primary cell cultures from sea urchin ovaries: a new experimental tool. In Vitro Cellular and Developmental Biology – Animal 50, 139–145.Google Scholar
Mercurio, S., Tremolada, P., Nobile, M., Fernandes, D., Porte, C. and Sugni, M. (2015) Unraveling estradiol metabolism and involvement in the reproductive cycle of non-vertebrate animals: the sea urchin model. Steroids 104, 25–36.Google Scholar
Prowse, T.A. and Byrne, M. (2012) Evolution of yolk protein genes in the Echinodermata. Evolution and Development 14, 139–151.Google Scholar
Roepke, T.A., Chang, E.S. and Cherr, G.N. (2006) Maternal exposure to estradiol and endocrine disrupting compounds alters the sensitivity of sea urchin embryos and the expression of an orphan steroid receptor. Journal of Experimental Zoology A: Comparative Experimental Biology 305A, 830–841.Google Scholar
Roepke, T.A., Snyder, M.J. and Cherr, G.N. (2005) Estradiol and endocrine disrupting compounds adversely affect development of sea urchin embryos at environmentally relevant concentrations. Aquatic Toxicology 71, 155–173.Google Scholar
Schoenmakers, H.J.N., Van Bohemen, C.G. and Dieleman, S.J. (1981) Effects of oestradiol-17β on the ovaries of the starfish Asterias rubens . Development Growth and Differentiation 23, 125–135.Google Scholar
Shyu, A.B., Blumenthal, T. and Raff, R.A. (1987) A single gene encoding vitellogenin in the sea urchin Strongylocentrotus purpuratus: sequence at the 5′ end. Nucleic Acids Research 15, 10405–10417.Google Scholar
Shyu, A.B., Raff, R.A. and Blumenthal, T. (1986) Expression of the vitellogenin gene in female and male sea urchin. Proceedings of the National Academy of Sciences USA 83, 3865–3869.Google Scholar
Spirlet, C., Grosjean, P. and Jangoux, M. (2000) Optimization of gonad growth by manipulation of temperature and photoperiod in cultivated sea urchins, Paracentrotus lividus (Lamarck) (Echinodermata). Aquaculture 185, 85–99.Google Scholar
Strott, C.A. (1996) Steroid sulfotransferases. Endocrine Reviews 17, 670–697.Google Scholar
Sugni, M., Motta, D., Tremolada, P. and Candia Carnevali, M.D. (2012) Exploring endocrine regulation of sea urchin reproductive biology: effects of 17β-estradiol. Journal of the Marine Biological Association of the United Kingdom 92, 1419–1426.Google Scholar
Takahashi, N. and Kanatani, H. (1981) Effect of 17β-Estradiol on growth of oocytes in cultured ovarian fragments of the starfish, Asterina pectinifera . Development Growth and Differentiation 23, 565–569.Google Scholar
Unuma, T., Okamoto, H., Konishi, K., Ohta, H. and Mori, K. (2001) Cloning of cDNA encoding vitellogenin and its expression in red sea urchin, Pseudocentrotus depressus . Zoological Science 18, 559–565.Google Scholar
Unuma, T., Sawaguchi, S., Yamano, K. and Ohta, H. (2011) Accumulation of the major yolk protein and zinc in the agametogenic sea urchin gonad. Biological Bulletin 221, 227–237.Google Scholar
Unuma, T., Suzuki, T., Kurokawa, T., Yamamoto, T. and Akiyama, T. (1998) A protein identical to the yolk protein is stored in the testis in male red sea urchin, Pseudocentrotus depressus . Biological Bulletin 194, 92–97.Google Scholar
Unuma, T., Yamamoto, T. and Akiyama, T. (1999) Effect of steroids on gonadal growth and gametogenesis in the juvenile red sea urchin Pseudocentrotus depressus . Biological Bulletin 196, 199–204.Google Scholar
Unuma, T., Yamamoto, T., Akiyama, T., Shiraishi, M. and Ohta, H. (2003) Quantitative changes in yolk protein and other components in the ovary and testis of the sea urchin Pseudocentrotus depressus . Journal of Experimental Biology 206, 365–372.Google Scholar
Van der Plas, A.J., Koenderman, A.H.L., Deibel-van Schijndel, G.J. and Voogt, P.A. (1982) Effects of oestradiol-17β on the synthesis of RNA, proteins and lipids in the pyloric caeca of the female starfish Asterias rubens . Comparative Biochemistry and Physiology Part B: Biochemistry and Molecular Biology 73, 965–970.Google Scholar
Varaksina, G.S. and Varaksin, A.A. (2001) The effect of estradiol dipropionate on the rate of protein synthesis in the testicle of the sea urchin Strongylocentrotus nudus . Russian Journal of Marine Biology 27, 94–97.Google Scholar
Varaksina, G.S. and Varaksin, A.A. (2002) Effect of estradiol dipropionate on protein synthesis in oocytes and ovary of the sea urchin Strongylocentrotus intermedius at different stages of the reproductive cycle. Biological Bulletin 29, 496–500.Google Scholar
Voogt, P.A. and Van Rheenen, J.W.A. (1986) Androstenedione metabolism in the sea star Asterias rubens L. Studied in homogenates and intact tissue: biosynthesis of the novel steroid fatty-acyl testosterone. Comparative Biochemistry and Physiology Part B: Biochemistry and Molecular Biology 85, 497–501.Google Scholar
Waal, D.M., Portman, J. and Voogt, P.A. (1982) Steroid receptors in invertebrates. A specific 17β-estradiol binding protein in a sea star. Marine Biology Letters 3, 317–323.Google Scholar
Wasson, K.M., Gower, B.A., Hines, G.A. and Watts, S.A. (2000a) Levels of progesterone, testosterone, and estradiol, and androstenedione metabolism in the gonads of Lytechinus variegatus (Echinodermata:Echinoidea). Comparative Biochemistry and Physiology Part C: Toxicology and Pharmacology 126, 153–165.Google Scholar
Wasson, K.M., Gower, B.A. and Watts, S.A. (2000b) Responses of ovaries and testes of Lytechinus variegatus (Echinodermata: Echinoidea) to dietary administration of estradiol, progesterone and testosterone. Marine Biology 137, 245–255.Google Scholar