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Demonstration of cytoplasmic receptors for the molting hormones in crayfish

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Summary

From in vitro experiments using different binding assays it is in crayfish demonstrated that the cytosol of target tissues is able to bind both ecdysone and ecdysterone. The ability to bind ecdysteroids is destroyed by heating and by treatment with α-chymotrypsin and N-ethyl-maleinimide (NEM) (Figs. 4, 5). In target tissues there is a strong positive correlation between protein content and binding (Fig. 6). The association of the hormone-protein-complex is rapid, taking only a few min even at 5° C (Fig. 3). The binding of the two hormones to the cytosol is both specific and saturable. The association constants for the cytoplasmic receptors from hypodermis, hindgut and midgut gland are in the range of 3–6×107 M−1 for ecdysone and 5–7×108 M−1 for ecdysterone (Fig. 8). The data suggest the existence of cytoplasmic ecdysteroid receptors.

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References

  • Andrieux, N., Porcheron, P., Berreur-Bonnenfant, J., Dray, F.: Détermination du taux d'ecdysone au cours du cycle d'intermue chez le crabeCarcinus maenas; comparaison entre individus sains et parasités parSacculina carcini. C.R. Acad. Sci.283D, 1429–1432 (1976)

    Google Scholar 

  • Ashburner, M.: N-ethylmaleinimide inhibition of the induction of gene activity by the hormone ecdysone. FEBS Lett.22, 265–269 (1972)

    Google Scholar 

  • Beckers, C., Emmerich, H.: The ecdysone titer inDrosophila hydei and the metabolism of the moulting hormones in the salivary gland. Colloques internationaux C.N.R.S., No. 251 — Actualités sur les hormones d'invertébrés, pp. 351–357 (1976)

  • Best-Belpomme, M., Courgeon, A.-M.: Presence ou absence des récepteurs saturable de l'ecdystérone dans des clones sensibles ou résistants deDrosophila melanogaster en culture in vitro. C. R. Acad. Sci.280D, 1397–1400 (1975)

    Google Scholar 

  • Blanchet, M.-F., Porcheron, P., Dray, F.: Étude des variations du taux des ecdysones au cours du cycle d'intermue chez le male d'Orchestia gammarella Pallas (Crustacé Amphipode) par dosage radio-immunologique. C. R. Acad. Sci.283D, 651–654 (1976)

    Google Scholar 

  • Butterworth, F.M., Berendes, H.D.: Ecdysone-binding proteins in haemolymph and tissues ofDrosophila hydei larvae. J. Insect Physiol.20, 2195–2205 (1974)

    Google Scholar 

  • Chamness, G.C., McGuire, W.: Scatchard plots: Common errors in correction and interpretation. Steroids26, 538–542 (1975)

    Google Scholar 

  • Chang, E.S., O'Connor, J.D.: Secretion of α-ecdysone by crab Y-organs in vitro. Proc. Natl. Acad. Sci. USA74, 615–618 (1977)

    Google Scholar 

  • Chang, E.S., Sage, B.A., O'Connor, J.D.: The qualitative and quantitative determinations of ecdysones in tissues of the crab,Pachygrapsus crassipes, following molt induction. Gen. Comp. Endocrinol.30, 21–33 (1976)

    Google Scholar 

  • Charmantier, G., Ollé, M., Trilles, J.P.: Aspectes du dosage de l'ecdystérone chezSphaeroma serratum (Crustacea, Isopoda, Flabellifera) et premiers résultats. C. R. Acad. Sci.283D, 1329–1331 (1976)

    Google Scholar 

  • Emmerich, H.: Über die Hämolymphproteine vonPyrrhocoris apterus und über die Bindung von Ecdyson durch die Hämolymphproteine. J. Insect Physiol.16, 725–747 (1970a)

    Google Scholar 

  • Emmerich, H.: Ecdysonbindende Proteinfraktionen in den Speicheldrüsen vonDrosophila hydei. Z. vergl. Physiol.68, 385–402 (1970b)

    Google Scholar 

  • Emmerich, H.: Ecdysone binding proteins in nuclei and chromatin fromDrosophila salivary glands. Gen. Comp. Endocrinol.19, 543–551 (1972)

    Google Scholar 

  • Emmerich, H.: Über den Wirkungsmechanismus der Häutungshormone der Insekten. In: Verh. Dtsch. Zool. Ges. Erlangen. Rathmayer, W. (ed.), pp. 184–201. Stuttgart: Fischer 1977

    Google Scholar 

  • Feyereisen, R.: A specific binding protein for the moulting hormone ecdysterone in locust haemolymph. Experientia33, 1111–1113 (1977)

    Google Scholar 

  • Feyereisen, R., Lagueux, M., Hoffmann, J.A.: Dynamics of ecdysone metabolism after ingestion and injection inLocusta migratoria. Gen. Comp. Endocrinol.29, 319–327 (1976)

    Google Scholar 

  • Gilbert, L.I., King, D.S.: Physiology of growth and development: Endocrine aspects. In: The physiology of insecta, Vol. 1. Rockstein, M. (ed.), pp. 250–370. New York: Academic Press 1973

    Google Scholar 

  • Gorell, T.A., Gilbert, L.I., Siddall, J.B.: Binding proteins for an ecdysone metabolite in the crustacean hepatopancreas. Proc. Natl. Acad. Sci. USA69, 812–815 (1972)

    Google Scholar 

  • Harreveld, A. van: Physiological saline for crayfish. Proc. Soc. Exp. Biol. Med.34, 428–432 (1936)

    Google Scholar 

  • Jensen, E.V., DeSombre, E.R.: Mechanism of action of the female sex hormones. Ann. Rev. Biochem.41, 203–230 (1972)

    Google Scholar 

  • Kuppert, P., Büchler, M., Spindler, K.-D.: Distribution and transport of molting hormones in the crayfish,Orconectes limosus. Z. Naturforsch.33c, 437–441 (1978)

    Google Scholar 

  • Liao, S.: Cellular receptors and mechanism of action of steroid hormones. Int. Rev. Cytol.41, 87–172 (1975)

    Google Scholar 

  • Lowry, O.H., Rosebrough M.J., Farr, A.L., Randall, R.J.: Protein measurement with the Folin Phenol reagent. J. Biol. Chem.193, 265–275 (1951)

    Google Scholar 

  • Natori, S., Ohtaki, T.: Uptake of β-ecdysone by the fat body and membrane vesicles of fat body cells ofSarcophaga peregrina larvae. J. Insect Physiol.22 227–234 (1976)

    Google Scholar 

  • Rao, G.S., Schulze-Hagen, K., Rao, M.L., Breuer, H.: Kinetics of steroid transport through cell membranes: comparison of the uptake of cortisol by isolated rat liver cells with binding of cortisol to rat liver cytosol. J. Steroid Biochem.7, 1123–1129 (1976)

    Google Scholar 

  • Scatchard, G.: The attractions of proteins for small molecules and ions. Ann. NY Acad. Sci.51, 660–672 (1949)

    Google Scholar 

  • Schrader, W.T., Smith, R.G., Coty, W.A.: Steroid hormone receptor purification and characterization. In: Laboratory methods manual for hormone action and molecular endocrinology Schrader, W.T., O'Malley, B.W. (eds.), pp. 3.1–3.31. Houston, TX: Department of Cell Biology, Baylor College of Medicine, Texas Medical Center 1977

    Google Scholar 

  • Sekeris, C.E.: Molecular action of insect hormones. In: Invertebrate endocrinology and hormonal heterophylly. Burdette, W.J. (ed.), pp. 55–86. Berlin, Heidelberg, New York: Springer 1974

    Google Scholar 

  • Sheridan, P.J.: Is there an alternative to the cytoplasmic receptor model for the mechanism of action of steroids? Life Sci.17, 497–502 (1975)

    Google Scholar 

  • Sridhara, S., Nowock, J., Gilbert, L.I.: Biochemical endocrinology of insect growth and development. Int. Rev. Biochem.20, 133–188 (1978)

    Google Scholar 

  • Thamer, G., Karlson, P.: Nachweis der Proteinbindung von Ecdyson bei der SchmeißfliegeCalliphora erythrocephala. Z. Naturforsch.27b, 1191–1195 (1972)

    Google Scholar 

  • Willig, A.: Die Rolle der Ecdysone im Häutungszyklus der Crustaceen. Fortschr. Zool.22, 55–74 (1974)

    Google Scholar 

  • Willig, A., Keller, R.: Molting hormone content, cuticle growth and gastrolith growth in the molt cycle of the crayfishOrconectes limosus. J. comp. Physiol.86, 377–388 (1973)

    Google Scholar 

  • Willig, A., Keller, R.: Biosynthesis of α- and β-ecdysone by the crayfishOrconectes limosus in vivo and by its Y-organs in vitro. Experientia32, 936–937 (1976)

    Google Scholar 

  • Yamamota, K.R., Alberts, B.M.: Steroid receptors: Elements for modulation of eukaryotic transcription. Ann. Rev. Biochem.45, 721–746 (1976)

    Google Scholar 

  • Yund, M.A., Fristrom, J.W.: Uptake and binding of β-ecdysone in imaginal discs ofDrosophila melanogaster. Dev. Biol.43, 287–298 (1975a)

    Google Scholar 

  • Yund, M.A., Fristrom, J.W.: Specific binding of β-ecdysone to imaginal discs ofDrosophila melanogaster. In: Development biology, Vol. 2. McMahon, D., Fox, C.F. (eds.), pp. 404–408. Menlo Park, CA: Benjamin 1975b

    Google Scholar 

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Kuppert, P., Wilhelm, S. & Spindler, KD. Demonstration of cytoplasmic receptors for the molting hormones in crayfish. J Comp Physiol B 128, 95–100 (1978). https://doi.org/10.1007/BF00689472

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