Advertisement

Acta Biologica Hungarica

, Volume 52, Issue 4, pp 457–471 | Cite as

Localization of a Cuticular Protein During the Postembryonal Development of Manduca sexta

  • Gy. CsikósEmail author
  • Kinga Molnár
  • Noémi H. Borhegyi
  • M. Sass
Article

Abstract

The pattern of cuticle protein synthesis by the epidermis of insects changes during the last larval, pupal and adult development, leading to an alteration in cuticular stucture and feature. We have isolated a protein that had an apparent molecular mass of 33.1 kD from larval cuticle of Manduca sexta. Synthesis, transport and accumulation of MsCP33.1 were followed during metamorphosis by immunoblots and immunocytochemical methods using the antibody developed against this protein. Our data prove that the presence of MsCP33.1 in the larval cuticle is general while its appearance in the pupal or adult integument is restricted only in the cuticle of wings and apodemes. We established that the synthesis of 33.1 kD protein is negatively regulated by moulting hormone (20-hydroxyecdysone). Possible roles for this cuticular protein are discussed.

Keywords

Cuticular protein immunolocalization moulting hormone metamorphosis 

Preview

Unable to display preview. Download preview PDF.

Unable to display preview. Download preview PDF.

References

  1. 1.
    Andersen, S. O., Rafn, K., Krogh, T. N., Hojrup, P., Roepstorff, P. (1995) Comparison of larval and pupal cuticular proteins in Tenebrio molitor. Insect Biochem. Molec. Biol. 25, 177–187.Google Scholar
  2. 2.
    Apple, R. T., Fristrom, J. W. (1991) 20-hydroxyecdysone is required for, and negatively regulates, transcription of Drosophila pupal cuticle protein genes. Dev. Biol. 146, 569–582.PubMedGoogle Scholar
  3. 3.
    Bell, R. A., Joachim, F. G. (1976) Techniques for rearing laboratory colonies of the tobacco hornworm and pink bollworms. Ann. Entomol. Soc. Am. 69, 365–373.Google Scholar
  4. 4.
    Binger, L. C., Willis, J. H. (1990) In vitro translation of epidermal RNA from different anatomical regions metamorphic stages. Insect Biochem. 20, 573–583.Google Scholar
  5. 5.
    Binger, L. S., Willis, J. H. (1994) Identification of the cDNA, gene and promoter for a major protein from flexible cuticles of the giant silkmoth Hyalophora cecropia. Insect Biochem. Molec. Biol. 24, 989–1000.Google Scholar
  6. 6.
    Bouhin, H., Charles, J.-P., Quennedey, B., Delachambre, J. (1992) Developmental profiles of epidermal mRNA content during the development of Bombyx mori larvae. Insect Biochem. 19, 29–39.Google Scholar
  7. 7.
    Brey, T. P., Lee, W. J., Yamakawa, M., Koizumi, Y., Perrot, S., Francois, M., Ashida, M. (1993) Role of the integument in insect immunity: Epicuticular abrasion and induction of cecropin synthesis in cuticular epithelial cells. Proc. Natl. Acad. Sci. USA. 90, 6275–6279.PubMedGoogle Scholar
  8. 8.
    Charles, J. -P, Bouhin, H., Quennedey, B., Current, A., Delachambre, J. (1992) CDNA cloning and deduced amino acid sequence of a major, glycine-rich cuticular protein from the coleopteran Tenebrio molitor. Temporal and spatial distribution of the transcript during metamorphosis. Eur. J. Biochem. 206, 813–819.PubMedGoogle Scholar
  9. 9.
    Cox, D. C., Willis, J. H. (1985) The cuticular proteins of Hyalophora cecropia from different anatomical regions and metamorphotic stages. Insect Biochem. 146, 349–362.Google Scholar
  10. 10.
    Csikós, Gy., Molnár, K., Borhegyi, H. N., Talián, Cs. G., Sass, M. (1999) Insect cuticle, an in vivo model of protein trafficking. J. Cell Science 112, 2113–2124.PubMedGoogle Scholar
  11. 11.
    Del Castillo, P., Llorente, A. R., Stockert, J. C. (1989) Influence of fixation, axciting light and section thickness on the primary fluorescence of samples for microfluorometric analysis. Basic Appl. Histochem. 33, 251–257.PubMedGoogle Scholar
  12. 12.
    Fechtel, K., Fristrom, D. K., Fristrom, J. W. (1989) Prepupal differentiation in Drosophila: distinct cell types elaborate a shared structure, the pupal cuticle, but accumulate transcriptsin unique patterns. Development 106, 649–656.PubMedGoogle Scholar
  13. 13.
    Hawkes, R. (1982) Identification of Concanavalin A binding sites after sodium dodecyl sulphate-gel electrophoresis and protein blotting. Anal. Biochem. 123, 143–146.PubMedGoogle Scholar
  14. 14.
    Hiruma, K., Hardie, J., Riddiford, L. M. (1991) Hormonal regulation of epidermal metamorphosis in vitro, control of expression of a larval-specific cuticle gene. Dev. Biol. 144, 369–378.PubMedGoogle Scholar
  15. 15.
    Hopkins, T. L., Krchma, L. J., Ahmad, S. A., Kramer, K. J. (2000) Pupal cuticle of Manduca sexta: characterization and profiles during sclerotization. Insect Biochem. Mol. Biol. 30, 19–27.PubMedGoogle Scholar
  16. 16.
    Horodyski, F. M., Riddiford, L. M. (1989) Expression and hormonal control of a new larval cuticular multigene family at the onset of metamorphosis of the tobacco hornworm. Dev. Biol. 132, 292–303.PubMedGoogle Scholar
  17. 17.
    Jensen, U. G., Rothman, A., Skou, L., Andersen, S. O., Roepstorff, P., Hojrup, P. (1997) Cuticular proteins from the giant cockroach, Blaberus cranifer. Insect Biochem. Molec. Biol. 27, 109–120.Google Scholar
  18. 18.
    Kiely, M. L., Riddiford, L. M. (1985) Temporal programming of epidermal cell protein synthesis during the larval-pupal transformation of Manduca sexta. Roux’s Arch. Dev. Biol. 194, 325–335.Google Scholar
  19. 19.
    Koeppe, J. K., Gilbert, L. I. (1973) Immunochemical evidence for the transport of hemolymph protein into the cuticle of Manduca sexta. J. Insect Phys. 19, 615–624.Google Scholar
  20. 20.
    Laemmli, U. K. (1970) Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature, 227, 680–685.PubMedGoogle Scholar
  21. 21.
    Lampe, J. D., Willis, J. H. (1994) Characterization of a cDNA and gene encoding a cuticular protein from rigid cuticles of the giant silkmoth Hyalophora cecropia. Insect Biochem. Molec. Biol. 24, 419–435.Google Scholar
  22. 22.
    Locke, M., Kiss, A., Sass, M. (1994) The cuticular localization of integument peptides from particular routing categories. Tissue Cell 26, 707–734.PubMedGoogle Scholar
  23. 23.
    Marmaras, V. J., Charalambidis, N. D., Zervas, Ch. G. (1996) Immune response in insects: the role of phenoloxydase in defense reactions in relation to melanization and sclerotization. Arch. Insect Biochem. Phys. 31, 119–133.Google Scholar
  24. 24.
    Missios, S., Davidson, H. C., Linder, D., Mortimer, L., Okobi, A. O., Doctor, J. S. (2000) Characterization of cuticular proteins in the red flour beetle, Tribolium castaneum. Insect Biochem. Mol. Biol. 30, 47–56.PubMedGoogle Scholar
  25. 25.
    Nakato, H., Izumi, S., Tamino, S. (1992) Structure and expression of gene coding for a pupal cuticle protein of Bombyx mori. Biochem, Biophys. Acta 1132, 161–167.Google Scholar
  26. 26.
    Palli, S. R., Locke, M. (1987) The synthesis of hemolymph proteins by the larval epidermis of an insect Calpodes ethlius (Lepidoptera: hesperidae). Insect Biochem. 17, 711–722.Google Scholar
  27. 27.
    Rebers, J., Riddiford, L. M. (1988) Structure and expression of a Manduca sexta larval cuticle gene homologous to Drosophila cuticle genes. J. Mol. Biol. 203, 411–423.PubMedGoogle Scholar
  28. 28.
    Riddiford, L. M. (1978) Ecdysone-induced change in cellular commitment of the epidermis of the tobacco hornworm Manduca sexta, at the initiation of metamorphosis. Gen. Comp. Endocrinol. 34, 438–446.PubMedGoogle Scholar
  29. 29.
    Riddiford, L. M., Baeckmann, A., Hice, R. H., Rebers, J. (1986) Developmental expression of three genes for larval cuticular proteins of the tobacco hornworm, Manduca sexta. Dev. Biol. 118, 82–94.PubMedGoogle Scholar
  30. 30.
    Samuels, R. I., Reynolds, S. E. (1993) Moulting fluid enzymes of the tobacco hornworm, Manduca sexta: timing of proteolytic and chinolytic activity in relation to pre-ecdysial development. Arch. Insect Phys. Biochem. 24, 33–44.Google Scholar
  31. 31.
    Sass, M., Kiss, A., Locke, M. (1993) Classes of integument peptides. Insect Biochem. Molec. Biol. 23, 845–857.Google Scholar
  32. 32.
    Sass, M., Kiss, A., Locke, M. (1994) The localization of surface integument peptides in tracheae and tracheoles. J. Insect Phys. 40, 561–575.Google Scholar
  33. 33.
    Sass, M., Kiss, A., Locke, M. (1994) Integument and hemocyte peptides. J. Insect Physiol. 40, 407–421.Google Scholar
  34. 34.
    Sass, M., Löw, P., Móricz, Zs., Csikós, G., Kovács, J. (1991) The fate and possible role of arylphoryn during the metamorphosis of Mamestra brassicae. Acta Biol. Hung. 42, 141–160.PubMedGoogle Scholar
  35. 35.
    Schenkel, H., Scheller, K. (1986) Stage and tissue specific expression of the genesencoding Calliphorin, the major larval serum protein of Calliphora vicina. Roux’s Archs. Dev. Biol. 195, 290–295.Google Scholar
  36. 36.
    Snyder, M., Hunkapiller, M., Yuen, D., Silvert, D., Fristrom, J., Davidson, N. (1982) Cuticle protein genes in Drosophila. Structure, organization and evolution of four clustered genes. Cell, 29, 1024–1040.Google Scholar
  37. 37.
    Stiles, B., Leopold, R. A. (1990) Cuticle proteins from the Anthonomus grandis abdomen: stage specificity and immunological relatedness. Insect Biochem. 20, 113–125.Google Scholar
  38. 38.
    Towbin, H., Staehelin, T., Gordon, J. (1979) Electrophoretic transfer of proteins from polyacrylamide gels to nitrocellulose sheets: procedure and some applications. Proc. Natl. Acad. Sci. USA 76, 4350–4354.PubMedGoogle Scholar
  39. 39.
    Wielgus, J. J. (1983) Stimulation of intermoult cuticle deposition by a hemolymph trophic factor of the tobacco hornworm, Manduca sexta. Insect Biochem. 13, 313–322.Google Scholar
  40. 40.
    Wielgus, J. J., Caldwell, G. A., Nichols, R. L., White, C. F. (1990) Purification, properties, and titer of hemolymph trophic factor in larvae and pupae of Manduca sexta. Insect Biochem. 20, 65–72.Google Scholar
  41. 41.
    Willis, J. H., Reiger, J. C., DeBrunner, B. A. (1981) The metamorphosis of arthropodin. In: Bhaskaran, G., Friedmann, S., Rodrigez, J. G. (eds) Current topics in insect endocrinology and nutrition. Plenum Press, New York, pp. 26–46.Google Scholar
  42. 42.
    Willis, J. H. (1996) Metamorphosis of the cuticle, its proteins, and their genes. In: Gilbert, L. I., Tate, J. R., Atkinson, B. G. (eds) Metamorphosis: Postembryonic reprogramming of gene expression in amphibian and insect cells. Academic Press, Inc., San Diego, pp. 254–282.Google Scholar
  43. 43.
    Wolfgang, W. J., Fristrom, D., Fristrom, J. W. (1986) The pupal cuticle of Drosophila: differential ultrastructural immunolocalization of cuticle proteins. J. Cell Biol. 102, 306–311.PubMedGoogle Scholar
  44. 44.
    Wolfgang, W. J., Riddiford, L. M. (1986) Larval cuticular morphogenesis in the tobacco hornworm, Manduca sexta, and its hormonal regulation. Dev. Biol. 113, 305–316.PubMedGoogle Scholar

Copyright information

© Akadémiai Kiadó, Budapest 2001

This article is distributed under the terms of the Creative Commons Attribution 4.0 International License (http://creativecommons.org/licenses/by/4.0/), which permits unrestricted use, distribution, and reproduction in any medium, provided you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made.

Authors and Affiliations

  • Gy. Csikós
    • 1
    Email author
  • Kinga Molnár
    • 1
  • Noémi H. Borhegyi
    • 1
  • M. Sass
    • 1
  1. 1.Department of General ZoologyEötvös Loránd UniversityBudapestHungary

Personalised recommendations