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. 1985 Jul 1;101(1):189–200. doi: 10.1083/jcb.101.1.189

The pupal cuticle of Drosophila: biphasic synthesis of pupal cuticle proteins in vivo and in vitro in response to 20-hydroxyecdysone

PMCID: PMC2113631  PMID: 3891759

Abstract

We investigated the synthesis and localization of Drosophila pupal cuticle proteins by immunochemical techniques using both a complex antiserum and monoclonal antibodies. A set of low molecular weight (15,000-25,000) pupal cuticle proteins are synthesized by the imaginal disk epithelium before pupation. After pupation, synthesis of the low molecular weight proteins ceases and a set of unrelated high molecular weight proteins (40,000-82,000) are synthesized and incorporated into the pupal cuticle. Ultrastructural changes in the cuticle deposited before and after pupation correlate with the switch in cuticle protein synthesis. A similar biphasic accumulation of low and high molecular weight pupal cuticle proteins is also seen in imaginal discs cultured in vitro. The low molecular weight pupal cuticle proteins accumulate in response to a pulse of the insect steroid hormone 20-hydroxyecdysone and begin to appear 6 h after the withdrawal of the hormone from the culture medium. The high molecular weight pupal cuticle proteins accumulate later in culture; a second pulse of hormone appears to be necessary for the accumulation of two of these proteins.

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Selected References

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  1. Chihara C. J., Silvert D. J., Fristrom J. W. The cuticle proteins of Drosophila melanogaster: stage specificity. Dev Biol. 1982 Feb;89(2):379–388. doi: 10.1016/0012-1606(82)90326-8. [DOI] [PubMed] [Google Scholar]
  2. Clarke L., Hitzeman R., Carbon J. Selection of specific clones from colony banks by screening with radioactive antibody. Methods Enzymol. 1979;68:436–442. doi: 10.1016/0076-6879(79)68033-3. [DOI] [PubMed] [Google Scholar]
  3. Fairbanks G., Steck T. L., Wallach D. F. Electrophoretic analysis of the major polypeptides of the human erythrocyte membrane. Biochemistry. 1971 Jun 22;10(13):2606–2617. doi: 10.1021/bi00789a030. [DOI] [PubMed] [Google Scholar]
  4. Fristrom J. W., Doctor J., Fristrom D. K., Logan W. R., Silvert D. J. The formation of the pupal cuticle by Drosophila imaginal discs in vitro. Dev Biol. 1982 Jun;91(2):337–350. doi: 10.1016/0012-1606(82)90040-9. [DOI] [PubMed] [Google Scholar]
  5. Fristrom J. W., Hill R. J., Watt F. The procuticle of Drosophila: heterogeneity of urea-soluble proteins. Biochemistry. 1978 Sep 19;17(19):3917–3930. doi: 10.1021/bi00612a005. [DOI] [PubMed] [Google Scholar]
  6. Fristrom J. W., Logan W. R., Murphy C. The synthetic and minimal culture requirements for evagination of imaginal discs of Drosophila melanogaster in vitro. Dev Biol. 1973 Aug;33(2):441–456. doi: 10.1016/0012-1606(73)90149-8. [DOI] [PubMed] [Google Scholar]
  7. Handler A. M. Ecdysteroid titers during pupal and adult development in Drosophila melanogaster. Dev Biol. 1982 Sep;93(1):73–82. doi: 10.1016/0012-1606(82)90240-8. [DOI] [PubMed] [Google Scholar]
  8. Hartree E. F. Determination of protein: a modification of the Lowry method that gives a linear photometric response. Anal Biochem. 1972 Aug;48(2):422–427. doi: 10.1016/0003-2697(72)90094-2. [DOI] [PubMed] [Google Scholar]
  9. Hawkes R., Niday E., Gordon J. A dot-immunobinding assay for monoclonal and other antibodies. Anal Biochem. 1982 Jan 1;119(1):142–147. doi: 10.1016/0003-2697(82)90677-7. [DOI] [PubMed] [Google Scholar]
  10. Hodgetts R. B., Sage B., O'Connor J. D. Ecdysone titers during postembryonic development of Drosophila melanogaster. Dev Biol. 1977 Oct 1;60(1):310–317. doi: 10.1016/0012-1606(77)90128-2. [DOI] [PubMed] [Google Scholar]
  11. Kessler S. W. Use of protein A-bearing staphylococci for the immunoprecipitation and isolation of antigens from cells. Methods Enzymol. 1981;73(Pt B):442–459. doi: 10.1016/0076-6879(81)73084-2. [DOI] [PubMed] [Google Scholar]
  12. Locke M., Huie P. Apolysis and the turnover of plasma membrane plaques during cuticle formation in an insect. Tissue Cell. 1979;11(2):277–291. doi: 10.1016/0040-8166(79)90042-9. [DOI] [PubMed] [Google Scholar]
  13. Marks E. P. Effects of ecdysterone on the deposition of cockroach cuticle in vitro. Biol Bull. 1972 Apr;142(2):293–301. doi: 10.2307/1540232. [DOI] [PubMed] [Google Scholar]
  14. Milner M. J. The eversion and differentiation of Drosophila melanogaster leg and wing imaginal discs cultured in vitro with an optimal concentration of beta-ecdysone. J Embryol Exp Morphol. 1977 Feb;37(1):105–117. [PubMed] [Google Scholar]
  15. Mitchell H. K., Petersen N. S. Rapid changes in gene expression in differentiating tissues of Drosophila. Dev Biol. 1981 Jul 15;85(1):233–242. doi: 10.1016/0012-1606(81)90253-0. [DOI] [PubMed] [Google Scholar]
  16. Mitsui T., Riddiford L. M. Pupal cuticle formation by Manduca sexta epidermis in vitro: patterns of ecdysone sensitivity. Dev Biol. 1976 Dec;54(2):172–186. doi: 10.1016/0012-1606(76)90297-9. [DOI] [PubMed] [Google Scholar]
  17. Nardi J. B., Willis J. H. Control of cuticle formation by wing imaginal discs in vitro. Dev Biol. 1979 Feb;68(2):381–395. doi: 10.1016/0012-1606(79)90212-4. [DOI] [PubMed] [Google Scholar]
  18. Richards G. Sequential gene activation by ecdysone in polytene chromosomes of Drosophila melanogaster. IV. The mid prepupal period. Dev Biol. 1976 Dec;54(2):256–263. doi: 10.1016/0012-1606(76)90303-1. [DOI] [PubMed] [Google Scholar]
  19. Richards G. Sequential gene activation by ecdysone in polytene chromosomes of Drosophila melanogaster. V. The late prepupal puffs. Dev Biol. 1976 Dec;54(2):264–275. doi: 10.1016/0012-1606(76)90304-3. [DOI] [PubMed] [Google Scholar]
  20. Richards G. The radioimmune assay of ecdysteroid titres in Drosophila melanogaster. Mol Cell Endocrinol. 1981 Mar;21(3):181–197. doi: 10.1016/0303-7207(81)90013-7. [DOI] [PubMed] [Google Scholar]
  21. Robb J. A. Maintenance of imaginal discs of Drosophila melanogaster in chemically defined media. J Cell Biol. 1969 Jun;41(3):876–885. doi: 10.1083/jcb.41.3.876. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. Roberts P. E., Willis J. H. The cuticular proteins of Tenebrio molitor. I. Electrophoretic banding patterns during postembryonic development. Dev Biol. 1980 Mar;75(1):59–69. doi: 10.1016/0012-1606(80)90143-8. [DOI] [PubMed] [Google Scholar]
  23. Roberts P. E., Willis J. H. The cuticular proteins of Tenebrio molitor. II. Patterns of synthesis during postembryonic development. Dev Biol. 1980 Mar;75(1):70–77. doi: 10.1016/0012-1606(80)90144-x. [DOI] [PubMed] [Google Scholar]
  24. Silvert D. J., Doctor J., Quesada L., Fristrom J. W. Pupal and larval cuticle proteins of Drosophila melanogaster. Biochemistry. 1984 Nov 20;23(24):5767–5774. doi: 10.1021/bi00319a015. [DOI] [PubMed] [Google Scholar]
  25. Snyder M., Hirsh J., Davidson N. The cuticle genes of drosophila: a developmentally regulated gene cluster. Cell. 1981 Jul;25(1):165–177. doi: 10.1016/0092-8674(81)90241-5. [DOI] [PubMed] [Google Scholar]
  26. Snyder M., Hunkapiller M., Yuen D., Silvert D., Fristrom J., Davidson N. Cuticle protein genes of Drosophila: structure, organization and evolution of four clustered genes. Cell. 1982 Jul;29(3):1027–1040. doi: 10.1016/0092-8674(82)90466-4. [DOI] [PubMed] [Google Scholar]
  27. de StGroth S. F., Scheidegger D. Production of monoclonal antibodies: strategy and tactics. J Immunol Methods. 1980;35(1-2):1–21. doi: 10.1016/0022-1759(80)90146-5. [DOI] [PubMed] [Google Scholar]

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