The formation of the pupal cuticle by Drosophila imaginal discs in vitro

  • Fristrom J
  • Doctor J
  • Fristrom D
 et al. 
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Abstract

Mass-isolated imaginal discs of Drosophila melanogaster form a chitin-containing pupal procuticle In vitro. Optimal procuticle deposition occurs when the discs are incubated for 4-6 hr with 0.5-1.0 μg/ml of 20-hydroxyecdysone and then with less than 0.05 μg/ml of 20-hydroxyecdysone. The formation of the chitin-containing procuticle is demonstrated using three independent assays: with fluorescene-conjugated cuticle proteins that bind to chitin; by electron microscopy; by incorporation of [3H]glucosamine into a chitin fraction. Synthesis and deposition of pupal cuticle proteins are also demonstrated. Incorporation of [3H]glucosamine into chitin is sensitive to inhibitors of protein, RNA and chitin synthesis, but has little sensitivity to inhibitors of DNA synthesis, and dolichol-dependent glycosylation. © 1982.

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Authors

  • James W. Fristrom

  • John Doctor

  • Dianne K. Fristrom

  • W. Robert Logan

  • Donald J. Silvert

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