Chitin biosynthesis in imaginal discs cultured in vitro

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Abstract
We have investigated the stimulation of cuticle production by imaginal discs ofPlodia interpunctella in tissue culture. We turned to biochemical methods to assess the quantitative effects of beta-ecdysone on chitin biosynthesis in wing discs incubated with 0.5 μC of C14-glucosamine for the final 24 h of culture. We demonstrated that imaginal discs ofP. interpunctella respond to increasing concentrations of β-ecdysone with increased synthesis. The threshold is between 0.01 and 0.1 μg/ml of hormone (2×10−8 M to 2×10−7 M). These data represent the first demonstration of quantitative biosynthesis of chitin by a developing tissue in vitro in relation to varying amounts of hormone. Additionally, protein synthesis during the β-ecdysone-dependent period was necessary for chitin synthesis. This system thus lends itself to a detailed investigation of the control of chitin biosynthesis.