Utilization of Imaginal Tissues from Pupae of the Stable Fly for the Study of Chitin Synthesis and Screening of Chitin Synthesis Inhihitors12

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Abstract
Imaginal epidermal tissues from 4-day-old pupae of Stomoxys calcitrans (L.) were used to develop an assay suitable for studying chitin synthesis and for screening chitin synthesis inhibitors. Four chitin precursors, D-glucose, D-glucosamine, D-fructose, and N-acetyl-D-glucosamine were found to be suitable substrates in this assay system. All these substrates were incorporated into chitin. Maximum incorporation occurred within 8 h or less. The apparent I50’s of diflubenzuron (Dimilin; TH-6040; N-[[(4-chlorophenyl)amino]carbonyl]-2,6-difluorobenzamide), Bay Sir 8514 (2-chloro-N-[[[4(trifluoromethoxy) phenyl] amino] carbonyl] benzamide), EL-494 (N-[[[5-(4-bromophenyl)-6-methyl-2-pyrazinyl]-amino]carbonyl]-2,6-dichlorobenzamide), and polyoxin D for inhibition of NAGA incorporation were found to be 52, 440, 8,600 and 13,000 nM, respectively.
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