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Two pupariation factors, anterior retraction factor (ARF) and puparium tanning factor (PTF), are absent from the hemolymph of larvae at the time of tanning accelerated by ARF/PTF, cyclic AMP, or dopamine. ARF and PTF are not involved in derepression of dopa decarboxylase (aromatic L-amino-acid decarboxylase, aromatic L-amino-acid carboxy-lyase, EC 4.1.1.28) synthesis initiated by ecdysone. Tanning is entirely inhibited by injection of two transcriptional inhibitors, actinomycin and BrdUrd, and two translational inhibitors, puromycin and cycloheximide. Retraction activity is more severely inhibited by the transcriptional than by the translational inhibitors. A tanning response is initiated by cyclic AMP in the presence of the transcriptional but not the translational inhibitors. Dihydric tanning substances (dopa, dopamine) initiate tanning in the presence of both types of inhibitors. Release of ARF and PTF from the central nervous system is inhibited by the four inhibitors. ARF totally reverses the inhibitory effects on retraction, whereas PTF does not reverse inhibition of tanning. These data are interpreted to mean that PTF is concerned with the regulation of two components of the tanning response: (i) acceleration of synthesis of a particular protein (associated with the tyrosine hydroxylation complex), and (ii) activation via cyclic AMP of a component of the tyrosine hydroxylating system.

Role of ecdysone, pupariation factors, and cyclic AMP in formation and tanning of the puparium of the fleshfly Sarcophaga bullata