AbstractBursicon is a neuropeptide that regulates cuticle sclerotization (hardening and tanning) via a G protein–coupled receptor. It consists of two subunits, an alpha and a beta. In the present study, we investigated the transcriptional expression and in situ localization of bursicon α and β in the central nerve system of the house fly Musca domestica. Most importantly, we identified two novel bursicon‐regulated genes using recombinant bursicon (rbursicon) heterodimer in a neck‐ligated house fly assay. RT‐PCR analysis revealed that both bursicon α and β subunits were present in the central nerve system of larval and pupal stages, reached the maximal level in pharate adults, and declined sharply after adult emergence, suggesting the release of the hormone upon adult emergence. In situ localization of bursicon transcripts showed that both bursicon α and β transcripts were expressed in a set of neurosecretory cells (NSCs) in the thoracic‐abdominal ganglia of M. domestica. Two Drosophila melanogaster homologous genes, designated CG7985hh and CG30287hh, were up‐regulated by rbursicon in a time‐dependent manner and verified by real‐time PCR, implying their involvement in the cuticle tanning process. Arch. Insect Biochem. Physiol. 2008. © 2008 Wiley‐Liss, Inc.