Ecdysteroid-mediated expression of hexamerin (arylphorin) in the rice moth, Corcyra cephalonica

Abstract

The insect development is intricately controlled by morphogenetic hormones, juvenile hormone (JH) and 20-hydroxyecdysone (20E) through the regulation of gene/protein expression. The role of hexamerins in the metamorphosis of insects and reproduction and their control by 20E at the gene level has been widely reported in insects. In the present study we for the first time report the role of ecdysteroids in the regulation of hexamerin synthesis in a lepidopteran insect Corcyra cephalonica. The hormonal studies were carried out using the normal and the thorax-ligated insects with both 20E and its non-steroidal agonist RH-5992. The in vitro as well as in vivo studies showed a stimulatory effect of 20E and its agonist on the hexamerin synthesis including arylphorin (Hex 2), whereas hormone blockade with azadirachtin caused a time dependent reduction in synthesis. The northern analysis using Hex 2b cDNA as probe too confirmed the above result. This was followed by the cloning of the Hex 2b gene. The full length of the genomic clone was found to be 3.5 kb long and has four exons interspersed by three introns. The genome walking analysis revealed the presence of a steroid hormone binding sequence "Ecdysone response element" (EcRE) in the 5' untranscribed region (UTR) of the gene. The data presented in this paper clearly suggest that hexamerin synthesis in C. cephalonica is transcriptionally regulated by 20E.