The silkmoth homolog of the Drosophila ecdysone receptor (BI Isoform): Cloning and analysis of expression during follicular cell differentiation

Abstract

To understand the role that 20-hydroxy-ecdysone (20E) plays during ovarian development in Bombyx mori, we have undertaken the cloning of the silkworm ecdysone receptor (EcR) and a study of its expression during follicular cell differentiation. We have cloned a cDNA that contains a complete open reading frame for a 68.1 kDa polypeptide that shares extensive similarities with the B1 isoform of the Drosophila EcR. The presumed silkmoth EcR (BmEcR) is encoded by a single copy gene whose length is in excess of 23 kb. A portion of this gene encompassing seven exons that constitute the cloned BmEcR cDNA was also characterized. Employment of monoclonal antibodies, directed against the DNA binding domain of the Drosophila EcR, in Western blot analyses revealed the presence of a major 70 kDa polypeptide in extracts of follicular cells and other silkmoth tissues. The mRNA and protein encoded by BmEcR are present in constant amounts in follicular cells throughout vitellogenesis but disappear transiently at the onset of choriogenesis and reappear during the later stages of choriogenesis. The down-regulation of BmEcR in follicular cells during oogenesis suggests a complex relationship between 20E, the induction of the program of chorion gene expression in follicular cells during mid-vitellogenesis and the execution of this program at the end of vitellogenesis.