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Open Access Research article

Global identification of bursicon-regulated genes in Drosophila melanogaster

Shiheng An1, Songjie Wang1, Lawrence I Gilbert2, Brenda Beerntsen3, Mark Ellersieck4 and Qisheng Song1*

Author Affiliations

1 Division of Plant Sciences, University of Missouri, Columbia, MO 65211, USA

2 Department of Biology, University of North Carolina, Chapel Hill, NC 27599-3280, USA

3 Department of Veterinary Pathobiology, University of Missouri, Columbia, MO 65211, USA

4 Experiment Station Statistics, University of Missouri, Columbia, MO 65211, USA

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BMC Genomics 2008, 9:424  doi:10.1186/1471-2164-9-424

Published: 18 September 2008

Abstract

Background

Bursicon is a heterodimer neuropeptide responsible for regulating cuticle sclerotization and wing expansion in several insect species. Recent studies indicate that the action of bursicon is mediated by a specific G protein-coupled receptor DLGR2 and the cAMP/PKA signaling pathway. However, little is known regarding the genes that are regulated by bursicon. The identification of bursicon-regulated genes is the focus of this investigation.

Results

We used DNA microarray analysis to identify bursicon-regulated genes in neck-ligated flies (Drosophila melanogaster) that received recombinant bursicon (r-bursicon). Fifty four genes were found to be regulated by bursicon 1 h post r-bursicon injection, 52 being up-regulated and 2 down-regulated while 33 genes were influenced by r-bursicon 3 h post-injection (24 up-regulated and 9 down-regulated genes). Analysis of these genes by inference from the fly database http://flybase.bio.indiana.edu webcite revealed that these genes encode proteins with diverse functions, including cell signaling, gene transcription, DNA/RNA binding, ion trafficking, proteolysis-peptidolysis, metabolism, cytoskeleton formation, immune response and cell-adhesion. Twenty eight genes randomly selected from the microarray-identified list were verified by real time PCR (qPCR) which supported the microarray data. Temporal response studies of 13 identified and verified genes by qPCR revealed that the temporal expression patterns of these genes are consistent with the microarray data.

Conclusion

Using r-bursicon, we identified 87 genes that are regulated by bursicon, 30 of which have no previously known function. Most importantly, all genes randomly selected from the microarray-identified list were verified by real time PCR. Temporal analysis of 13 verified genes revealed that the expression of these genes was indeed induced by bursicon and correlated well with the cuticle sclerotization process. The composite data suggest that these genes play important roles in regulating the cuticle sclerotization and wing expansion processes. The data obtained here will form the basis for future studies aimed at elucidating the exact mechanisms upstream from the secretion of bursicon and its binding to target cells.