Session: SUN 203-235-Steroid Hormone Actions, Biosynthesis and Metabolism (posters)
Bench to Bedside
Poster Board SUN 235
Our previous studies using transgenic usp expressed under the control of the natural usp promoter in animals that were otherwise null for usp found that mutation to the ligand pocket (Q288A L366A), strongly reduced MF binding and blocked generation of the puparial cuticle.2,3 Presently, our microscopy studies determined that the epidermal sheet also prematurely separates from the cuticle, and the subsequent pupal stage animals rapidly dehydrate. Microarray expression analysis indicated that the majority of the most misexpressed (underexpressed) genes at the outset of processes leading to puparium formation are genes encoding cuticular structural proteins. qPCR analysis on isolated integument from that developmental stage confirmed the distinct misexpression of these genes. Expression analysis also confirmed that potential direct targets of MF/USP signaling (DHR3 (= ROR) and grainyhead) are misexpressed in the mutant QLUSP integument several hours before control animals pupariate. Chromatin immunoprecipitation, here validated by qPCR, confirmed that at this developmental period USP occupies a binding site in the DHR3 first intron.
Our results support the existence of a ligand/RXR (MF/USP) regulatory network for generation of a particular developmentally-timed and uniquely comprised epidermal cuticular barrier.
1Mace KA, Pearson JC, McGinnis W. Science. (2005)308: 381-5
2Jones D, Jones G, Teal PE. (2013) Gen Comp Endocrinol. 194: 326-335
3Jones G, Teal P, Henrich VC, Krzywonos A, Sapa A, Wozniak M, Smolka J, Jones D. (2013) Gen Comp Endocrinol.182:73-82
Nothing to Disclose: DJ, AN, AS, CC, RS, GJ
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