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Molecular and cDNA microarray analysis of steroid - induced muscle degeneration during insect metamorphosis

, : Molecular and cDNA microarray analysis of steroid - induced muscle degeneration during insect metamorphosis. Society for Neuroscience Abstract Viewer & Itinerary Planner : Abstract No 825 4

During metamorphosis in the hawkmoth, Manduca sexta, segmental homologs of the accessory planta retractor muscle (APRM) undergo programmed cell death (PCD) in a segment-specific pattern. The PCD of APRMs is triggered directly and cell-autonomously by the steroid hormone, 20-hydroxyecdysone (20E). To seek genes involved in intrinsic segmental identity, 20E signaling and muscle degeneration, we performed suppressive subtractive hybridization (SSH) on mRNA from APRMs in abdominal segments A3 and A6; APRM(6)s undergo PCD while APRM(3)s survive during the larval-pupal transformation. mRNA was obtained on the first day of the prepupal peak of 20E. We generated a library of approximately 2000 clones, some of which are likely redundant. These cDNAs, plus other candidate cDNAs, were spotted on microarrays and hybridizations are in progress to study gene expression in APRMs during prepupal 20E signaling. Five hundred of the clones were sequenced and, based on comparison with Drosophila and other species, many encode potentially interesting regulators of development. Concurrently, we examined the expression of genes in the Bithorax Complex (Bx-C) in APRMs, using real-time PCR. These genes regulate axial patterning of the insect abdomen. Real-time PCR data suggest that Bx-C genes are expressed in APRMs in a segmental pattern similar to that seen in fly embryos. The Bx-C genes appear to be expressed at very low levels in larvae but strongly upregulated by the prepupal peak of 20E. Potentially, the expression of axial patterning genes of the embryonic stage is recapitulated during metamorphosis to direct the segmental fates of individual cells in response to hormonal cues.


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