title: Differential incorporation of cholesterol and cholesterol derivatives into ecdysteroids by the larval ring glands and adult ovaries of Drosophila melanogaster: a putative explanation for the l(3)ecd1 mutation.
abstract: Studies in vitro revealed that intact ring glands of Drosophila melanogaster convert tritiated cholesterol (C) and 25-hydroxycholesterol (25C) via 7-dehydrocholesterol (7dC) and 7-dehydro-25-hydroxycholesterol (7d25C), respectively, to ecdysone (E) and 2-deoxyecdysone (2dE), while both intact and homogenized ovaries synthesize only 2dE from these precursors. Emulsified 7d25C was incorporated directly into ecdysteroids by these tissue preparations at a much greater rate than was 7d25C made in situ from 25C. To probe the basis of the biochemical defect in the ecdysteroid deficient conditional mutant ecdysoneless (ecd1), the differential incorporation into ecdysteroids of C (via 7dC), and particularly of 25C (via 7d25C), was measured relative to that observed after the incubation of 7d25C directly with both wild type and mutant tissues in vitro at 30 degrees C, the restrictive temperature. Both C and 25C were equally 7,8-dehydrogenated in situ to 7dC or 7d25C, respectively, by both wild type and mutant tissues at 30 degrees C. However, the rate of subsequent conversion of either of these delta 5,7-sterol intermediates synthesized in situ to ecdysteroids was reduced an average of 50% in the mutant tissues relative to the wild type. Yet, when emulsified 7d25C was incubated directly with either the wild type or mutant tissues at the restrictive temperature, the amplified rate of conversion of the freely available 7d25C to ecdysteroid by these tissues was identical. These data suggest that the defect in ecd1 tissue-mediated ecdysteroidogenesis does not involve a "hit" on any of the enzymes involved in either the 7,8-dehydrogenation of C or 25C or in the subsequent oxidation of 7d25C or 7dC to ecdysteroid. Rather, the mutation appears to affect the expression of a gene governing the translocation of delta 5,7-sterol intermediates from the subcellular compartment where they are synthesized and/or stored to the site of subsequent oxidation to ecdysteroid.
authors.affiliation: Department of Biology, University of North Carolina, Chapel Hill 27599-3280, USA.
authors.forename: J T
authors.initials: JT
authors.lastname: Warren
authors.forename: J S
authors.initials: JS
authors.lastname: Bachmann
authors.forename: J D
authors.initials: JD
authors.lastname: Dai
authors.forename: L I
authors.initials: LI
authors.lastname: Gilbert
mesh_terms: D000818:Animals; D002784:Cholesterol / Q000031:analogs & derivatives / Q000378:metabolism*; D004331:Drosophila melanogaster / Q000378:metabolism*; D004440:Ecdysone / Q000235:genetics / Q000378:metabolism*; D026461:Ecdysteroids; D005260:Female; D007301:Insect Hormones / Q000378:metabolism*; D007553:Isotope Labeling; D007814:Larva; D010053:Ovary / Q000378:metabolism; D013256:Steroids / Q000378:metabolism*; D013261:Sterols / Q000378:metabolism; D014316:Tritium
chemical_list: D026461:Ecdysteroids; D007301:Insect Hormones; D013256:Steroids; D013261:Sterols; D014316:Tritium; D004440:Ecdysone; D002784:Cholesterol