Incorporation of [14C]5-aminolevulinate and [3H]leucine into cytochrome c, biliprotein and total soluble protein was followed from the last larval instar to the adult stage in Pieris brassicae. The titer of ecdysteroids during the pupal stage was determined with a radioimmunoassay to correlate synthesis of heme products and of protein(s) with adult differentiation. Incorporation of both precursors showed a developmental profile with high synthetic activities in feeding larvae and in pupae after the release of ecdysteroids. Variation of the hormone titer during pupal life differed significantly in males and females. Labeling of cytochrome c by both 14C and 3H was as expected from the variation of its concentration reported in a preceding paper; highest in corporation was around adult emergence. The results demonstrate that i) the accumulation of cytochrome c in the developing adult insect is primarily due to de novo synthesis of both heme and apocytochrome c, performed under coordinate control, and ii) the concentration of 5-aminolevulinate is not rate-limiting in the formation of cytochrome c. Biliverdin IXγ, the major tetrapyrrolic product in this insect, seems to be directly derived from (free) heme and relatively short-lived as deduced from a time-course study. Formation of the bilin, i.e. destruction of heme, increased concomitantly to the initiation of adult differentiation by ecdysteroids in the pupa but later decreased at adult emergence. Synthesis of cytochrome c takes place as a late event during terminal development. Thus, the pathways leading to the two major heme products seem to be differently regulated during development.

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