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Farnesylacetone is a natural terpene extracted from androgenic glands of the crustacean Carcinus maenas and is capable of inhibiting proliferation, notably in transformed mammalian cells. Flow cytometry with three lipophilic probes, diphenylhexatriene, trimethylammonium-diphenylhexatriene, and Nile red, has revealed modifications of the lipidic metabolism in transformed FR3T3-mTT4 rat fibroblasts treated by farnesylacetone, including changes in membrane fluidity. Farnesylacetone strongly increased the number of neutral lipidic droplets in the cytoplasm. Moreover, after prolonged terpene treatment, the membrane fraction of cells contained a substantial level of triglycerides. Farnesylacetone provoked an immediate but transitory increase in membrane fluidity of the cell membrane. The change in measured lipid fluidity appears to be due to these triglycerides rather than to the phospholipids.

The Y organ of Carcinus mediterraneus parasitized by Sacculina carcini and of normal Crabs at the C4 stage of the intermolt cycle have the same ultrastructure. At this stage the secretory activity of the Y organ appears to be reduced.

Subcultures of ovaries and testis of the crab Carcinus maenas have been performed in the presence of L-[Me-14C]methionine. Introduction in the medium of a chromatographically-purified liposoluble fraction from the androgenic glands of the same animal inhibits the biological methylation of the tRNA of the ovaries by 62%. The inhibition of methylation of five individual bases varies from 45% to 84%. No inhibition of tRNA methylation is observed under the same conditions with testis subcultures.

t, t-farnesylacetone 1 and hexahydrofarnesylacetone 2 have been previously identified in extracts from the androgenic gland of the male Crab Carcinus maenas. These compounds inhibit in vitro the methylation of E. coli B tRNA and of Calf thymus histones with S-adenosylmethionine methyl-14C as methyl donor and methylases from Crab testis. Rat liver or a 1-adenine methylase from a Mouse plasmocytoma (1 is approximately 200 times more active than 2).

Radioimmunoassay is performed for a quantitative study of ecdysone in Carcinus moenas. Two periods can be determined in the moulting cycle. In haemolymph from ecdysis until beginning D0 stage, the moulting hormone content is not very high; from D0 stage until D4 stage, the hormone titer rises considerably. We can observe a similar rise in hormone content in both haemolymph and Y organ during the premolt stages. In sacculinized Crabs, the determined values of ecdysone in haemolymph and in Y organ are lower than those determined in healthy animals at the same stage.

Regeneration was induced in the imaginal discs in situ following lesions caused by heat-sensitive cell-lethal mutations. A clonal analysis of this event demonstrated that the subsequent delay in pupariation was correlated with the amount of extra growth that occurred during the regeneration. Pupariation of heat-treated gynandromorphs bearing the mutations was also retarded, and the duration of larval development increased with greater amounts of mutant tissue, it was therefore correlated with the extent of the lesions in the imaginal discs. Elimination of entire imaginal discs, or the presence of very small amounts of lethal tissue, did not result in prolonged larval life.

The temperature-sensitive 1(3)ecd-1ts mutation (A. Garen, L. Kauvar, and J.A. Lepesant (1977). Proc. Natl. Acad. Sci USA 74, 5099-5103.) has been used in several laboratories to obtain Drosophila larvae deprived of moulting hormone. The development of mutants and controls during the third larval instar at permissive (20 degrees C) and restrictive temperatures (29 degrees C) was compared. Pupariation was inhibited when larvae were shifted to the restrictive temperature immediately at the second moult. The permanent larvae obtained remained active, did not leave the food, and reached a maximum weight superior to the weight of controls. Ecdysteroids were studied during the third larval instar by HPLC analysis and radioimmunoassays. A careful synchronization of the larvae at the second moult enabled the confirmation that at least one ecdysteroid peak occurs during the third larval instar, prior to the wandering stage in controls (20 or 29 degrees C). Ecdysone was then the predominant moulting hormone, whereas 20-hydroxyecdysone was the main ecdysteroid at the time of pupariation. Low levels of ecdysteroid were measured in mutant larvae shifted to 29 degrees C immediately at the second moult but larvae completely deprived of immunoreactive material were never observed. Nearly normal levels of ecdysteroids appeared at 27.5 degrees C. Feeding ecd-1 larvae maintained at restrictive temperature on 20-hydroxyecdysone-yeast mixture for 16 hr triggered abortive pupariation. Ecdysteroid levels were measured after the return of the larvae to the standard medium; normal levels were restored 24 hr later. The mutant ecd-1 appears to present interesting opportunities for the detailed study of the hormonal induction of a developmental process during the third larval instar.(ABSTRACT TRUNCATED AT 250 WORDS)

The proteic composition of haemolymph from C. mediterraneus varies during the intermoult cycle. Some proteic fractions are occasionally absent in post-ecdysis. At each intermoult stage, parasitized Crabs have generally the same protein pattern as healthy ones, but the presence of the observed fractions is not constant. Compared healthy Crabs, the sacculinized animals show a surnumerary proteic fraction. With very short delay, the injection of ecdysterone puts the proteic composition of haemolymph from sacculinized Crabs in order, this composition becomes comparable to that from healthy individuals.

Extracts from muscles, testis, seminal vesicles and ovaries of the Crab, Carcinus maenas, have been studied in vitro, in presence of [14C]-methyl S-adenosylmethionine, with an E. coli tRNA as methyl acceptor. The highest level of methylases is found in the testis. It has been reported previously that a purified fraction extracted from the androgenic glands of Carcinus maenas inhibits the vitellogenesis in ovaries. We now show that the same fraction inhibits tRNA methylation in an extract of testis as methylase; a 50% inhibition is obtained with about 10 mug of a purified fraction corresponding to 15 glands. With an enzymatic preparation from the ovaries, a 50% inhibition of the tRNA methylase is observed with the purified extract from 4 glands.

Farnesylacetone (C18 H30 0) is a male hormone extracted from the androgenic gland of crab, Carcinus maenas. Appropriate enzymatic assays, as well as spectrophotometric studies, indicate that micromolar concentrations of farnesylacetone interact with the electron transport pathway of rat liver mitochondria. By the use of artificial electron donors and electron acceptors, it is shown that farnesylacetone immediately inhibits the electron transfer within complex I (NADH ubiquinone reductase activity) and complex II (succinate ubiquinone reductase activity). It is proposed that farneylacetone could interact with these two complexes of the respiratory chain at the level of the iron-sulfur centers implicated in the dehydrogenase activities. These observations are compared with the results obtained with terpenic molecules which interact with mitochondrial respiration.