Abstract

The effects of cycloheximide, chloramphenicol and ethidium bromide on the blowfly Calliphora erythrocephala were studied. In the first set of experiments, toxic levels were determined by examining activity and mortality of flies after injection of various doses of each drug. In the second set of experiments, the effect of drug treatment on flight muscle mitochondrial protein synthesis was determined in relation to age by following the incorporation of radioactively labelled amino acid, [3H]leucine, into mitochondrial protein in vivo. To confirm the developmental changes in flight muscle mitochondria, mitochondrial protein content per fly was estimated from emergence to 30 days of age; the highest protein level was recorded between 6 and 10 days of age. Maximum incorporation of labelled amino acid was found in newly emerged flies, and this age group was also the most sensitive to drug treatment. By the time flies had reached 6--10 days of age, amino acid incorporation had declined to about two-thirds of the rate obtained with newly emerged flies. With 6--10-day old flies, however, the highest value for flight muscle mitochondrial protein per fly was recorded, and these flies also displayed the greatest resistance to drug treatment of any age group investigated. For example, inhibition of protein synthesis following injection of 300 micrograms/fly of chloramphenicol was only about 15% below the untreated control in 6-10-day-old flies, whereas in all other age groups investigated, inhibition ranged between 30 and 50% of the controls. At 15--20 days of age, protein synthesis decreased to a third of the newly emerged flies' rate and continued to decrease further in the 30--35-day-old group, where it was less than one sixth of the youngest age group. The effect of drug treatment on these older flies was also less than that observed with newly emerged flies, especially after chloramphenicol and ethidium bromide injections. The effect of cycloheximide however, was much the same in all age groups, with inhibition of protein synthesis being 80--90% of controls. Surprisingly, cycloheximide (1--10 micrograms/fly) had little initial effect on mortality of young flies, despite almost complete blockage in the synthesis of mitochondrial proteins at these concentrations. 95% mortality occurred only when doses of 20 micrograms/fly were given. In contrast, high doses of chloramphenicol (400 micrograms/fly) and ethidium bromide (15 micrograms/fly) caused almost total mortality a few hours after injection, although such doses never induced more than about 50% inhibition of mitochondrial protein synthesis. Each drug therefore has a different site of inhibition and induces different mortality effects. Possible explanations for these differences in mortality are discussed.

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