Bertil Rasmuson

Insect immunity: Inducible antibacterial activity in Drosophila

Adult Drosophila melanogaster were found to produce an inducible antibacterial activity as a response to an injection (vaccination) of the non-pathogenic bacterium, Enterobacter cloacae. Vaccinated flies showed an increased survival time after a second injection with an insect-pathogenic bacterium, Pseudomonas aeruginosa. This immune response was blocked by cycloheximide, an inhibitor of eukaryotic protein synthesis, a fact which indicates de novo synthesis of the antibacterial factor operating in vivo. Electrophoretic mobility at pH 4 in combination with antibacterial assay and immunological cross reactivity was used to demonstrate attacin-like factors in hemolymph and cecropin-like factors in cell-free extracts of whole flies. Extract of flies also contained lysozyme but this enzyme could not be induced. Some active material is pre-existing and can be released by treatment of whole flies with ultrasound (sonication) or microwaves. Compared to wild-type flies, a mutant strain deficient in leucine aminopeptidase showed much higher values for antibacterial activity and lysozyme.

Genetic instability in Drosophila melanogaster

SummaryAn unstable long tandem duplication which includes the white locus twice, marked with wsp in the left and w17G in the right locus, when kept in males has been found to produce red-eyed sons which have lost the long duplication and with it the wsp and w17G mutants. Such exceptions were produced also when w17G had been exchanged for wa.Stocks originating from these exceptions are unstable, producing: 1) zeste males, also unstable, 2) w- deletions, stable, 3) transpositions of the white locus to sites in other chromosomes.The instability is interpreted as the effect of an IS element, within or adjacent to the white locus, which is supposed to retain a duplication of the proximal zeste interacting part of this locus. According to the orientation of the IS element the duplicated part can be active or inactive, giving a zeste or red eye phenotype.The frequency of exceptional offspring after X-ray treatment of the red and zeste unstable stocks have been compared to stable stocks with corresponding genotypes.

Mutagenic effects of petrol in Drosophila melanogaster I. Effects of benzene and 1,2-dichloroethane

Commercial petrol and two of its components, benzene and 1,2-dichloroethane, were tested for mutagenicity in Drosophila melanogaster. The chemicals were given to larvae through their food supply. A genetically unstable sexlinked test system caused by a transposable genetic element was used. Mutagenicity was measured by the frequency of somatic mutations for eye pigmentation. Commercial petrol and 1,2-dichloroethane showed mutagenic activity. With the system used, benzene did not show any mutagenic activity. The high frequency of mutations induced by 1,2-dichloroethane indicate the existence in Drosophila of a metabolic activating system.

The use of a mutationally unstable X-chromosome in Drosophila melanogaster for mutagenicity testing.

Somatic eye-colour mutations in an unstable genetic system, caused by a transposable element in the white locus of the X-chromosome in Drosophila melanogaster, is suggested as an assay system for mutagenicity testing. The system is evaluated by comparison with a corresponding system in a stable X-chromosome. Its sensitivity is confirmed with X-ray and EMS treatment, and it is found to be confined to the specific segment of the X-chromosome where the transposable element is localized.

A negative test for mutagenic action of microwave radiation in Drosophila melanogaster.

Microwave radiation (2450 MHz CW) was tested for mutagenicity in Drosophila melanogaster. Embryos in water were exposed to the electromagnetic field with a mean specific absorption rate of 100 W/kg. A sensitive somatic test system was used, in which mutagenicity was measured as the frequency of somatic mutations for eye pigmentation. With the test system used, microwaves did not show any mutagenic activity.

Uptake and incorporation in pteridines of externally supplied GTP in normal and pigment-deficient eyes ofDrosophila melanogaster

Some aspects of the synthesis of drosopterins in the eyes ofDrosophila melanogaster have been studied in flies with different levels ofwhite gene expression. The activity of GTP cyclohydrolase was found to differ between wild-type and yellow-eyed mutantsin vivo but notin vitro. To elucidate the uptake of substrate, we measured the removal of labeled GTP from the incubation medium by excised pupal eyes and followed the subsequent fate of this label. It was found that GTP was dephosphorylated to guanosine extracellularly before label was taken up by the eye tissue. The uptake was much lower in yellow and white eyes than in wild-type eyes, and in the latter, a considerable part of the label was present in pteridine compounds. The strain differences in the uptake of label seem to be due to different rates of intracellular utilization of guanine derivatives in pteridine synthesis. We suggest that this is caused by a hampered transport of precursor (possibly GTP) in white andzeste eyes through the membrane of red pigment granules.

Enhanced susceptibility of a transposable-element-bearing strain of Drosophila melanogaster to somatic eye-color mutations by ethyl nitrosourea, methyl nitrosourea, and X-rays.

A strain of Drosophila with the genes z and w+ plus a transposable element (TE) is about 3 times more sensitive than a strain without TE toward somatic eye-color mutations after larval exposure to ethyl nitrosourea, methyl nitrosourea and X-rays. The assay system with TE is simple, reliable, and sensitive for detecting somatic mutations induced in vivo by mutagens.