An assay to detect in vivo Y chromosome loss in Drosophila wing disc cells.

Loss of the Y chromosome in Drosophila has no impact on cell viability and therefore allows us to assay the impact of environmental agents and genetic alterations on chromosomal loss. To detect in vivo chromosome loss in cells of the developing Drosophila wing primordia, we first engineered a Y chromosome with an attP docking site. By making use of the ΦC31 integrase system, we site-specifically integrated a genomic transgene encompassing the multiple wing hair (mwh) locus into this attP site, leading to a mwh(+)Y chromosome. This chromosome fully rescues the mwh mutant phenotype, an excellent recessive wing cell marker mutation. Loss of this mwh(+)Y chromosome in wing primordial cells then leads to manifestation of the mwh mutant phenotype in mwh-homozygous cells. The forming mwh clones permit us to quantify the effect of agents and genetic alterations by assaying frequency and size of the mwh mosaic spots. To illustrate the use of the mwh(+)Y loss system, the effects of four known mutagens (X-rays, colchicine, ethyl methanesulfonate, and formaldehyde) and two genetic conditions (loss- and gain-of-function lodestar mutant alleles) are documented. The procedure is simple, sensitive, and inexpensive.

Structural chromosomal aberrations, aneuploidy, and mosaicism in early cleavage mouse embryos derived from spermatozoa exposed to γ-rays.

PURPOSE: To quantitatively and qualitatively investigate the changes in chromosomal aberrations during early cleavage in mouse embryos derived from γ-irradiated spermatozoa. MATERIALS AND METHODS: Mature males were exposed to 2 Gy or 4 Gy of ¹³7Cs γ-rays, and their spermatozoa were used to produce embryos via in vitro fertilisation (IVF). The metaphase chromosomes were prepared from one-cell, two-cell, and four-cell embryos. In the chromosome preparations from two-cell and four-cell embryos, the separation of the sister blastomeres was precluded by treatment of the embryos with concanavalin A. The incidence of embryos with structural chromosomal aberrations, aneuploidy, or mosaicism was estimated. The fates of the different types of γ-ray-induced structural chromosomal aberrations were also investigated in those embryos. RESULTS: The exposure of spermatozoa to 2 Gy or 4 Gy γ-rays caused structural chromosomal aberrations in 25.9% and 35.7% of the resultant one-cell embryos, respectively. At two-cell embryonic stage, the incidence of structural chromosomal aberrations was 17.4% in the 2 Gy group and 27.1% in the 4 Gy group. At the four-cell embryonic stage, although the incidence of control embryos with structural chromosomal aberrations was considerably high, the net incidence of embryos with radiation-induced structural chromosomal aberrations was similar to that at the one-cell stage. The incidence of aneuploidy was high in two-cell and four-cell embryos after both doses of γ-rays. The incidence of mosaicism increased significantly in dose- and embryonic-stage-dependent manners. Anaphase lag, and the degeneration and non-disjunction of the aberrant chromosomes were frequently observed in aneuploid and mosaic embryos. CONCLUSIONS: Mouse sperm DNA is highly vulnerable to γ-rays. The structural chromosomal aberrations of sperm origin are unstable in their behaviour and structure during cleavage, and therefore cause secondary aneuploidy and mosaicism in the early cleavage embryos.

Chromatic organization of cone photoreceptors in the retina of rainbow trout: single cones irreversibly switch from UV (SWS1) to blue (SWS2) light sensitive opsin during natural development.

The retinas of salmonid fishes have single and double cones arranged in square to row formations termed mosaics. The square mosaic unit is formed by four double cones that make the sides of the square with a single (centre) cone in the middle, and a single (corner) cone at each corner of the square when present. Previous research using coho salmon-derived riboprobes on four species of anadromous Pacific salmon has shown that all single cones express a SWS1 (UV sensitive) visual pigment protein (opsin) at hatching, and that these cones switch to a SWS2 (blue light sensitive) opsin during the juvenile period. Whether this opsin switch applies to non-anadromous species, like the rainbow trout, is under debate as species-specific riboprobes have not been used to study opsin expression during development of a trout. As well, a postulated recovery of SWS1 opsin expression in the retina of adult rainbow trout, perhaps via a reverse process to that occurring in the juvenile, has not been investigated. Here, we used in situ hybridization with species-specific riboprobes and microspectrophotometry on rainbow trout retina to show that: (1) single cones in the juvenile switch opsin expression from SWS1 to SWS2, (2) this switch is not reversed in the adult, i.e. all single cones in the main retina continue to express SWS2 opsin, and (3) opsin switches do not occur in double cones: each member expresses one opsin, maximally sensitive to green (RH2) or red (LWS) light. The opsin switch in the single cones of salmonid fishes may be a general process of chromatic organization that occurs during retinal development of most vertebrates.

Genetic damage in a population of slider turtles (Trachemys scripta) inhabiting a radioactive reservoir.

Turtles inhabiting a radioactive reservoir appear to experience genetic damage due to environmental exposure to low concentrations of long-lived radionuclides. Total body burdens for the 50 reservoir turtles examined in the survey ranged from 164.7-4679.3 Bq for cesium-137 and from 462.6-5098.3 Bq for strontium-90. Flow cytometric (FCM) assays of red blood cell nuclei demonstrated significantly greater variation in DNA content for the reservoir turtles than for turtles from a nearby, non-radioactive site. Furthermore, two of the reservoir turtles possessed FCM profiles that are indicative of aneuploid mosaicism. These data strongly suggest that exposure to low-level radiation may involve a sensitive genetic response in a natural population.

[Specificity of replicating instability in Schizosaccharomyces pombe haploid yeasts]. / O spetsifichnosti replitsiruiushcheisia nestabil'nosti u drozhzhei Schizosaccharomyces pombe.

UV-induced genetic instability in haploid Schizosaccharomyces pombe does not appear to be very locus-specific. This conclusion contradicts the data previously published by other authors. The possible causes for this discrepancy are discussed.

Testing the mutagenicity of malondialdehyde and formaldehyde by the Drosophila mosaic and the sex-linked recessive lethal tests.

The mutagenicities of malondialdehyde and formaldehyde were tested by screening each for genetic mosaics of Drosophila melanogaster and by the Muller-5 test for sex-linked recessive lethal mutations. For comparison, the effects of X-rays were also assayed by the above technique. Malondialdehyde, a degradation product of polyunsaturated fatty acids, was found to be a weak mutagen by the above criteria; it induced point mutations and chromosome exchanges at low frequency, as proved by the mosaic test, but failed to induce detectable sex-linked lethality. Formaldehyde was more mutagenic than malondialdehyde; beside induction of mosaic spots it induced sex-linked recessive lethal mutations, but only in the larval testes of Drosophila. Formaldehyde also induced disintegration of the clones. Formaldehyde treatment (feeding larvae with formaldehyde-containing food for about 4 days) was 5 times more mutagenic than malondialdehyde treatment and 5 times less effective than irradiation by 1000 R of X-rays. Wing mosaicism offers a more sensitive way to detect mutagenesis as compared with eye mosaicism. It is suggested that aldehyde-induced mosaic spots derive from mitotic recombination and point mutations.

[Analysis of somatic mosaicism in the Drosophila melanogaster radiosensitive mutant rad(2)201G1]. / Analiz somaticheskogo mozaitsizma u radiochuvstvitel'nogo mutanta rad(2)201G1 Drosophila melanogaster.

Using heterozygosity for bw mutations, the frequency of somatic mosaicism has been studied in Drosophila melanogaster flies, homozygous for radiosensitive mutation, rad(2)201G1, and in those not carrying the mutation. Data obtained for control and after irradiation of larvae with 150, 450 and 750 R showed that homozygotes for the rad(2)201G1 were characterized by elevated levels of both spontaneous and induced mosaicism.

Somatic mosaicism in plants with special reference to somatic crossing over.

Plant systems in use for the detection of environmental mutagens appear capable of detecting all types of genetic effects which can be studied in animals. The study of somatic mosaicism, however, is better developed in plants than in higher animals. A case is presented here which shows the ability of plant systems in analyzing a host of genetic end points, including chromosome aberrations like deletions, somatic crossing over, numerical inequality, gene conversion, paramutations and point mutations. The systems in general use utilize certain varieties of Tradescantia, Glycine max, Nicotiana tabacum, Antirrhinum majus, Petunia hybrida, and Arabidopsis thaliana. Heterozygous plants or their homozygous counterparts with gene markers affecting chlorophyll development or anthocyanin in floral parts are exploited in these studies. Mutagens produce different frequencies of different types of spots typical of the mode of action of the agent. Analysis of these parameters may be used to predict, at least qualitatively, the kind of genetic damage that might be produced in man. Besides, one can test the validity of interpretation by traditional progeny tests of plants raised from tissue culture from sectors as in Nicotiana and/or by precursor analysis as done in Antirrhinum. The study of mosaicism in plants offers quite inexpensive, rapid, and reliable tests of mutagenicity at least as a preliminary eukaryotic test system.

Effects of treating drosophila females with TEM on their mortality, and the yields of mutation recovered from x-irradiated sperm.

Ring-X-bearing Drosophila males were irradiated with 0, 1000, 2000 or 3000 R of X-rays and mated to females that had been injected with saline or 10(-4) M TEM. The mortality and the fertility of the treated females were recorded. The rate of dominant lethals, of entire sex chromosome loss and partial loss of the Y chromosome, the sex ratio, and the rate of mosaics were determined on the progency. TEM slightly increased the rate of mortality of the females. But it did not influence the yield of mutations recovered from the irradiated spermatozoa.