The relationship between radiation-induced and transposon-induced genetic damage during Drosophila oogenesis

The combined effect of X-irradiation and transposon mobility on the frequencies of X-linked recessive lethals and dominant lethals was investigated in female hybrids in the P-M system of hybrid dysgenesis. X-linked lethals were measured in G2 hybrid dysgenic females whose X chromosome was derived from the M X P cross. To test for additivity or synergism, the mutation rate in irradiated dysgenic females was compared to that of unirradiated females as well as to irradiated nondysgenic hybrid females derived from M X M crosses. Eggs collected for 2 days after irradiation, were represented by the more radiation-sensitive A and B oocytes (about 75%) and the least sensitive C oocytes (about 25%). The production of X-linked lethal events in X-irradiated dysgenic females was 8.1%, as compared to 4.5% in dysgenic controls and 3.4% in irradiated, nondysgenic controls, demonstrating an additive effect of radiation and dysgenesis-induced genetic damage. The effect of irradiation on sterility of dysgenic hybrid females was a negative one, resulting in 20% less sterility than expected from an additive effect. The combined effect of radiation and dysgenesis on dominant lethality tested in A, B and C oocytes of the same hybrid females was synergistic. Egg broods collected for 3.5 days after irradiation showed that significantly more damage was induced in the presence of ionizing radiation in dysgenic females than in their nondysgenic counterparts. This effect was most obvious in B and C oocytes. The synergism observed may be related to the inability of cells to repair the increased number of chromosome breaks induced both by radiation and transposon mobility.     

The relationship between radiation-induced and transposon-induced genetic damage during Drosophila spermatogenesis

The combined effect of transposon mobility and X-rays on X-linked recessive lethals and dominant lethals was measured in the germ line of F₁ male hybrids in the P-M system of hybrid dysgenesis. X-linked lethal mutation rate was measured in the chromosome derived from the P-strain father of the M × P cross. Mutations induced in irradiated dysgenic males were compared to those of unirradiated males, as well as to irradiated nondysgenic males derived from M × M crosses. Three four-day broods of sperm were tested for both X-linked lethals and dominant lethals. X-linked lethal mutation rate in dysgenic control males was 6.38%, 6.36% and 4.55% in broods 1, 2 and 3 respectively, thus showing a decrease in older males. The mutation rate in the same broods of irradiated, nondysgenic control males was 3.66%, 4.46% and 6.38%, respectively. The rate obtained in dysgenic irradiated males was 10.33, 11.16 and 7.97 in the same 3 broods. These results demonstrate that when X-rays and P element mobility were and 7.97 in the same 3 broods. These results demonstrate that when X-rays and P element mobility were combined as a source of mutagenesis, a strickly additive effect on genetic damage was observed in the first two broods of sperm which represent primarily mature sperm and spermatids respectively. The third brood, representing mostly spermatocytes showed a less than additive effect, probably due to germinal selection. In contrast, the induction of dominant lethals showed a clearly synergistic effect in the last two Broods of sperm tested, when X-rays and transposon mobility were combined. The X-ray component of dominant lethlity in brood 1, representing mostly mature spermatozoa, was negative, indicating a lower than expected lethality induced by X-irradiation in the presence of P element mobility. The X-ray-induced component of dominant lethality, was expressed as the per cent of embryo lethality after adjusting the results obtained with each brood of sperm from nondysgenic and dysgenic males to their respective unirradiated controls. These values were 32.3%, 30.5% and 64.7% for brood 1, 2 and 3 respectively from nondysgenic males, and 14.1%, 56.1% and 71.4% for the same broods from dysgenic males. Thus the differential effect of X-rays in sperm broods 1, 2 and 3 was −18.2, +25.6 and +6.7% respectively. These results suggest that the synergistic effect may be due to the common component of X-ray and P element-induced genetic damage, namely chromosome breaks, and that the interaction of these lesions resulted in a greater than additive number of of unrestitude chromosome breaks and nonviable chromosomal rearrangements.     

Radiation and transposon-induced genetic damage in Drosophila melanogaster: X-ray dose-response and synergism with DNA-repair deficiency.

The interaction of X-ray-induced and transposon-induced damage was investigated in P-M hybrid dysgenesis in Drosophila melanogaster. The X-ray dose-response of 330-1320 rad was monitored for sterility, fecundity and partial X/Y chromosome loss among F2 progeny derived from the dysgenic cross of M strain females xP strain males (cross A) and its reciprocal (cross B), using a weaker and the standard Harwich P strain subline. The synergistic effect of P element activity and X-rays on sterility was observed only in cross A hybrids and the dose-response was nonlinear in hybrids derived from the strong standard reference Harwich subline, Hw. This finding suggests that the lesions induced by both mutator systems which produce the synergistic effect are two-break events. The effect of increasing dose on the decline of fecundity was synergistic, but linear, in hybrids of either subline. There was no interaction evident and thus no synergism in X/Y nondisjunction and in partial Y chromosome loss measured by the loss of the Bs marker alone or together with the y+ marker. Interaction was detected in the loss of the y+ marker alone from the X and Y chromosomes. The possible three-way interaction of X-rays (660 rad), post-replication repair deficiency and P element mobility was assessed by measuring transmission distortion in dysgenic males derived from the II2 P strain. X-Irradiation of spermatids significantly increased the preferential elimination of the P-element-bearing second chromosome in mei-41, DNA-repair-deficient dysgenic males, but had no effect in their DNA-repair-proficient brothers. These findings indicate that the post-replication repair pathway is required for processing lesions induced by the combined effect of P element mobility and X-rays, and that the unrepaired lesions ultimately lead to chromosome loss.     

Radiation-induced translocations in mice: persistence, chromosome specificity, and influence of genetic background

The translocation frequency response in the chromosomes of peripheral blood lymphocytes is widely used for radiation biomonitoring and dose estimation. However, this assay is based upon several assumptions that have not been rigorously tested. It is typically assumed that the translocation frequency in blood lymphocytes reflects the level of genomic damage in other hemopoietic tissues and is independent of the chromosome probe and genetic background. We conducted studies to evaluate these assumptions using mice with different genetic backgrounds. Six different whole-chromosome fluorescence in situ hybridization (FISH) probes were used to detect translocations in peripheral blood lymphocytes at multiple times after whole-body irradiation. Translocation frequencies were chromosome-independent at 6 and 16 weeks after exposure but were chromosome-dependent at 1. 5 years after exposure. Similar translocation frequencies were observed in blood, bone marrow and spleen at 1.5 years, supporting previous suggestions that genetically aberrant peripheral blood lymphocytes may derive from precursor populations in hemopoietic tissues. Translocations measured 66 h after irradiation differed among some strains. We conclude that the translocation frequency response is a complex phenotype that is influenced not only by exposure dose but also by genetic background, the choice of chromosome analyzed, and time after exposure. These results raise important considerations for the use of the FISH-based translocation frequency response for radiation dosimetry and biomonitoring.     

Sex-of-offspring-specific transmission ratio distortion on mouse chromosome X

During our study of the DDK syndrome, we observed sex ratio distortion in favor of males among the offspring of F(1) backcrosses between the C57BL/6 and DDK strains. We also observed significant and reproducible transmission ratio distortion in favor of the inheritance of DDK alleles at loci on chromosome X among female offspring but not among male offspring in (C57BL/6 x DDK)F(1) x C57BL/6 and (C57BL/6-Pgk1(a) x DDK)F(1) x C57BL/6 backcrosses. The observed transmission ratio distortion is maximum at DXMit210 in the central region of chromosome X and decreases progressively at proximal and distal loci, in a manner consistent with the predictions of a single distorted locus model. DXMit210 is closely linked to two distortion-controlling loci (Dcsx1 and Dcsx2) described previously in interspecific backcrosses. Our analysis suggests that the female-offspring-specific transmission ratio distortion we observe is likely to be the result of the death of embryos of particular genotypic combinations. In addition, we confirm the previous suggestion that the transmission ratio distortion observed on chromosome X in interspecific backcrosses is also the result of loss of embryos.     

Radiation-induced cytogenetic damage in relation to changes in interphase chromosome conformation

The premature chromosome condensation (PCC) technique was used to study several factors that determine the yield of chromosome fragments as observed in interphase cells after irradiation. In addition to absorbed dose and the extent of chromosome condensation at the time of irradiation, changes in chromosome conformation as cells progressed through the cell cycle after irradiation affected dramatically the yield of chromosome fragments observed. As a test of the effect of chromosome decondensation, irradiated metaphase Chinese hamster ovary (CHO) cells were allowed to divide, and the prematurely condensed chromosomes in the daughter cells were analyzed in their G1 phase. The yield of chromosome fragments increased as the daughter cells progressed toward S phase and chromosome decondensation occurred. When early G1 CHO cells were irradiated and analyzed at later times in G1 phase, an increase in chromosome fragmentation again followed the gradual increase in chromosome decondensation. As a test of the effect of chromosome condensation, G0 human lymphocytes were irradiated and analyzed at various times after fusion with mitotic CHO cells, i.e., as condensation proceeded. The yield of fragments observed was directly related to the amount of chromosome condensation allowed to take place after irradiation and inversely related to the extent of chromosome condensation at the time of irradiation. It can be concluded that changes in chromosome conformation interfered with rejoining processes. In contrast, resting chromosomes (as in G0 lymphocytes irradiated before fusion) showed efficient rejoining. These results support the hypothesis that cytogenetic lesions become observable chromosome breaks when chromosome condensation or decondensation occurs during the cell cycle.     

Radiation-induced inversions and reciprocal translocations in Anopheles atroparvus

Inversions and reciprocal translocations were induced in Anopheles atroparvus by irradiation of males with X-rays. 22 aberrations were produced in stocks and were identified as follows: 6 paracentric, 6 pericentric inversions and 10 reciprocal translocations (9 autosomal and 1 sex-linked). Partial sterility in the offspring of this stock is demonstrated. The practical significance of constructing stocks with inversions and translocations for genetic control of pest insects is considered.     

Recombination in the X chromosome of Drosophila melanogaster females heterozygous for two autosomal translocations

X chromosome recombination was measured in females carrying two 2; 3-translocations. Total X chromosome recombination values varied according to the amount of structural heterozygosity between the two translocations. The results support the hypothesis that the observed effects of autosomal translocation homozygosity on recombination in the X chromosome are due to homozygosity for position effects of the translocation breakpoints and are not due to chromosome discontinuity.     

Radiation-induced nondisjunction and Robertsonian translocation in Drosophila

In Drosophila melanogaster, gametes formed by oocytes in which Robertsonian translocations were induced in an immature stage usually show chromosomal imbalance. It is estimated that fewer than 20% of the gametes bearing newly induced Robertsonian translocations “fusing” X and fourth chromosomes are of balanced constitution. In contrast, when the two acrocentric pairs, X and fourth chromosomes, are replaced by an X-4 Robertsonian translocation, treatment of immature oocytes of homozygotes produces some 5–6-fold fewer sex-chromosome trisomics than do females of normal karyotype. In the place of such trisomics (having separate sex chromosomes), there is a much smaller number of compound-X chromosomes formed and a number of compound-fourth chromosomes as well. However, the production of “XO” males is not appreciably smaller in the translocation homozygotes. A number of possible mechanisms to account for this are suggested. The findings are consistent with the expectations of the hypothesis that radiation-induced nondisjunction results from improper conjunctions of heterologues, brought about by chromatid interchange^{7–12, 16}.     

Paternal transmission of entire compounds of chromosome two in Drosophila melanogaster

Summary In Drosophila melanogaster, entire compound second chromosomes (2R2L·2L2R) consist of the entire amount of genetic material normally found on separate homologues, as well as significant amounts of heterochromatin derived from the Y chromosome, joined to a single centromere. Genetic analysis demonstrates that information carried upon the Y chromosome influences the rate of transmission of the compound in the male.This work was supported in parts by grants E-501536 and GM-18678 from the National Institutes of Health     

P element transposon-induced quantitative genetic variation for inebriation time in Drosophila melanogaster

Summary Bi-directional selection was carried out in coisogenic stocks with and without mobilised P element transposons to determine whether P elements induce quantitative genetic variation for inebriation time in Drosophila. There was significant response to 11 generations of selection in both pairs of replicates of bi-directional selection from an isogenic base stock in which P elements had been mobilised. Conversely, there was no significant response to 11 generations of identical selection in the control lines derived from a relatively inbred line lacking P elements. Thus, P elements have induced quantitative genetic variation for inebriation time.     

Radiation-induced translocations in spermatogonial stem cells of Macaca fascicularis and Macaca mulatta

3 adult monkeys, one Macaca fascicularis and two Macaca mulatta, were whole-body irradiated with 1 Gy gamma-rays (60 Co). Reciprocal translocations induced in spermatogonial stem cells were scored as translocation multivalents in primary spermatocytes from 7.5 to 27.5 months after exposure. The translocation yields ranged from 4.1% at the earliest to 1.8% at the latest sampling interval. No significant differences were observed in the responses of the individual animals. A decline in the translocation frequencies with time after treatment was found in all 3 animals. The present data are different from those reported for testicular X-irradiation of the rhesus monkey Macaca mulatta (van Buul, 1980; Lyon et al., 1976) in that the translocation yields are higher. They are consistent with the results reported for testicular gamma-irradiation of the crab-eating monkey Macaca fascicularis (Matsuda et al., 1984, 1985). In view of the present results it appears unlikely that a species difference exists within the genus Macaca in the sensitivity of spermatogonial stem cells to the induction of translocations by ionizing radiation.     

The distribution of chromosome damage, non-reciprocal translocations and clonal aberrations in lymphocytes from Chernobyl clean-up workers

In this paper we determined whether the frequencies of translocations and insertions are proportional to chromosome size in peripheral blood lymphocytes from Chernobyl nuclear accident clean-up workers and healthy unexposed control subjects. The frequency of aberrations among chromosomes 1, 2 and 4 in both groups was found to be significantly different from the distribution expected on the basis of chromosome size, although the difference was only marginally significant in controls. We also determined whether differences exist in aberration frequencies measured by two scoring systems: the classical method, where reciprocal exchanges are scored as one event, and PAINT, where each break junction is scored as a single event. The two scoring systems gave highly correlated results which yielded an interpretable arithmetic relationship between frequency measurements using the two systems. Approximately 34% of all translocations were observed to be non-reciprocal, and cells bearing clones of abnormal cells were observed in 6 of 198 subjects (3.0%). Our results demonstrate that clones of abnormal cells and the presence of non-reciprocal translocations contribute to the non-proportional distribution of radiation-induced and spontaneous cytogenetic damage.