Studies on non-disjunction of the major autosomes in Drosophila melanogaster. I. Methodology and rate of induction by x-rays for the compound second chromosome

The induction of non-disjunction by X-irradiation of the second chromosome in stage-7 oocytes of Drosophila melanogaster has been studied by employing isochromosome stocks. This makes the quantitative recovery possible of progeny resulting from disomic and nullosomic eggs. Determination of egg hatchability has been used to correct for varying degrees of segregation in males carrying different isochromosomes. Even at exposures as low as 250 R the frequency of non-disjunction is significantly higher than in the controls. No evidence has been obtained for the existence of a threshold. In the stage-7 oocytes, the induction of non-disjunction increased linearly with radiation exposure over a range of 250–3000 R and thus seems to reflect a single-hit event. These findings could be of significance for the evaluation of genetic radiation hazards in man. In slightly younger oocyte stages the induction of disomic eggs followed dose-square kinetics. The frequency of nullosomic eggs rises exponentially with radiation exposure, presumably as a consequence of increasing chromosome loss resulting from unrestituted breaks in each of the two maternal isochromosomes. Furthermore, it was observed that the late stage-7 oocytes were more sensitivie to the induction of non-disjunction than earlier stages.     

Time dependence of Elkind-type recovery in class B oocytes of Drosophila melanogaster.

Recovery from X-ray-induced damage in class B oocytes of Drosophila melanogaster was studied by the dose-fractionation technique. A total dose of 500 R was delivered either as a single exposure or as two fractions of 2000 R and 3000 R separated by increasing time intervals. The use of attached-X females made it possible to study simultaneously the induction of dominant lethals and of chromosome aberrations (detachments of the attached-X chromosome). The same repair kinetics were observed for sublethal damage and for the lesions leading to detachments. The time-response curves are of similar shape: a plateau is reached within 20 to 30 min and half of the repairable damage disappears in 5 to 7 min. It is concluded that the same type of X-ray-induced primary lesion in chromosomes is responsible for the induction of detachments and for dominant lethals. As primary lesions actual chromosome breaks or lesions leading to breaks and chromosome rearrangements are assumed.     

Propranolol, atenolol, and trifluoperazine reduce the spontaneous occurrence of meiotic diploid products in Saccharomyces cerevisiae.

The effect of atenolol, propranolol, trifluoperazine, and caffeine on the occurrence of meiotic diploid and disomic products in Saccharomyces cerevisiae was investigated. We demonstrated that atenolol, propranolol, and trifluoperazine reduce the occurrence of meiotic diploid products and that propranolol also slightly decreases the spontaneous frequency of disomics. On the other hand, caffeine appears to be a powerful inducer of diploid meiotic products, but also shows a lesser effect on disomic induction. Since spontaneous or caffeine-induced diploids arise from a failure of the second meiotic division, it appears that the target of these drugs is at the beginning of the second meiotic division. The only common effect of trifluoperazine and propranolol, mainly investigated in mammals, was an inhibition of calmodulin activity via direct interaction. We tend, therefore, to believe that calmodulin activity must be a crucial point for the second meiotic division to begin. The increased induction of diploids, due to caffeine, may be interpreted as a consequence of an increased cyclic AMP level.     

X-ray-induced translocations in spermatoginia II. Fractionation responses in mice

The dose-response curve for reciprocal translocations induced by X-rays in spermatognial stem cells, and observed in primary spermatocytes of mice, is “hymp-shaped”, with a maximum yield at about 600 R. To test the hypothesis that the decrease in yield with increasing dose above 600 R is a consequence of the different sensitivities of cells in different stages of the cell cycle to both cell killing and chromosome aberration induction, several fractionation experiments were carried out.A total dose of 2800 R was given in repeated doses of 400 R, separated by 8-week intervals. The yield of translocations is that expected for additivity; for example, the yield at 1600 R is approximately equal to that for four separate 400-R doses.When the total dose (500 R) which gives a translocation yield on the ascending part of the dose-response curve is given as two equal fractions separated by intervals of 30, 90, or 150 min, the translocation yield decreases with increasing interval. However, when a total dose (1000 R) which would give a translocation yield on the descending part of the dose-response curve is given in two equal fractions separated by intervals of from 30 min to 6 weeks, the response is different; the translocation yield increases with intervals up to 18 h, then decreases with intervals up to 4 weeks, and finally increases again to a yield equal to additivity with an interval of 6 weeks. These changes in translocation yield with changes in interval between the two doses are explained in terms of the differential sensitivity of cells to killing and aberration induction in the different phases of the cell cycle, and by assuming that the cells surviving the first dose and repopulating the testis different cycle characteristics from normal cells.     

Translocations in mouse spermatogonia after exposure to unequally fractionated doses of x-rays

The influence of various X-ray fractionation regimes on translocation induction in mouse spermatogonia was studied. Splitting a single exposure of 600 R into two fractions of 300 R separated by 24 h did not influence the translocation yield (7.58% vs. 8.35%). When the dose was given in the unequal fractions,100 R+500 R and 500 R+100 R, with the same 24 h between the fractions, the translocation frequency was significantly altered (10,19 and 4.94%, respectively). As a possible explanation of these findings it is assumed that the relatively resistant cells surviving the first fraction of the dose are sensitized towards translocation induction when receiving the second fraction of the dose.     

Griseofulvin-induced aneuploidy and meiotic delay in male mouse germ cells: detected by using conventional cytogenetics and three-color FISH.

Griseofulvin (GF) was tested in male mouse germ cells for the induction of meiotic delay and aneuploidy. Starved mice were orally treated with 500, 1000 and 2000 mg/kg of GF in corn oil and testes were sampled 22 h later for meiotic delay analysis and chromosome counting in spermatocytes at the second meiotic metaphase (MMII). A dose-related increase in meiotic delay by dose-dependently arresting spermatocytes in first meiotic metaphase (MMI) or/and prolonging interkinesis was observed. Hyperhaploid MMII cells were not significantly increased. Sperm were sampled from the Caudae epididymes 22 days after GF-treatment of the males for three-color fluorescence in situ hybridization (FISH). The frequencies of diploidies were 0.01-0.02% in sperm of the solvent control animals and increased dose-dependently to 0.03%, 0.068% and 0.091%, respectively, for 500, 1000 and 2000 mg/kg of GF. The frequencies of disomic sperm were increased significantly above the controls in all GF-treated groups but showed no dose response. The data for individual classes of disomic sperm indicated that MII was more sensitive than MI to GF-induced non-disjunction in male mice. A comparison of the present data from male mice and literature data from female mice suggests that mouse oocytes are more sensitive than mouse spermatocytes to GF-induced meiotic delay and aneuploidy.     

Studies on the pathogenicity of three host races of Rotylenchulus reniformis on castor

Three races (race 2, 3 and 4) of Rotylenchulus reniformis were identified using host differential from 20 populations collected from Aligarh, India. Pathogenicity tests were conducted using 250, 500, 1000, 2000, 4000 and 8000 immature females/kg soil of three races of R. reniformis on castor. The threshold inoculum levels of Race-2, Race-3 and Race-4 on castor were 500, 1000 and 2000 immature females/kg soil, respectively. Out of these three races present in Aligarh district, Race-2 was found more pathogenic followed by Race-3 and Race-4. Infected plants with all the three races of reniform nematode showed stunting, necrosis, leaf shedding and growth reduction at and above the inoculum threshold level of each race. Reduction in castor growth was directly proportional to the inoculum level of R.reniformis. The rate of nematode multiplication was density dependent.     

Studies on mutagen-sensitive strains of Drosophila melanogaster. VII. Effects of repair deficiency in males on X-ray-induced sex-linked recessive lethals in spermatozoa

The response of mature spermatozoa to the X-ray induction (500 R and 3000 R) of sex-linked recessive lethals was studied in Drosophila melanogaster males known to be deficient in excision- or post-replication repair of UV damage in somatic cells. The results show that the induced frequencies of recessive lethals in the excision-repair-deficient males (mei-9a and mei-9L1) are similar to those in the appropriate repair-proficient males (mei+ and Berlin-K). However, in the post-replication-repair-deficient males (w mus(1)101D1), these frequencies are significantly lower than in the comparable repair-proficient males (w) after 500 R, but not after 3000 R.     

Translocation yield from the mouse spermatogonial stem cell following fractionated X-ray treatments: influence of unequal fraction size and of increasing fractionation interval

(1) The genetic response of the mouse spermatogonial stem cell to a high dose of X-rays given in two unequal fractions 24 h apart can be dependent upon the order in which the two fractions are given. When 1000 R was administered as 100 R followed by 900 R the recovered translocation yield (22%) was similar to that which can be obtained by extrapolation from lower doses and also to that of a 500 + 500 R 24 h fractionation. By contrast, when the 900 R preceded the 100 R the response was much lower (7.4%), yet still greater than that produced by a single 1000 R treatment (4.5%). The same order of effectiveness was observed for length of sterile period. (2) The sub-additive translocation yields previously obtained with 800 R treatments given in fractions of 500 R and 300 R at intervals of 3-12 days were found to be maintained with intervals up to at least 15 days but additivity was regained by the end of the third week. Sterile period data indicated that with these intervals the germinal epithelium had recovered sufficiently from the first fraction for spermatogenesis to restart before the second fraction was given. (3) It is concluded from the two experiments that (a) 24 h after a radiation exposure the surviving stem cells are more sensitive than formerly both to killing and genetic damage, (b) at this time they are no longer heterogeneous in their radiosensitivities, so that increasing yields of genetic damage may be obtained with increasing dose i.e. there is no fall in yield at higher doses, (c) the change in sensitivity could be a consequence of a synchronization to a sensitive stage in a cell cycle, or to a transitional phase preparatory to entering a different cell cycle. (d) to achieve rapid repopulation of the germinal epithelium the surviving stem cells are stimulated to enter a shorter cell cycle and this is the cause of the sub-additive translocation yields with fractionation intervals of 3-15 days, (e) the recommencement of spermatogenesis is associated with the reestablishment of the heterogeneity in radiosensitivity among the stem cells. At this time additive translocation yields can again be recovered.     

Effects of a chromosome-3 mutator gene on radiation-induced mutability in Drosophila melanogaster females

A series of X-irradiation experiments was carried out using Drosophila melanogaster females homozygous for a third chromosome mutator gene and females which had a similar genetic background except that the mutator-bearing third chromosomes were substituted by normal wild-type chromosomes. The mutator females had been previously shown by Gold and Green to manifest a higher level of radiation-induced mutability (as measured by the X-ray-induction of sex-linked recessive lethals) in their pre-meiotic germ cells compared to normal females at an exposure of 100 R. In the presence work, the sensitivity of the pre-meiotic germ cells of mutator and normal females to the X-ray induction (2000 R) of sex-linked recessive lethals was studied. In addition, experiments were conducted to examine the sensitivity of the immature (stage 7; prophase I of meiosis) oocytes of both kinds of females to the induction of dominant lethals, X-linked recessive lethals and X-chromosome losses. The result show that in pre-meiotic germ cells, the frequencies of radiation-induced recessive lethals are similar in both kinds of females. However, the proportion of these mutations that occur in clusters of size 3 and higher, is higher in mutator than in normal females. In stage-7 oocytes, the frequencies of radiation-induced dominant lethals and sex-linked recessive lethals were similar in both kinds of females. The X-loss frequencies however, were consistently higher in mutator females although statistical significance was obtained only at higher exposures (3000 and 3750 R) and not at lower ones (750-2250 R). Possible reasons for the discrepancy between the present results and those of Gold and Green with respect to pre-meiotic germ cells are discussed.     

Specific-locus mutation response to unequal, 1 + 9 Gy X-ray fractionations at 24-h and 4-day fraction intervals

The specific-locus mutation frequency obtained from mouse spermatogonial stem cells following unequal, 1 + 9 Gy X-ray fractionation with a 24-h fractionation interval is low, and consistent with the two fractions acting additively. The response is therefore markedly different from the augmented mutation frequencies obtained with 500 + 500 R and 100 + 500 R, 24-h fractionations. The lower yield compared with the 100 + 500 R response also indicates a clear difference from the translocation data which demonstrate increases in yield with increasing second dose over the same dose range. The decline in specific locus mutation yield with the increase in the second dose from 500 R to 9 Gy suggests that the stem cells surviving the first fraction are heterogeneous in their sensitivities to this class of genetic damage. A similar, additive specific locus mutation frequency is obtained with unequal, 1 + 9 Gy X-irradiation when the interval between fractions is 4 days. This is consistent with 500 + 500 R, 4-day and 7-day interval responses obtained previously but again differs from the sub-additive translocation responses obtained with such X-ray fractionation. Taken together with the data from previous studies the present results suggest that (1) 24 h after the first fraction, (a) the surviving stem cell have two components; survivors of the formerly radiosensitive, cycling component of the normal stem cell population and the formerly radioresistant, G0 or arrested G1 cells, which are being 'triggered' into a rapid cell cycle to achieve repopulation of the testis; (b) these two components are of near-equal sensitivity to translocation induction and cell killing, hence the additive translocation yields with equal X-ray fractionations and yields consistent with those extrapolated from lower doses with higher, unequal fractionations, e.g. 1 + 7 Gy, 1 + 9 Gy; but (c) the formerly radioresistant, triggered component is much more sensitive than the surviving cycling component to specific locus mutation and cell killing, hence the augmented mutation response with 500 + 500 R fractionation and the drop in yield with 1 + 9 Gy compared with 100 + 500 R X-irradiation.(ABSTRACT TRUNCATED AT 400 WORDS)     

Mutational changes of quantitative morphological traits in the convict cichlid (Cichlasoma nigrofasciatum Guenther) after irradiation of parental spermatogonia and oogonia with different doses of X-rays

10 continuous and 6 discrete variables of body dimensions as well as 16 proportions calculated from them were determined in postirradiation F1 convict cichlid fish (Cichlasoma nigrofasciatum) which were derived from spermatogonia and oogonia both exposed to either 0 (controls), 250, 500, 750, 1000, or 2000 R of X-rays. While no interactions with sex or radiation dose were found, clear dosage effects and some sex effects occurred. The highest response for almost all quantitative traits examined as well as for variants of coloration or of vertebral aberrations appeared after 250 R to both parental germ cells. The lowest effectiveness was observed for almost all characters under investigation after 0 and 500 R. Accordingly, the largest difference between the 6 different treatment groups was found between 250 and 500 R, followed by 250 and 0, 250 and 1000, and 250 and 750 R to parental gonia. The present finding on the alterations of quantitative morphological traits completely corresponds to the changes of intraspecific aggressiveness as found for the same individual fish in a previous study thus confirming the complete congruence of the mutational response of quantitative morphological characters with that of behavioral traits to ionizing radiation.     

Forecasting yield of corn,Zea mays infested with corn leaf aphid,Rhopalosiphum maidis

Abstract: Corn leaf aphid, Rhopalosiphum maidis (Fitch) is one of the most serious corn pests in Egypt. The aim of the present study is to assess the corn yield losses and to build forecasting models of yield related with aphid infestation. Number of aphids/plant were determined during corn growth stages 10 leaves (V10); tasseling (VT); ripening 2 (R2) and ripening 4 (R4). At the end of growing season, yield losses were estimated. Results revealed that infestation with aphids through V10–VT caused 28.14% yield losses at average aphid density 818 aphid/plant. Infestation through R2–R4 caused 16.28% yield losses at average aphid density 1038 aphids/plant. Percentages of yield losses of corn ears through V10–R4 were 14.66, 22.9, 35.28 and 36.03% at average aphid density of 100, 1000, 2000 and 3000 aphids/plant. As ear yield negatively correlated with aphid density, regression analysis was used to construct forecasting yield models. Statistical analysis showed that simple linear and logarithmic models provided a good fit to the data and also indicated that the two models are similar in prediction of ear yield. These constructed models were diagnostic checked using new data. The checking revealed that the linear model is stable and valid where insignificant difference was observed between predicted and actual yield.     

Biochemical characterization of six trisomics of grain sorghum,Sorghum bicolor (L.) Moench

To determine protein differences of grain sorghum disomics and trisomics, we analyzed leaf extracts from six trisomics and a disomic control by disc gel, gel isoelectric focusing, and SDS gel electrophoresis. Based on the number and position of protein bands revealed by Commassie blue staining, the disomic control could be differentiated from the trisomics, and trisomics could be shown to differ among themselves in most cases. SDS gels revealed the most protein bands, followed by isoelectric focusing and disc gel. However, disc gel electrophoresis was the simplest technique of the three and was just as effective in identifying trisomics and differentiating trisomics from the disomic control.Contribution 1596-j, Department of Agronomy, and 182-j, Department of Biochemistry, Kansas State University, Manhattan, Kansas.     

Irradiation increases superoxide dismutase in rat intestinal smooth muscle

We investigated whether X-irradiation could induce the enzyme superoxide dismutase (SOD) in intestinal muscle. Groups of rats received abdominal irradiation and the time course and dose response for SOD activity determined. Jejunal smooth muscle homogenates were analyzed for the activities of copper/zinc (CuZn) and manganese (Mn) SOD activity and for a mitochondrial marker enzyme, citrate synthase. A progressive rise in Mn SOD activity occurred at 20, 46, and 72 h after 1500 R. No significant changes in Cu-Zn SOD activity occurred at any time after 1500 R. At 20 h after 250 R of X-irradiation, Mn SOD activity increased but no further increase occurred at higher irradiation exposures. At the same time, CuZn SOD activity at 20 h after irradiation was greater than controls only at an exposure of 1000 R (p less than 0.05). Using Western blotting, we were able to clearly demonstrate an increase in immunoreactive Mn SOD protein in muscle samples 20 h after 1500 R. The rise in Mn SOD is not simply due to increase in mitochondrial numbers or increase in all mitochondrial enzyme activities because activity of the mitochondrial marker enzyme citrate synthase was decreased after X-irradiation. Transmission electron microscopic studies demonstrated damage to mitochondria after a dose of 3000 R. The data yield evidence that free radicals play a role in irradiation-induced intestinal smooth muscle injury.     

Macrofilaricidal and microfilaricidal effects of Neurolaena lobata, a Guatemalan medicinal plant, on Brugia pahangi

Twelve extracts of 11 Guatemalan medicinal plants were initially screened in vitro for potential macrofilaricidal activity against Brugia pahangi, a lymphatic dwelling filarial worm, using concentrations from 125 to 1000 microg ml(-1) of each extract that could be dissolved in the culture medium. Of 12 extracts used, the ethanol extract of leaves of Neurolaena lobata showed the strongest activity against the motility of adult worms. Subsequently, the extract of N. lobata was extensively examined in vitro for macro- and micro-filaricidal effects using a series of concentrations of 500, 250, 100, 50 and 10 microg ml(-1). The effects were assessed by worm motility, microfilarial release by female worms and a MTT assay. The effect on the motility of adult worms was observed in a concentration- and time-dependent manner. The time required to stop motility of both sexes of adult worms was 6 h at 500 microg ml(-1), 24 h at 250 microg ml(-1), and 3 days for females and 4 days for males at 100 microg ml(-1). The movement of females ceased at 4 days at a concentration of 50 microg ml(-1) whereas the motility of males was only reduced. The loss of worm's viability was confirmed by the MTT assay and was similar to the motility results. These concentrations, including 10 microg ml(-1), prevented microfilarial release by females in a concentration- and time-dependent manner. Concentrations higher than 100 microg ml(-1) even induced mortality of the microfilariae. The present study suggested that the ethanol extract of Neurolaena lobata has potential macro- and micro-filaricidal activities.     

Die Reaktion des neurosekretorischen Zwischenhirn-Hypophysensystems beim Meerschweinchen nach Röntgenbestrahlung (200 kV)

Zusammenfassung Der Kopf von insgesamt 35 männlichen und weiblichen Meerschweinchen wurde mit Röntgenstrahlen (200 kV) homogen einzeitig bestrahlt. Die Dosen betrugen 500, 1000, 2000 und 3000 R bei einer Dosisleistung von 100 R/min. Nach allen Dosen wird in den neurosekretorischen Kernen des Hypothalamus und im Hypophysenhinterlappen der Bestand an Neurosekret vermindert. Drei Tage nach den Bestrahlungen mit 500 R nimmt das Neurosekret wieder zu und die Menge normalisiert sich bis zum 10. Tag p. irrad. Nach 1000 bis 3000 R sterben die Tiere nach 1 Woche, ohne daß es zur Restitution der Neurosekretbildung kommt.

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The reaction of the neurosecretory hypothalamo-hypophyseal complex in the guinea pig following X-irradiation

Summary The effect of 200 kV X-irradiation on the hypothalamus and hypophysis of 35 (both male and female) guinea pigs was examined 24, 72, 120, and 240 hours after irradiation of the head. Doses of 500, 1,000, 2,000 and 3,000 R were used at 100 R per minute. 24 hours following irradiation at all doses, a decrease of neurosecretory material occurred in the nucleus supraopticus, nucleus paraventricularis, and in the neurohypophysis. 72 hours following irradiation with 500 R, the amount of neurosecretory material increased and normal levels are attained at 10 days. Higher dosages (1,000 to 3,000 R) prevented the increase observed with 500 R at 72 hours; rather, the animals died after 1 week.

     

Genetic analysis of the mitochondrial rho-mutability in Saccharomyces. II. Disomy for chromosome IV and spontaneous rho-mutability

The disomy for chromosome IV in the strains studied led to: reduction in the red pigmentation of ade1 mutant colonies; a decrease in spontaneous rho- mutant frequency, and impairment of sporulation in hybrids descended from disomic parents. The nuclear srm1 mutation decreasing the spontaneous rho- mutability promoted the spontaneous extra chromosome loss in the disomics for chromosome IV. This result suggests a close connection between the spontaneous rho- mutability and mitotic chromosome stability.     

The relative frequency of induced mutations recovered from Drosophila melanogaster gametogenic stages irradiated at different dose rates

Inseminated Drosophila females were exposed to radiation doses of approx. 2000 or 4000 R at dose rates ranging from 8 to 240 000 R/h. The observed frequencies of sex-linked lethal mutations induced in spermatozoa and oogonia suggest that dose-rate effects were absent in spermatozoa over the whole range and in oogonia up to 3000 R/h. There was a consistent increase in oogonial mutation frequency from 3000 to 240 000 R/h. These results are discussed in relation to the well-established dose-rate effects observed in the mouse.     

The contribution of diffusion to the termination of penicillin-induced convulsive activity in the awake rat.

1. Experiments were performed to investigate the contribution of diffusion to the termination of PCN induced convulsive (epileptiform) activity in the awake rat. The basic question was whether convulsive activity terminates at the same critical concentration Ct of PCN independently of the concentration Co of PCN injected into the superficial layer of the foreleg field of the right motor cortex. Different concentrations Co were used, the lowest being 16 IU PCN/0.5 microliters saline, the largest 1000 IU PCN/0.5 microliters saline. The convulsive activity was recorded by means of the electroencephalogram (EEG). 2. Mathematical calculations concerning the diffusion of the substances were performed by using the diffusion equation with a PCN diffusion coefficient D = 3.52 x 10(-4) mm2/s and the tortuosity factor lambda = 1.62. 3. The lowest dose of PCN used was considered as a threshold dose for acute induction of convulsive activity because only 4 out of 9 rats treated with 16 IU PCN developed epileptiform activity. For the range of doses between 16 and 32 IU, the median frequency of convulsive potentials was dose-dependent, and increased from 7.9 to 13.8 pot./min, respectively. For higher doses covering the range between 63 and 1000 IU, there was only an insignificant increase from 19.2 to 22.7 pot./min, respectively. 4. The median period of convulsive activity increased significantly (p less than 0.05 or less than 0.01) and monotonically from 19.5 min in rats treated with 16 IU PCN to 267 min after treatment with 1000 IU PCN. 5. At the end of the convulsive activity, the median critical concentration Ct within the artificial focus ranged from 1.75 to 1.31 IU/0.5 microliters saline in rats treated with 16 to 125 IU PCN, while after induction of convulsive activity with doses of 250, 500 or 1000 IU PCN, Ct gradually increased to 2.01, 2.83, and 3.75 IU/0.5 microliters, respectively. This means that the brain became less sensitive for PCN the longer the epileptiform activity lasted. 6. It is concluded that during long-lasting convulsive activity, the brain takes advantage of its plastic properties by eliciting counteracting mechanisms. These self-protective factors may utilize neuronal networks which needs some time to become effective in the control of the cortical activity.