Arrested development of the rabbit stomach worm Obeliscoides cuniculi: varied responses to cold treatment by the offspring produced throughout the course of a single infection

Arrested development of the rabbit stomach worm Obeliscoides cuniculi: varied responses to cold treatment by the offspring produced throughout the course of a single infection. International Journal for Parasitology16: 55–61. In an experimental model, rabbits were inoculated with cold treated L3s (parental generation) to establish infections with a high proportion of arrested larvae. Over 234 days, ten batches of L3s (offspring) were derived sequentially from ten pairs of rabbits. Samples of L3s (offspring) were cold treated for various periods (13–74 days) and inoculated into test-rabbits to determine cold treatment time for maximum arrest (CTTMA). As parental infections proceeded, the CTTMA varied from batch to batch of offspring. Their propensity for arrest also varied, but no correlation was found between duration of infection by parents and propensity for arrest by offspring. Three additional rabbits were treated early in their infections with thiabendazole. Offspring larvae subsequently derived from these rabbits exhibited significantly more arrest than those from untreated hosts. The model shows that, in natural infections, overwintering arrested larvae could transmit genes for arrest to their spring progeny but that such genes would remain unexpressed until acted on by the correct environmental stimulus.     

Arrested development of the rabbit stomach worm Obeliscoides cuniculi: resumption of development of arrested larvae throughout the course of a single infection

Arrested development of the rabbit stomach worm Obeliscoides cuniculi: resumption of development of arrested larvae throughout the course of a single infection. International Journal for Parasitology 16: 47–54. Twenty rabbits were inoculated once with cold treated L3s of Obeliscoides cuniculi with a high propensity for arrest and killed in pairs at intervals over 234 days post-infection. An exponential decline in numbers of adult worms and arrested larvae was observed. Arrested larvae resumed development at a rate seven times greater than adults were lost and appeared to replenish the adult population. More male than female arrested larvae were intially established in rabbits but males resumed development at a greater rate than females. Egg and larval production were correlated with the number of female worms present at any one time. In a subsidiary study, arrested larvae were recovered from one rabbit and transferred sequentially through two further rabbits. These larvae resumed development at the same rate as larvae in the 20 rabbits. Both studies suggest that resumption of development is an intrinsic characteristic of the larvae themselves and may be genetically controlled.     

Method For Probing Cells In Radiation-Induced G2Arrest: Demonstration of Potentially Lethal Damage Repair

Abstract. Chinese hamster ovary cells were arrested in the G₂ phase of the cell cycle by X-irradiation. When subsequently treated with 5 mM caffeine the arrested population progressed into mitosis as a synchronous cohort where it was harvested by mitotic cell selection. This procedure provides a means to isolate cell populations treated in G₂, for the investigation of G₂ arrest. Comparisons were made of the number of cells retrieved from G₂ arrest with the number suffering arrest, as determined by flow cytometry and by matrix algebraic simulations of irradiated cell progression. the retrieved population was not significantly less than expected for doses up to 3.5 Gy, indicating that the retrieval process does not favour the isolation of any population subset below this dose. Cell populations retrieved from arrest at varying intervals (0-3 h) after irradiation (0-3.5 Gy) showed an increase in survival with increase in interval, consistent with repair of potentially lethal damage. the repair curves (surviving fraction us time) were each described by a single exponential. G₂ cells that were brought to mitosis without a period of arrest exhibited the same radiation response as cells irradiated in mitosis.     

The evolution of plasticity of dauer larva developmental arrest in the nematode Caenorhabditis elegans

Organisms can end up in unfavourable conditions and to survive this they have evolved various strategies. Some organisms, including nematodes, survive unfavourable conditions by undergoing developmental arrest. The model nematode Caenorhabditis elegans has a developmental choice between two larval forms, and it chooses to develop into the arrested dauer larva form in unfavourable conditions (specifically, a lack of food and high population density, indicated by the concentration of a pheromone). Wild C. elegans isolates vary extensively in their dauer larva arrest phenotypes, and this prompts the question of what selective pressures maintain such phenotypic diversity? To investigate this we grew C. elegans in four different environments, consisting of different combinations of cues that can induce dauer larva development: two combinations of food concentration (high and low) in the presence or absence of a dauer larva-inducing pheromone. Five generations of artificial selection of dauer larvae resulted in an overall increase in dauer larva formation in most selection regimes. The presence of pheromone in the environment selected for twice the number of dauer larvae, compared with environments not containing pheromone. Further, only a high food concentration environment containing pheromone increased the plasticity of dauer larva formation. These evolutionary responses also affected the timing of the worms’ reproduction. Overall, these results give an insight into the environments that can select for different plasticities of C. elegans dauer larva arrest phenotypes, suggesting that different combinations of environmental cues can select for the diversity of phenotypically plastic responses seen in C. elegans.     

Method for probing cells in radiation-induced G2 arrest: demonstration of potentially lethal damage repair

Chinese hamster ovary cells were arrested in the G2 phase of the cell cycle by X-irradiation. When subsequently treated with 5 mM caffeine the arrested population progressed into mitosis as a synchronous cohort where it was harvested by mitotic cell selection. This procedure provides a means to isolate cell populations treated in G2, for the investigation of G2 arrest. Comparisons were made of the number of cells retrieved from G2 arrest with the number suffering arrest, as determined by flow cytometry and by matrix algebraic simulations of irradiated cell progression. The retrieved population was not significantly less than expected for doses up to 3.5 Gy, indicating that the retrieval process does not favour the isolation of any population subset below this dose. Cell populations retrieved from arrest at varying intervals (0-3 h) after irradiation (0-3.5 Gy) showed an increase in survival with increase in interval, consistent with repair of potentially lethal damage. The repair curves (surviving fraction vs time) were each described by a single exponential. G2 cells that were brought to mitosis without a period of arrest exhibited the same radiation response as cells irradiated in mitosis.     

Ambulatory dispersal in <Emphasis Type="Italic">Tetranychus urticae</Emphasis>: an artificial selection experiment on propensity to disperse yields no response

Dispersal to new hosts is an important process for an invasive herbivore, such as the two-spotted spider mite. A recent study, using artificial selection experiments, has suggested that genetic variation and genetic trade-offs are present for propensity to disperse in this species. However, due to the experimental setup alternative explanations for the response to selection could not be ruled out. Using an altered setup, we investigated whether the propensity for ambulatory dispersal differs genetically between individuals and whether genetic correlations with life-history traits exist. Upward and downward selection on propensity to leave the colony was performed for seven generations in four replicate artificial selection experiments and the results were compared to control lines. No consistent responses to selection were found and no significant effect on life-history traits (oviposition rate, juvenile survival, development rate and number of adult offspring) or sex ratio was present across the replicates. The data suggest that our base population of spider mites harbours at best a low amount of additive genetic variation for this behaviour.     

Methylxanthine inhibition of neurosecretion in the brain and corpus cardiacum of Cecropia

The fine-structure of the median neurosecretory cells and corpora cardiaca of the Cecropia silkmoth during the first 7 days after transfer from cold conditions to room temperature was compared to that of similar animals whose development was arrested with aminophylline. The major difference observed was the failure of the intrinsic secretory cells of the corpus cardiacum to degenerate in the arrested animals. This failure to degenerate coincides with the expected period of brain hormone release. After long periods of arrest, the medial neurosecretory cells and their axons became distended with neurosecretory granules. The significance of these observations in the initiation of adult development is discussed.     

Correlational selection on life history traits in the pitcher-plant mosquito

We examined the short-term effects of genetic architecture on the response to correlational selection in the pitcher-plant mosquito, Wyeomyia smithii. Previously, a positive phenotypic and genetic correlation between development time and propensity to diapause had been identified. Correlational selection consisted of all four combinations of (1) fast and slow development time and (2) propensity to diapause and not diapause, 3 replicates per selection regime, for 1 to 3 generations. Response to selection in the direction of the correlation was rapid and generally consistent with the direction of selection while response orthogonal to the correlation was inconsistent and frequently in a direction opposite to the direction of selection. We conclude that genetic architecture can have a significant, and quite asymmetrical and non-directional, effect on the response to correlational selection.     

Lethal response of HeLa cells to x-irradiation in the latter part of the generation cycle.

The age-response for the killing of HeLa S3 cells by X-rays during the latter part of the generation cycle has been examined in detail. As synchronous cells move from the G1/S boundary through S phase, the relatively high sensitivity of late G1 cells gradually decreases; minimum sensitivity is reached in mid-S and maintained during the remainder of that phase. The response of cells as they progress from S to the point in G2 at which they are temporarily arrested by radiation (or by inhibitors of protein synthesis) was measured in populations free of both S phase cells and late G2 cells that had passed the arrest point: cells retain their high resistance from early G2 up to the arrest point. The response of G2 cells that have passed the arrest point before being irradiated was examined by exposing randomly growing cultures to X-rays and collecting cells periodically thereafter, as they entered mitosis. Survival values very close to those of sensitive mitotic cells were found in the 2 h period after irradiation during which unarrested cells continued to reach mitosis. Values typical of lateS/early G2 were found only after cells that had been arrested began arriving at mitosis. Thus, HeLa S3 cell undergo an abrupt increase in sensitivity at or near the arrest point. The sensitivity to a second irradiation of cells arrested in G2 by a conditioning X-ray dose increases rapidly in the early part of the arrest period.     

Purines inhibit the development of mouse embryos in vitro

The first cleavage of embryos derived from random-bred, inbred, and hybrid-inbred female mice was not arrested by purines at concentrations as high as 30 microM. Development after the first or second cleavage was arrested by hypoxanthine, adenosine or inosine, but not guanosine. In agreement with previous results, the purine-induced block was reversed when arrested embryos were transferred to purine-free media after 24 h in culture. The cleavage arrest was not due to elevations of cAMP as a result of inhibition of phosphodiesterase activity since similar concentrations of phosphodiesterase inhibitors or dibutyryl cAMP did not block development. Treatment with inhibitors of enzymes that convert IMP to AMP or to GMP did not reverse the hypoxanthine-induced block, thus demonstrating that mitotic arrest is mediated by a mechanism different from the hypoxanthine arrest of meiosis. Thymidine incorporation studies showed that the block did not prevent the onset of DNA synthesis. The results reveal a profound sensitivity to purine inhibition of a cell process that occurs during the first 30 h of mouse embryo development and is necessary for progession through the G2 or M phases of the second or third cleavage.     

Characterization of adriamycin-induced G2 arrest and its abrogation by caffeine in FL-amnion cells with or without p53

We investigated the effect of Adriamycin on FL-amnion (FL) cells. After treatment with the drug, the cells arrested at G2, but we did not detect an increase in the p21 levels. We established a p53-deficient derivative of these cells, in which G2 arrest also occurred after treatment with Adriamycin, suggesting that the arrest we observed in these cells is independent of the p53 pathway. Low doses of Adriamycin (100-200 ng/ml) induced G2 arrest, while late S-phase arrest was observed at high doses (500-1000 ng/ml) in both FL and p53-deficient FL cells. Accumulation of cyclin B1 was detected only in cells arrested at G2, and not in those arrested at S phase, suggesting that the S-phase checkpoint functioned efficiently even in p53-deficient FL cells. In both cell lines, caffeine-induced activation of CDC2 kinase was detected only in cells arrested at G2 and CDC2 kinase-activated cells died exhibiting features of apoptosis. CDC2 kinase activation was inhibited by cycloheximide. Furthermore, cycloheximide inhibited activation of CDK2:cyclin A, which normally precedes CDC2 kinase activation in caffeine-treated cells. These results suggest that p53 and p21 do not have special roles in the S- and G2-phase checkpoints and that CDK2:cyclin A could be the target of the G2-phase DNA damage checkpoint.     

Artificial selection for increased dispersal results in lower fitness

Dispersal often covaries with other traits, and this covariation was shown to have a genetic basis. Here, we wanted to explore to what extent genetic constraints and correlational selection can explain patterns of covariation between dispersal and key life‐history traits—lifespan and reproduction. A prediction from the fitness‐associated dispersal hypothesis was that lower genetic quality is associated with higher dispersal propensity as driven by the benefits of genetic mixing. We wanted to contrast it with a prediction from a different model that individuals putting more emphasis on current rather than future reproduction disperse more, as they are expected to be more risk‐prone and exploratory. However, if dispersal has inherent costs, this will also result in a negative genetic correlation between higher rates of dispersal and some aspects of performance. To explore this issue, we used the dioecious nematode Caenorhabditis remanei and selected for increased and decreased dispersal propensity for 10 generations, followed by five generations of relaxed selection. Dispersal propensity responded to selection, and females from high‐dispersal lines dispersed more than females from low‐dispersal lines. Females selected for increased dispersal propensity produced fewer offspring and were more likely to die from matricide, which is associated with a low physiological condition in Caenorhabditis nematodes. There was no evidence for differences in age‐specific reproductive effort between high‐ and low‐dispersal females. Rather, reproductive output of high‐dispersal females was consistently reduced. We argue that our data provide support for the fitness‐associated dispersal hypothesis.     

Water Deficit in Pea Root Tips: Effects on the Cell Cycle and on the Production of Dehydrin-like Proteins

Dehydrin-like proteins have been detected in nuclei and cytoplasmof meristematic root tip cells from pea seedlings subjectedto slow dehydration at 90% relative humidity for 48 h or more.Evidence was gained from Western blotting and immunocytochemicalexperiments using an antibody raised against the conserved domainof dehydrin proteins. Flow cytometer analysis has shown thatcycling cells of root tip meristems from dehydrated seedlingsare mostly arrested in G2 phase. Other stress treatments thoughtto involve water depletion (osmotic stress, cold treatment)or to modulate cell response to water deficit (abscisic acid)gave less clear-cut results with all treatments lowering theproportion of cells entering the S phase, but without a definiteand persistent arrest in any preferential phase of the cycle.Possible interrelationships between G2 arrest and dehydrin productionare discussed. Cell cycle; dehydrins; flow cytometry; nuclei; pea; Pisum sativum L.; water stress     

Effects of caffeine and aminophylline on adult development of the Cecropia silkmoth

Caffeine and aminophylline were potent inhibitors of adult development in the moth, Hyalophora cecropia, when administered prior to initiation of pharate adult development. A dose of 1 mg/g live weight was 100 per cent effective in bringing about this arrest. Cyasterone and α-ecdysone induced development in arrested animals and arrested dauer pupae. Brain transplant and perfusion experments indicated that synthesis or release of brain hormone was disrupted by these drugs. In addition, the effects of ecdysone and perfusion on arrested animals suggested that methylxanthines may also act at other sites in the sequence of events leading to adult development.     

Elevated level of gas3 gene expression is correlated with G0 growth arrest in human fibroblasts

The gas3/PMP22 gene product is a dual function protein, involved in both peripheral nerve myelination and cell proliferation. gas 3/PMP22 is highly expressed in myelinating Schwann cells and is required for normal PNS development. In addition, a more general function for gas3 is suggested by its expression in non-neural tissues and upregulation by growth arrest in cultured rodent fibroblasts. In the present work, the expression of the gas3 gene has been studied in human fibroblasts. We have confirmed that gas3 mRNA is upregulated when cells are serum starved or grown to high cell density (G0 arrest). When quiescent cells were stimulated by serum or platelet-derived growth factor-BB (PDGF-BB), gas3 mRNA was down regulated. In contrast, we found that the expression of gas3 mRNA was neither upregulated in senescent cells nor in cells arrested in G1 using Lovastatin. Thus, high expression of gas3 is not related to growth inhibition in general, but more probably to the G0 growth arrest state. Furthermore, we found that in two malignant fibrous histiocytoma cell lines, gas3 expression was lower than in normal fibroblasts, suggesting an altered regulation of the gas3 gene in transformed cells.     

Study of the cytolethal distending toxin-induced cell cycle arrest in HeLa cells: involvement of the CDC25 phosphatase

HeLa cells exposed to Escherichia coli cytolethal distending toxins (CDT) arrest their cell cycle at the G2/M transition. We have shown previously that in these cells the CDK1/cyclin B complex is inactive and can be reactivated in vitro using recombinant CDC25 phosphatase. Here we have investigated in vivo the effects of CDC25 on this cell cycle checkpoint. We report that overexpression of CDC25B or CDC25C overrides an established CDT-induced G2 cell cycle arrest and leads the cells to accumulate in an abnormal mitotic stage with condensed chromatin and high CDK1 activity. This effect can be counteracted by coexpression of the WEE1 kinase. In contrast, overexpression of CDC25B or C prior to CDT treatment prevents G2 arrest and allows most of the cells to progress through mitosis with only a low percentage of cells arrested in abnormal mitosis. The implications of these results on the biochemical nature of the CDT-induced cell cycle arrest are discussed.     

Testing alternative methods for purging genetic load using the housefly (Musca domestica L.)

When a population faces long-term inbreeding, artificial selection, in principle, can enhance natural selection processes for purging the exposed genetic load. However, strong purge pressures might actually decrease fitness through the inadvertent fixation of deleterious alleles and allelic combinations. We tested lines of the housefly (Musca domestica L.) for the effectiveness of artificial selection to promote the adaptation to small population size. Specifically, replicate populations were held at average census sizes of 54 for nine generations or 30 for 14 generations while being subjected to artificial selection pressure for increased fitness in overall mating propensity (i.e., the proportion of virgin male–female pairs initiating copulation within 30 min), while also undergoing selection to create differences among lines in multivariate components of courtship performance. In the 14-generation experiment, a subset of the lines were derived from a founder-flush population (i.e., derived from three male–female pairs). In both experiments, we also maintained parallel non-selection lines to assess the potential for natural purging through serial inbreeding alone. Sub-populations derived from a stock newly derived from the wild responded to artificial selection for increased mating propensity, but only in the short-term, with eventual rebounds back to the original levels. Serial inbreeding in these lines simply reduced mating propensity. In sub-populations derived from the same base population, but 36 generations later, both artificial selection and serial inbreeding increased mating propensity, but mainly to restore the level found upon establishment in the laboratory. Founder-flush lines responded as well as the non-bottlenecked controls, so we base our major conclusions on the comparisons between fresh-caught and long-term laboratory stocks. We suggest that the effectiveness of the alternative purge protocols depended upon the amount of genetic load already exposed, such that prolonged periods of relaxed or altered selection pressures of the laboratory rendered a population more responsive to purging protocols.     

甜菜夜蛾虫酰肼抗性品系选育及相对适合度研究

本试验在室内对甜菜夜蛾Spodoptera exigua(Hübner)武汉敏感品系进行了8代群体汰选后分别于9、10、12代采用了单对汰选。结果表明虫酰肼处理后各单对品系后代幼虫存活率发生明显分离,F9(s1)、F10(s2)和F12(s3)代幼虫平均存活率分别为48.34%、11.72%和2.37%,筛选后分别为84.00%、83.33%和7.29%。经过12代汰选后,汰选品系抗性倍数为敏感品系的5.24倍,且单对汰选世代对虫酰肼的抗性发展较快,其抗性倍数分别是群体汰选的1.27、1.67和1.38倍。比较单对汰选世代与群体汰选世代某些生物学特性,显示单对汰选世代并不存在生长发育和生殖不利性,表明在虫酰肼群体汰选中穿插几代单对汰选可以加速抗性品系的选育。汰选品系相对于敏感品系具有0.31的适合度,表明甜菜夜蛾对虫酰肼产生抗性后存在适合度缺陷。     

Rb+ influxes differentiate between growth arrest of cells by different agents

The effect of cell cycle on Rb+ (K+) fluxes was studied in NIH 3T3 mouse fibroblasts. Serum starvation or isoleucine deprivation resulted in cell arrest at an early G1/G0 phase, accompanied by a marked decrease in both ouabain-sensitive and ouabain-resistant Rb+ influx. On the other hand, cells arrested at late G1/G0 phase by hydroxyurea treatment have high ouabain-sensitive and ouabain-resistant Rb+ influx. Butyric acid treatment resulted in cell arrest at an early G1/G0 phase, but in contrast to serum or isoleucine starvation did not decrease Rb+ influxes. It is thus shown that quiescent cells may have Rb+ influx rates as high as that of logarithmically growing cells. The results are consistent with the hypothesis that an increased ion permeability of the cell is initiated at a critical stage in G1/G0 phase, and that butyric acid may arrest the cell beyond that stage.     

Venom is beneficial but not essential for development and survival of Nasonia

1. Parasitoid wasps sting and inject venom into arthropod hosts, which alters host metabolism and development while keeping the host alive for several days, presumably to induce benefits for the parasitoid young. 2. This study investigates the consequences of host envenomation on development and fitness of wasp larvae in the ectoparasitoid Nasonia vitripennis, by comparing wasps reared on live unstung, previously stung, and cold‐killed hosts. Developmental arrest and suppression of host response to larvae are major venom effects that occur in both stung and cold‐killed hosts, but not in unstung hosts, whereas cold‐killed hosts lack venom effects that require a living host. Thus, cold‐killed hosts mimic some of the effects of venom, but not others. 3. Eggs placed on live unstung hosts have significantly higher mortality during development; however, successfully developing wasps from these hosts have similar lifetime fecundity to that of wasps from cold‐killed or stung hosts. Therefore, although venom is beneficial, it is not required for wasp survival. 4. While wasps developing on both cold‐killed and stung hosts have similar fitness levels, rearing multiple generations on cold‐killed hosts results in significant fitness reductions of wasps. 5. It is concluded that the largest benefits of venom are induction of host developmental arrest and suppression of host response to larva (e.g. immune responses), although more subtle benefits may accrue across generations or under stressful conditions.