Ecdysteroids in adult males and females of Drosophila melanogaster

Ecdysteroid titres in whole flies and different tissues of adult male and female Drosophila were determined at various times after eclosion using a radioimmunoassay. The ecdysteroid titre decreased as the flies matured after eclosion. The differences in titre between males and females can be accounted for by their difference in body weight. The ecdysteroids were found to be distributed throughout several tissues. At eclosion not all of the ecdysteroid complement present could be accounted for by that found localised in tissues. After maturation of the flies the ecdysteroids in various tissues can account for the majority of that detected in whole-fly extracts. Ecdysteroids were produced during in vitro culture of various tissues, but the quantities detected were low by comparison with ring glands of wandering 3rd-instar larvae. Neither the ovaries nor the abdominal body walls (fat body) seem to be a major source of hormone, and they are only able to convert minute quantities of ecdysone to the biologically active form, 20-hydroxyecdysone, in vitro. The amounts of 20-hydroxyecdysone present were measured using high performance liquid chromatography and radioimmunoassay. We tentatively suggest that the differential experession of the yolk-protein-genes in the fat bodies of males and females does not result from differences in hormone titres between them.     

Taste responses of Drosophila melanogaster

The amounts of sugar solution consumed by Drosophila melanogaster flies were determined. Starved and desiccated flies of a wild type strain (QA) consume 7?9 × 10^?2 λ of a 0.3 M sucrose solution per fly during the first hour and less later. They consume more of the 0.3 M sucrose solution than of the more diluted and the more concentrated solutions. In preference-aversion tests the flies discriminated between water and various sugar solutions, and between different sugar concentrations. Contrary to other fly species these flies did not prefer 0.05 M fructose over 0.05 M glucose. 0.3–0.5 M NaCl added to 0.1 M sucrose turned a preference over 0.01 sucrose into an aversion. A mutant, Lot-94, selected for its increased consumption of a 1 M NaCl solution was found to consume more of all test solutions. The amount of NaCl that had to be added to 0.1 M sucrose to turn the preference over 0.01 M sucrose by the mutant flies into aversion was not different from that found for the wild type flies.     

Female courtship summation in Drosophila melanogaster

To examine the concept of female courtship summation in Drosophila melanogaster, two experiments previously reported in the literature, the first involving repeated matings of females and the second progressive removal of the males' wings, were repeated. The present results do not convincingly support the concept of female summation of stimuli provided by the males' courtship. The results of the first experiment also refute the idea that low male courtship intensity leads to long courtships. These results also fail to support an earlier suggestion that females summate the sine rather than pulse song component of the males' wing vibration. Instead, the variation in courtship duration appeared to result from the inverse hyperbolic relationship between the male latency to courtship and the subsequent courtship duration. Thus short male latencies led to longer courtship durations. This is interpreted as resulting from a female latency period during which the female is too agitated to receive the male's courtship, and after which she mates upon recognizing the male as conspecific. In addition, very long courtships largely resulted from additional agitation of the female by the male's courtship. Long courtships therefore appear to be an artifact of the experimental situation and the established concept of female courtship summation, which is supposed to explain them, is unnecessary. The implications of this conclusion are discussed.     

Ovarian yolk-protein synthesis in Drosophila melanogaster

The ovaries and fat bodies of Drosophila melanogaster adult females both synthesize yolk polypeptides. In a series of experiments it has been shown that the ovaries become competent to mature in an adult male host, where only ovarian synthesis occurs, very early in metamorphosis and synthesis of yolk polypeptides begins in a time-dependent sequence related to the age of the ovary when transplanted. Maturation of ovaries occurs prior to eclosion when they are transplanted to an earlier developmental stage showing that neither the event of eclosion not the adult environment is essential in triggering yolk-polypeptide synthesis by the ovary. When metamorphosing ovaries are transplanted to a female host they take up host yolk polypeptides from the haemolymph, but this does not lead to the implanted ovary developing substantially better than in a male host where only synthesis by the ovary can occur. The regulation of ovarian yolk-polypeptide synthesis therefore appears to be autonomous to the ovary itself. There may be a trigger early in metamorphosis which induces competence in the ovary so that it subsequently initiates yolk-polypeptide gene expression at eclosion.     

MMS sensitive strains in Drosophila melanogaster

Larval populations of Drosophila melanogaster were treated with the monofunctional alkylating agent methyl methyl methanesulfonate (MMS). Four different stocks were tested. The method methyl used permits the direct registration of MMS-induced lethality of every genotype present in the treated population. Up to 5-fold differences in sensitivity between different genotypes were obeserved. Larvae of the wild-type strain. Antibes, which was reported to have increased UV sensitivity in embryonic stages, were MMS sensitive. In another stock, marked with spa^pol, female larvae were about twice as sensitive as male larvae.     

Drosopterins in phototaxis-selected strains of Drosophila melanogaster

The age-dependent drosopterin concentration was investigated in phototaxis-selected strains (Röko, Röpo, Röne, Stab) of D. melanogaster using extracted head homogenates. The results are compared to the wild stock derived strain Plus. The drosopterin concentration increases with the age of the flies. Females have larger concentrations than males. There are no differences in the drosopterin concentrations among the phototaxis-selected strains. Plus shows a higher drosopterin content than all other strains. This difference might be based on the genetical background, which is different in these strains. The data indicate that the phototactic behaviour is not related to the drosopterin concentration of the eyes.     

Drosophila melanogaster deoxyribonucleoside kinase activates gemcitabine

Drosophila melanogaster multisubstrate deoxyribonucleoside kinase (Dm-dNK) can additionally sensitize human cancer cell lines towards the anti-cancer drug gemcitabine. We show that this property is based on the Dm-dNK ability to efficiently phosphorylate gemcitabine. The 2.2 Å resolution structure of Dm-dNK in complex with gemcitabine shows that the residues Tyr70 and Arg105 play a crucial role in the firm positioning of gemcitabine by extra interactions made by the fluoride atoms. This explains why gemcitabine is a good substrate for Dm-dNK.     

A mating plug protein reduces early female remating in Drosophila melanogaster

Mating plugs are formed within the female reproductive tract during mating from male ejaculate constituents or even from male genitalia themselves. Across species, mating plugs have roles in sperm storage and the prevention of female remating. In the fruitfly Drosophila melanogaster, accessory gland proteins such as the sex peptide are known to reduce female remating, however this effect can take some time to establish, hence other ejaculate components must also be involved. We hypothesised a role for the PEBII mating plug protein in the prevention of early female remating. Using RNA interference we produced PEBII knockdown males. We found that these males were significantly less able to prevent female remating in the 4 h following mating. The mating plugs produced by PEBII knockdown males also showed lower levels of autofluorescence in the first 10 min after the start of mating, suggesting they differed in composition to those of control males. Reduced levels of PEBII had no effect, however, on fecundity, progeny production or egg-adult viability in the first 24 after mating, suggesting there were no short-term effects of PEB II on sperm transfer, storage or use. Our results show that PEBII has a subtle but significant role in the prevention of early female remating.     

Mutagenicity tests with metrifonate in Drosophila melanogaster

The organophosphorus insecticide metrifonate (O,O-dimethyl(1-hydroxy-2,2,2-trichloroethyl)phosphonate), also known as trichlorfon or Dipterex ®, was tested for its ability to induce sex-linked and autosomal recessive lethal mutations in Drosophila melanogaster. There was no evidence of an increase in the frequency of lethal mutations after feeding adult males with metrifonate, but the high toxicity of the compound meant that only low concentrations could be used in this test system.     

Visual signals in the courtship of Drosophila melanogaster

Drosophila melanogaster carrying either of the mutations sev or dipp⁶ show defective phototactic behaviour owing to deficiencies in the processing of visual information perceived by the central retinula cells (R7, R8). Mutant females show increased time to mating because the deficient visual input via this subsystem has an inhibitory effect on female receptivity. Similarly, deficient input through the peripheral retinula cells (R1–R6) also makes females sexually unreceptive. Thus females require appropriate visual stimulation through both subsystems to become maximally sexually receptive. One major source of this stimulation is the red eye of the male.     

An operating cost of learning in Drosophila melanogaster

Although the fitness benefits of learning are well understood, we know little about its costs; yet both are essential to understand the evolution of animal learning. We tested the hypothesis that learning has an operating cost, such that an animal repeatedly forced to use its learning ability would show a reduction in some fitness component(s), relative to an animal of the same genotype that does not have to learn. Five ‘High-learning’ lines of Drosophila melanogaster, which had been selected for improved learning ability, were exposed to 12 consecutive 48-h cycles of alternating conditioning treatment under mild nutritional stress. Their learning score first increased, reaching a maximum around day 12 (i.e. the sixth conditioning cycle), and then progressively declined. These changes were not due to ageing, as they were not observed in flies from the same lines maintained under standard conditions. From around day 12, the productivity (egg-laying rate) of the flies in the conditioning treatment became progressively reduced, relative to flies from the same lines not exposed to conditioning, but otherwise kept under the same food-limited conditions. This reduction in productivity was not observed when these treatments were applied to five ‘Low-learning’ lines, which had not been exposed to selection, and which show no detectable response to conditioning under our experimental conditions. Furthermore, exposure to repeated cycles of conditioning revealed an apparent trade-off between the learning score and productivity among the ‘High-learning’ lines. These results indicate an operating cost of learning, paid only by genotypes that show learning, rather than general effects of stress caused by the conditioning regime. Potential proximate explanations include (1) the impairment of oviposition decisions caused by the accumulation of memory interference and (2) energy costs of collecting, processing and storing information.     

Evolutionary significance of courtship conditioning in Drosophila melanogaster

Males of the fly Drosophila melanogaster initially court mated, unreceptive females but later develop an avoidance reaction against them and even become temporally unresponsive to virgin females. This conditioned inhibition has been described as an associative process, the conditioned stimulus being a mixture of pheromones on the female's cuticle. To assess the evolutionary significance of courtship conditioning we recorded and analysed the male's behaviour during the conditioning process. The time traces of individual males were marked by an abrupt behavioural change. The time he spent courting suddenly decreased, and the frequency of ‘turn-away’ events at the same time sharply increased. Thus, the gradual decline of courtship observed as a group average does not reflect a slow change in motivation of the individual male but rather the interindividual differences of an active, experience-guided all-or-none decision to stop courting and to avoid the female. Three recently collected D. melanogaster strains were each maintained under two distinct mating conditions. Males were kept together with females for either ca. 2 weeks or 18 h. After 21 generations males of the two regimes differed markedly in their behaviour towards mated females. With long interaction periods between males and females, selection favoured courtship conditioning, while with short periods no such selection was observed. Slowly recovering receptivity of mated females may be needed for the maintenance of courtship conditioning. Courtship conditioning in D. melanogaster appears to be a fitness-relevant behaviour adapted to high-density populations with females mating a second time.     

Visual learning behaviour in Drosophila melanogaster wildtype AS

Visual learning in Drosophila populations is characterized by a fast rate of acquisition, and conditioning indices near 0.3 are obtained. Heterogeneity of the populations with respect to this behaviour is small. Reversed conditioning is possible. Prolonged extinction procedures fail to reduce the conditioning index to zero. Memory lasts for a minimum of 14 hours.     

Cuticle secretion during larval growth in Drosophila melanogaster

The moulting cycle and growth of the larval integument of Drosophila melanogaster has been studied by light and electron microscopy. Growth during the first, second and third larval instars is accompanied by 3.0-, 3.4- and 3.7-fold increases in surface area, respectively. Growth in surface area occurs continuously during the larval stages, with no detectable relationship to the moulting cycle. Measurements of the thickness of the cuticular layers show that the endocuticle grows in thickness by apposition and in surface area by stretching. The pre-apolytic epicuticle remains at fairly constant thickness during the increase in surface area, indicating that it grows by intussusception of new components. Post-apolytic epicuticle becomes thinner and increases in surface area by stretching. The epicuticle and pre-ecdysial endocuticle are traversed by filaments, but these do not penetrate the endocuticle secreted after ecdysis. We suggest that the filaments transport breakdown products from the old cuticle inward to the epidermis for reutilization. The growth and deposition of cuticle in two larval growth mutants, lethal (2) giant larvae and Chubby Tubby, involves mechanisms similar to those found in wild-type larvae, but in Chubby Tubby the endocuticle contains inclusions which are ultrastructurally similar to dense epicuticle.     

The stability of Drosophila melanogaster courtship across matings

Sex effects on behaviour were investigated by repeatedly mating the same male and female Drosophila melanogaster. Male influence is mainly on percentage wing vibration while females largely determine wing vibration duration and bout number. This courtship pattern is stable across matings involving the same individuals.     

Molecular characterization of choline acetyltransferase from Drosophila melanogaster

Purified acetyl-CoA: choline O-acetyltransferase (EC 2.3.1.6) from Drosophila melanogaster has been shown to contain two major polypeptides of 67 and 54 K Daltons. However, all enzyme activity is found in a single molecular weight form of approx 67 K Daltons as determined by sucrose gradient sedimentation and molecular exclusion chromatography. The latter showed both the 67 and 54 K Dalton polypeptides on polyacrylamide gel electrophoresis in sodium lauryl sulfate (10% acrylamide). Analysis of purified choline acetyltransferase on polyacrylamide gel electrophoresis in sodium lauryl sulfate (15% acrylamide) revealed the presence of an additional polypeptide at 13 K Daltons. Tryptic-peptide maps of the 67, 54 and 13 K Dalton components showed all three to be structurally related. In addition to several common tryptic peptides, the 13 K Dalton polypeptide contained three tryptic-peptides that were also found in the 67 K Dalton polypeptide, but were absent from the 54 K Dalton polypeptide. This evidence suggests that native Drosophila choline acetyltransferase may exist in two forms, one a single polypeptide chain with a molecular weight of 67 K Daltons and the other consisting of two noncovalently bound polypeptide chains with molecular weights of 54 and 13 K Daltons. The latter form is the major one isolated and may be generated by limited proteolysis of the single chain 67 K Dalton form.