Morphology of the pupal heart, adult heart, and associated tissues in the fruit fly, Drosophila melanogaster.

The early pupal heart of the fruit fly Drosophila melanogaster has recently been the subject of intense physiological and molecular work, yet it has not been well described, nor has it been compared with the heart of the adult fly. In the work reported here, the hearts of adults and early pupae of D. melanogaster were studied by scanning and transmission electron microscopy and by light microscopy. The hearts of adults and early pupae both consist of a tube of circular striated muscle one cell in thickness. The alary muscles, which suspend the heart, are more delicate in the adult compared to the early pupa. The pericardial cells in both early pupae and adults are connected to the heart by connective tissue radiating from the alary muscles or dorsal diaphragm. We confirm that four major changes occur in the heart during metamorphosis: 1) a conical chamber is formed de novo in the first and second abdominal segments; 2) the adult heart curves to conform to the contour of the abdomen; 3) a layer of longitudinal striated muscle appears on the ventral surface of the heart; 4) a fourth pair of ostia is added to the three already present in the early pupa; and note additionally that 5) the ostia appear as simple openings in the heart of the early pupa but are valve-like in the adult.

The effects of period mutations and light on the activity rhythms of Drosophila melanogaster.

Strains of Drosophila melanogaster homozygous for alleles of the period gene (perO, perL, perS, and per+) were reared for multiple generations either in light:dark cycles (LD), continuous illumination (LL), or chronic darkness (DD). The locomotor activity of adult flies from these cultures was monitored in either LL or DD. Flies that were reared and tested in DD had a lower proportion of individuals with normal circadian rhythms than flies reared in LD or LL and tested in DD. The activity rhythms of DD-reared DD-tested animals, when present, showed phase coherence within two out of seven populations, while 8 out of 10 LL-reared DD-tested showed phase coherence. Flies tested in LL were largely devoid of circadian rhythms regardless of their rearing environment. Ultradian rhythms were more evident under conditions disruptive to circadian rhythmicity, but were observed in the presence and absence of circadian rhythms. The periods of the ultradian rhythms of LL-reared DD-tested and LD-reared DD-tested flies varied significantly among genotypes, while in other rearing and testing regimes, no relationship was found.

Effects of deuterium oxide and temperature on heart rate in Drosophila melanogaster.

A non-intrusive optical technique has been developed to monitor heartbeat in late third-instar Drosophila larvae. Heartbeat in this insect is an oscillation that is not temperature compensated. Deuterium oxide lengthens the period of a number of high and low frequency oscillators and clocks in a variety of organisms. To determine whether deuterium affects heart rate, flies were raised on proteated and deuterated media and their heartbeat was monitored at four temperatures ranging from 18 to 33 degrees C. The rate of heartbeat increased linearly with increasing temperature, and decreased with increasing concentrations of deuterium. There was a significant interaction between temperature and deuterium: the higher the concentration of deuterium oxide the less temperature-sensitive was the heart rate. Raising temperatures also increased the amount of "noise" in the rhythm: signal-to-noise ratio, which characterizes the amount of power in a rhythmic signal, decreased with increasing temperatures. Deuterium oxide had no effect on signal-to-noise ratio.

Mutational analysis of the Drosophila miniature-dusky (m-dy) locus: effects on cell size and circadian rhythms.

A mutational analysis has been performed to explore the function of the Drosophila melanogaster miniature-dusky (m-dy) locus. Mutations at this locus affect wing development, fertility and behavior. The genetic characterization of 13 different mutations suggests that m and dy variants are alleles of a single complex gene. All of these mutations alter wing size, apparently by reducing the volume of individual epidermal cells of the developing wing. In m mutants, epidermal cell boundaries persist in the mature wing, whereas they normally degenerate 1-2 hr after eclosion in wild-type or dy flies. This has permitted the direct visualization of cell size differences among several m mutants. Mutations at the m-dy locus also affect behavioral processes. Three out of nine dy alleles (dyn1, dyn3 and dyn4) lengthen the circadian period of the activity and eclosion rhythms by approximately 1.5 hr. In contrast, m mutants have normal circadian periods, but an abnormally large percentage of individuals express aperiodic bouts of activity. These behavior genetic studies also indicate that an existing "rhythm" mutation known as Andante is an allele of the m-dy locus. The differential effects of certain m-dy mutations on wing and behavioral phenotypes suggest that separable domains of function exist within this locus.

High-resolution analysis of locomotor activity rhythms in disconnected, a visual-system mutant of Drosophila melanogaster.

Free-running locomotor activity and eclosion rhythms of Drosophila melanogaster, mutant at the disconnected (disco) locus, are substantially different from the wild-type phenotype. Initial periodogram analysis revealed little or no rhythmicity (Dushay et al., 1989). We have reanalyzed the locomotor activity data using high-resolution signal analysis (maximum-entropy spectral analysis, or MESA). These analyses, corroborated by autocorrelograms, uncovered significant residual circadian rhythmicity and strong ultradian rhythms in most of the animals tested. In this regard the disco mutants are much like flies expressing mutant alleles of the period gene, as well as wild-type flies reared throughout life in constant darkness. We hypothesize that light normally triggers the coupling of multiple ultradian oscillators into a functional circadian clock and that this process is disrupted in disco flies as a result of the neural lesion.

Further evidence that the circadian clock in Drosophila is a population of coupled ultradian oscillators.

We hypothesize that ultradian oscillators are coupled to yield a composite circadian clock in Drosophila. In such a system, period would be a function of the tightness of coupling of these oscillators, increasing as coupling loosens. Ultradian oscillations would become apparent under weak coupling or in the absence of coupling. A new technique for calculating signal-to-noise ratio (SNR) for biological rhythms to characterize their precision has yielded support for this hypothesis. SNR of rhythms of the allelic series of mutations at the period (per) locus of Drosophila melanogaster were compared. Per(o) was the noisiest, grading through perL, per+, and pers, the least noisy. SNR decreases significantly with increasing period in pers, per+, and perL; per(o) typically has multiple ultradian oscillations and the lowest SNR. At least 70% of perL individuals also exhibit ultradian periodicities.

A model describing the kinetics of mating in Drosophila.

The kinetics of mating in species of Drosophila that copulate for periods of time greater than the duration of experimental observation can be described precisely by an equation that combines second order reaction kinetics with the exponential decay of a lag factor. The two rate constants for this kinetic system were estimated from mating data using a wild-type D. simulans strain, a wild-type D. melanogaster strain, and vestigial mutant D. melanogaster males with wild-type females. The results confirm the applicability of second order kinetics, and demonstrate the usefulness of the model in comparisons of mating rates among strains of flies. In most cases, the model explains more than 95% of the variance in the data, and in addition sheds light on the mechanics of the process of mating.

Effects of female behavior on wing display and courtship of male Drosophila simulans and Drosophila melanogaster.

Corrolations between female rejection behaviors and male wing display were calculated for both Drosophila simulans and Drosophila melanogaster intraspicific pair-matings. No significant correlations were found for D. melanogaster, but in D. simulans flicking by the female appeared to be associated with a shift in male wing display pattern resulting in higher levels of vibration. Flicking did not appear to discourage courtship by males in either species.

Analysis of courtship sequences of the hybrids between Drosophila melanogaster and Drosophila simulans.

Several transitions between sequential male courtship elements were analyzed for Drosophila melanogaster, its close relative D. simulans, and two types of hybrid males. Hybrid males from special reciprocal crosses did not differ. While D. melanogaster and D. simulans males differed markedly for the majority of transitions studied, hybrid males showed no consistent pattern with the parent species, being indistinguishable from D. simulans males, indistinguishable from D. melanogaster, or intermediate between them, depending on the trait observed. This suggests independent genetic control of these transitions during male courtship.

The morphology of the sex pheromone gland in Drosophila grimshawi.

The morphology of a sex pheromone-producing gland found in the abdomen of Drosophila grimshawi males was studied by light and electron microscopy. This gland, consisting of two intra-anal lobes, contains cells that resemble those of other insect pheromone glands. However, in contrast to many other insect pheromone glands that release pheromone through the cuticle, cells of the intra-anal lobes secrete into a canaliculi-duct system that empties into the anal region. The liquid secretory product flows along the surface of the intra-anal lobes and is brushed onto the substrate by fingerlike projections on the lobes' surfaces.