Entrainment of eclosion rhythm in Drosophila melanogaster populations reared for more than 700 generations in constant light environment

In this paper, we report the results of our extensive study on eclosion rhythm of four independent populations of Drosophila melanogaster that were reared in constant light (LL) environment of the laboratory for more than 700 generations. The eclosion rhythm of these flies was assayed under LL, constant darkness (DD) and three periodic light-dark (LD) cycles (T20, T24, and T28). The percentage of vials from each population that exhibited circadian rhythm of eclosion in DD and in LL (intensity of approximately 100 lux) was about 90% and 18%, respectively. The mean free-running period (τ) of eclosion rhythm in DD was 22.85 ± 0.87 h (mean ± SD). Eclosion rhythm of these flies entrained to all the three periodic LD cycles, and the phase relationship (ψ) of the peak of eclosion with respect to “lights-on” of the LD cycle was significantly different in the three periodic light regimes (T20, T24, and T28). The results thus clearly demonstrate that these flies have preserved the ability to exhibit circadian rhythm of eclosion and the ability to entrain to a wide range of periodic LD cycles even after being in an aperiodic environment for several hundred generations. This suggests that circadian clocks may have intrinsic adaptive value accrued perhaps from coordinating internal metabolic cycles in constant conditions, and that the entrainment mechanisms of circadian clocks are possibly an integral part of the clockwork.     

Genetic architecture underlying morning and evening circadian phenotypes in fruit flies Drosophila melanogaster

Circadian rhythms are perhaps among the genetically best characterized behaviours. Several mutations with drastic effects on circadian processes have been identified and models developed to explain how clock genes and their products generate self-sustained oscillations. Although natural variations in circadian phenotypes have been studied extensively, the genetic basis of such adaptive variations remains largely unknown. Here we report the results of a preliminary genetic analysis of adaptive divergence of circadian phenotypes in populations of fruit flies Drosophila melanogaster. Two sets of populations, ‘early'' and ‘late'', were created in a long-term laboratory selection for morning and evening emergence, with four independent replicates each. Over the course of ∼55 generations, the early flies evolved increased morning emergence and a shorter circadian period, whereas late flies evolved increased evening emergence and longer period. To examine the genetic basis of circadian phenotypes, we set up crosses between early and late flies, and monitored emergence and activity/rest rhythms in the F1, backcrossed and F2 progeny. Our analysis suggests that the genetic basis of divergent circadian phenotypes in early and late stocks is primarily autosomal. Line-cross analysis revealed that additive and non-additive genetic effects contribute to the divergence of circadian phenotypes in early and late flies.     

The circadian system of Drosophila melanogaster and its light input pathways

The fruit fly Drosophila melanogaster has been a grateful object for circadian rhythm researchers over several decades. Behavioral, genetic, and molecular studies in the little fly have aided in understanding the bases of circadian time keeping and rhythmic behaviors not only in Drosophila, but also in other organisms, including mammals. This review summarizes our present knowledge about the fruit fly's circadian system at the molecular and neurobiological level, with special emphasis on its entrainment by environmental light-dark cycles. The results obtained for Drosophila are discussed with respect to parallel findings in mammals.     

Latitude dependent arrhythmicity in the circadian oviposition rhythm of Drosophila ananassae

Latitude dependent arrhythmicity in the circadian rhythm of oviposition of Drosophila ananassae strains originating from 8.1°N to 32.7°N was studied by inbreeding them in cycles of 12 h of light at 20 lux and 12 h of darkness. The number of inbreeding generations required to initiate arrhythmicity in oviposition rhythm was dependent on the origin of latitude of the strain. The strains from the lower latitudes became arrhythmic after notably more numbers of generations than those from the higher latitudes. This might be attributed to the higher inherent degree of oviposition rhythmicity in the F₁ generation, and enhanced photic sensitivity of the circadian pacemaker mediating entrainment of oviposition rhythm of the strains from lower latitudes as compared to those from the higher latitudes.     

Behavioural circadian rhythms measured in real-time by automatic image analysis: applications in parasitoid insects

Abstract. A video analysis system was developed to study behavioural circadian rhythms in insects. Technical innovations were performed in image analysis and in the use of robotics. In image analysis, both the automatic detection of the insect and the calculation of several variables (linear and angular speed, locomotor activity, etc.) are carried out in real-time, which saves time and allows long-term studies without any intervention of the operator. The displacement of the camera by robotics allows the simultaneous study of many individuals (i.e. sixty individuals for the study of locomotor activity of adult insects, more than 400 individuals for the study of the emergence of adult parasitoids from their host). Four applications of this new device are presented. They deal with circadian rhythms in parasitoid insects: locomotor activity and time variation in locomotory parameters, daily distribution of the emergence and onset of the locomotor activity in adults. The adaptability of the system to several experimental materials and situations is discussed and the interest of the study of behavioural rhythms in the approach of host-parasitoid associations is emphasized.     

Rhythmic egg-laying behaviour in virgin females of fruit flies Drosophila melanogaster

Fruit fly Drosophila melanogaster females display rhythmic egg-laying under 12:12?h light/dark (LD) cycles which persists with near 24?h periodicity under constant darkness (DD). We have shown previously that persistence of this rhythm does not require the neurons expressing pigment dispersing factor (PDF), thought to be the canonical circadian pacemakers, and proposed that it could be controlled by peripheral clocks or regulated/triggered by the act of mating. We assayed egg-laying behaviour of wild-type Canton S (CS) females under LD, DD and constant light (LL) conditions in three different physiological states; as virgins, as females allowed to mate with males for 1?day and as females allowed to mate for the entire duration of the assay. Here, we report the presence of a circadian rhythm in egg-laying in virgin D. melanogaster females. We also found that egg-laying behaviour of 70 and 90% females from all the three male presence/absence protocols follows circadian rhythmicity under DD and LL, with periods ranging between 18 and 30?h. The egg-laying rhythm of all virgin females synchronized to LD cycles with a peak occurring soon after lights-off. The rhythm in virgins was remarkably robust with maximum number of eggs deposited immediately after lights-off in contrast to mated females which show higher egg-laying during the day. These results suggest that the egg-laying rhythm of D. melanogaster is endogenously driven and is neither regulated nor triggered by the act of mating; instead, the presence of males results in reduction in entrainment to LD cycles.     

Daily rhythms in the sorghum shootfly, Atherigona soccata: oviposition, egg-hatch and adult eclosion

ABSTRACT. Daily rhythms controlling oviposition, egg-hatching and adult eclosion in the sorghum shootfly, Atherigona soccata Rondani, were investigated. Eggs were laid only during the photophase of a LD 12:12 cycle, in two peaks. Under continuous light, this oviposition was considerably attenuated but not made immediately arrhythmic. Egg-hatching and adult eclosion both commenced just before dawn. Some feature of the scotophase during or immediately after black-head formation apparently acts as a signal for hatching. Eclosion was controlled by light but its timing in the field was modified by temperature. The last 2–3 days of the pupal period constituted the most sensitive stage, and light signals received during this period determined the time of eclosion. Ecological advantages of these rhythms to the shootfly are discussed.     

Extent of mismatch between the period of circadian clocks and light/dark cycles determines time‐to‐emergence in fruit flies

Circadian clocks time developmental stages of fruit flies Drosophila melanogaster, while light/dark (LD) cycles delimit emergence of adults, conceding only during the “allowed gate.” Previous studies have revealed that time‐to‐emergence can be altered by mutations in the core clock gene period (per), or by altering the length of LD cycles. Since this evidence came from studies on genetically manipulated flies, or on flies maintained under LD cycles with limited range of periods, inferences that can be drawn are limited. Moreover, the extent of shortening or lengthening of time‐to‐emergence remains yet unknown. In order to pursue this further, we assayed time‐to‐emergence of D. melanogaster under 12 different LD cycles as well as in constant light (LL) and constant dark conditions (DD). Time‐to‐emergence in flies occurred earlier under LL than in LD cycles and DD. Among the LD cycles, time‐to‐emergence occurred earlier under T4–T8, followed by T36–T48, and then T12–T32, suggesting that egg‐to‐emergence duration in flies becomes shorter when the length of LD cycles deviates from 24 h, bearing a strong positive and a marginally negative correlation with day length, for values shorter and longer than 24 h, respectively. These results suggest that the extent of mismatch between the period of circadian clocks and environmental cycles determines the time‐to‐emergence in Drosophila.     

Suppression of circadian rhythms of pineal and testicular hormones following lithium treatment in normal and reversed light-dark cycles, constant light and constant dark in rats

The aim of the current investigation was to study the effect of lithium on circadian rhythms of pineal - testicular hormones by quantitations of pineal and serum serotonin, N-acetylserotonin and melatonin, and serum testosterone at four time points (06.00, 12.00, 18.00 and 24.00) of a 24-hr period under normal photoperiod (L:D), reversed photoperiod (D:L), constant light (L:L) and constant dark phase (D:D) in rats. Circadian rhythms were observed in pineal hormones in all the combinations of photoperiodic regimens, except in constant light, and in testosterone levels in all the photoperiodic combinations. Pineal and serum N-acetylserotonin and melatonin levels were higher than serotonin at night (24.00 hr), in natural L:D cycle, in reversed L:D cycle or similar to normal L:D cycle in constant dark phase, without any change in constant light. In contrast, testosterone level was higher in light phase (12.00 hr through 18.00 hr) than in the dark phase (24.00 hr through 06.00 hr) in normal L:D cycle, in reversed L:D cycle, similar to normal L:D cycle in constant dark (D:D), and reversed to that of the normal L:D cycle in constant light (L:L). Lithium treatment (2 mEq/kg body weight daily for 15 days) suppressed the magnitude of circadian rhythms of pineal and serum serotonin, N-acetylserotonin and melatonin, and testosterone levels by decreasing their levels at four time points of a 24-hr period in natural L:D or reversed D:L cycle and in constant dark (D:D). Pineal indoleamine levels were reduced after lithium treatment even in constant light (L:L). Moreover, lithium abolished the melatonin rhythms in rats exposed to normal (L:D) and reversed L:D (D:L) cycles, and sustained the rhythms in constant dark. But testosterone rhythm was abolished after lithium treatment in normal (L:D)/reversed L:D (D:L) cycle or even in constant light/dark. The findings indicate that the circadian rhythm exists in pineal hormones in alternate light - dark cycle (L:D/D:L) and in constant dark (D:D), but was absent in constant light phase (L:L) in rats. Lithium not only suppresses the circadian rhythms of pineal hormones, but abolishes the pineal melatonin rhythm only in alternate light - dark cycles, but sustains it in constant dark. The testosterone rhythm is abolished after lithium treatment in alternate light - dark cycle and constant light/dark. It is suggested that (a) normal circadian rhythms of pineal hormones are regulated by pulse dark phase in normal rats, (b) lithium abolishes pineal hormonal rhythm only in pulse light but sustains it in constant dark phase, and (c) circadian testosterone rhythm occurs in both pulse light or pulse dark phase in normal rats, and lithium abolishes the rhythm in all the combinations of the photoperiod. The differential responses of circadian rhythms of pineal and testicular hormones to pulse light or pulse dark in normal and lithium recipients are discussed.     

Maternity-related plasticity in circadian rhythms of bumble-bee queens

Unlike most animals studied so far in which the activity with no circadian rhythms is pathological or linked to deteriorating performance, worker bees and ants naturally care for their sibling brood around the clock with no apparent ill effects. Here, we tested whether bumble-bee queens that care alone for their first batch of offspring are also capable of a similar chronobiological plasticity. We monitored locomotor activity of Bombus terrestris queens at various life cycle stages, and queens for which we manipulated the presence of brood or removed the ovaries. We found that gynes typically emerged from the pupae with no circadian rhythms, but after several days showed robust rhythms that were not affected by mating or diapauses. Colony-founding queens with brood showed attenuated circadian rhythms, irrespective of the presence of ovaries. By contrast, queens that lost their brood switched again to activity with strong circadian rhythms. The discovery that circadian rhythms in bumble-bee queens are regulated by the life cycle and the presence of brood suggests that plasticity in the circadian clock of bees is ancient and related to maternal behaviour or physiology, and is not a derived trait that evolved with the evolution of the worker caste.     

Experimental evolution in Drosophila melanogaster: interaction of temperature and food quality selection regimes

Abstract. In Drosophila , both the phenotypic and evolutionary effect of temperature on adult size involves alterations to larval resource processing and affects other life-history traits, that is, development time but most notably, larval survival. Therefore, thermal evolution of adult body size might not be independent of simultaneous adaptation of larval traits to resource availability. Using experimental evolution lines adapted to high and low temperatures at different levels of food, we show that selection pressures interact in shaping larval resource processing. Evolution on poor food invariably leads to lower resource acquisition suggesting a cost to feeding behavior. However, following low temperature selection, lower resource acquisition led to a higher adult body size, probably by more efficient allocation to growth. In contrast, following high temperature selection, low resource acquisition benefited larval survival, possibly by reducing feeding-associated costs. We show that evolved differences to larval resource processing provide a possible proximate mechanism to variation in a suite of correlated life-history traits during adaptation to different climates. The implication for natural populations is that in nature, thermal evolution drives populations to opposite ends of an adult size versus larval survival trade-off by altering resource processing, if resource availability is limited.     

黑腹果蝇种组的核型与进化研究

观察了国内黑腹果蝇种组34种果蝇的有丝分裂中期核型,其中首次描述了一些新核型。系统地分析了黑腹果蝇种组8个种亚组之间的核型进化关系及种间亲缘关系。结果是:elegans种亚组的核型为A型;eugracilis、melanogaster和ficusphila种亚组的核型为C型;takahashii和suzukii种亚组的核型为C型和D型;montium种亚组的核型为B、C、C’、D、D’、和E型;ananassae种亚组的核型为F、G和H型。从核型分化的角度可以将黑腹果蝇种组分为5个谱系:elegans,eugracilis-melanogaster-ficusphila,takkahashii-suzukii,montium,ananassae。这与2004年Yang等的观点基本一致,正好从核型进化的角度验证了Yang通过DNA序列分析所得到的结果。差别只在于elegans种亚组,作者把它单独列为一支,认为是祖先种亚组。通过选取同一种果蝇的几个不同地域单雌系的核型分析,结果表明:同一种果蝇的核型存在地域差异。这种差异可能是由于不同生境造成,也可能是本身进化程度的差异,或是两种因素相互作用的结果。     

Circadian rhythms and the evolution of photoperiodic timing in insects

This review discusses possible evolutionary trends in insect photoperiodism, mainly from a chronobiological perspective. A crucial step was the forging of a link between the hormones regulating diapause and the systems of biological rhythms, circadian or circannual, which have independently evolved in eukaryotes to synchronize physiology and behaviour to the daily cycles of light and darkness. In many of these responses a central feature is that the circadian system resets to a constant phase at the beginning of the subjective night, and then ‘measures’ the duration of the next scotophase. In ‘external coincidence’, one version of such a clock, light now has a dual role. First, it serves to entrain the circadian system to the stream of pulses making up the light/dark cycle and, second, it regulates the nondiapause/diapause switch in development by illuminating/not illuminating a specific light sensitive phase falling at the end of the critical night length. Important work by A. D. Lees on the aphid Megoura viciae using so‐called ‘night interruption experiments' demonstrates that pulses falling early in the night lead to long‐day effects that are reversible by a subsequent dark period longer than the critical night length and also show maximal sensitivity in the blue–green range of the spectrum. Pulses falling in the latter half of the night, however, produce long‐day effects that are irreversible by a subsequent long‐night and show a spectral sensitivity extending into the red. With movement to higher latitudes, insects develop genetic clines in various parameters, including critical night length, the number of long‐night cycles needed for diapause induction, the strength of the response, and the ‘depth’ or intensity of the diapause thus induced. Evidence for these and other types of photoperiodic response suggests that they provided strong selective advantages for insect survival.     

Plasticity of diel and circadian activity rhythms in fishes

In many fish species, some individuals arediurnal while others are nocturnal. Sometimes,the same individual can be diurnal at first andthen switch to nocturnalism, or vice-versa.This review examines the factors that areassociated with such plasticity. It covers thebreakdown of activity rhythms during migration,spawning, and the parental phase; reversals ofactivity patterns during ontogeny or from oneseason to the next; effects of light intensity,temperature, predation risk, shoal size, foodavailability, and intraspecific competition.Case studies featuring goldfish (Carassiusauratus), golden shiner (Notemigonuscrysoleucas), lake chub (Couesiusplumbeus), salmonids, sea bass (Dicentrarchus labrax), and parentalsticklebacks and cichlids illustrate some ofthese influences. It is argued that mostspecies have a circadian system but that havingsuch a system does not necessarily imply strictdiurnalism or nocturnalism. Rigidity ofactivity phase seems more common in species,mostly marine, that display behavioral sleep,and for these animals the circadian clock canhelp maintain the integrity of the sleepperiod and ensure that its occurrence takes place atthat time of day to which the animal's sensoryequipment is not as well adapted. However, inother fishes, mostly from freshwater habitats,the circadian clock seems to be used mainly foranticipation of daily events such as thearrival of day, night, or food, and possiblyfor other abilities such as time-place learningand sun compass orientation, rather than forstrict control of activity phase. In thesespecies, various considerations relating toforaging success and predation risk maydetermine whether the animal is diurnal ornocturnal at any particular time and place.     

On-line data acquisition and evaluation of long-term measurements of circadian rhythms in individual cells ofAcetabularia

A multitasking time-sharing computer system was implemented for studies of different circadian rhythms in individual cells of the unicellular green alga,Acetabularia. This fully automatized system allows simultaneous data acquisition and analysis. Graphical presentation of untreated and mathematically treated data is permanently available on three graphic displays and on a digital plotter. The sampling rate for the data acquisition in each of the 60 channels connected to the system is 720/24 h. Provisions have been made to guarantee uninterrupted data uptake for these long-term measurements by including an auto-restart module and by providing extremely reliable software for the experimenter using menu techniques.     

Circadian clock properties of fruit flies Drosophila melanogaster exhibiting early and late emergence chronotypes

The role of circadian clocks in timing daily behaviors is widely acknowledged, and while empirical evidence suggests that clock period is correlated with the preferred phase of a rhythmic behavior (chronotype), other clock properties have also been hypothesized to underlie chronotype variation. Here, we report that fruit fly Drosophila melanogaster populations exhibiting evening emergence chronotype (late) are characterized by higher incidence of behavioral arrhythmicity in constant dim light, wider range of entrainment, reduced rates of re-entrainment to simulated jet-lag and higher amplitude of both entrained and free-running rhythms as compared to those exhibiting morning emergence chronotype (early). Our results thus highlight the role of circadian clock properties such as zeitgeber sensitivity, amplitude and coupling in driving chronotype variation.     

Evolution of increased adult longevity in Drosophila melanogaster populations selected for adaptation to larval crowding

In holometabolous animals such as Drosophila melanogaster, larval crowding can affect a wide range of larval and adult traits. Adults emerging from high larval density cultures have smaller body size and increased mean life span compared to flies emerging from low larval density cultures. Therefore, adaptation to larval crowding could potentially affect adult longevity as a correlated response. We addressed this issue by studying a set of large, outbred populations of D. melanogaster, experimentally evolved for adaptation to larval crowding for 83 generations. We assayed longevity of adult flies from both selected (MCUs) and control populations (MBs) after growing them at different larval densities. We found that MCUs have evolved increased mean longevity compared to MBs at all larval densities. The interaction between selection regime and larval density was not significant, indicating that the density dependence of mean longevity had not evolved in the MCU populations. The increase in longevity in MCUs can be partially attributed to their lower rates of ageing. It is also noteworthy that reaction norm of dry body weight, a trait probably under direct selection in our populations, has indeed evolved in MCU populations. To the best of our knowledge, this is the first report of the evolution of adult longevity as a correlated response of adaptation to larval crowding.     

Circadian rhythms of biomarkers of oxidative stress and their characteristics in broiler chickens reared under natural light/dark cycle

For the purpose of studying circadian rhythms of biomarkers of oxidative stress in broiler chickens maintained under natural photoperiod, we collected blood from the wing veins of 10 matured broiler chickens every three hours for a period of 24 h. The blood samples were analyzed for superoxide dismutase (SOD), catalase (CAT), and malondialdehyde (MDA). The fitting in of single cosinor procedure showed that CAT and MDA exhibited clear circadian rhythms, while SOD did not. The amplitude of daily rhythms for the parameters was small, while the acrophases were all restricted to the light phase of the light:dark cycle. In conclusion, to our knowledge, this is the first time investigation of the circadian rhythms of antioxidants in broilers chickens, reared under natural photoperiod in the tropics. The result may be exploited for a more precise targeting of reactive oxygen species, thereby offering better protection for the cells to combat oxidative stress.