Effects of constant light on the rhythm of adult locomotor activity in the blowfly, Calliphora vicina

Abstract. Females of the blowfly, Calliphora vicina , showed an increase in the free-running period (γ) of their locomotor activity rhythm when transferred from continuous darkness (DD) to continuous 'dim' light (LL) at an irradiance below about 0.03 Wm^-2. Transfer to LL of this intensity also caused a reduction in the duration of the active phase (α) of the cycle. Transfer to 'bright' light (>0.03 Wm²), however, lead to arrhythmicity. Data suggest that constant light of this intensity does not 'stop' the circadian pacemaker but imposes behavioural arrhythmicity at a more superficial level.     

Effects of temperature and temperature-steps on circadian locomotor rhythmicity in the blow fly Calliphora vicina

The free-running period (in darkness) of the locomotor activity rhythm in adult blow flies (Calliphora vicina) was temperature-compensated between 15 and 25 degrees C, showing Q(10) values between 0.98 and 1.04. Single steps-up (20 to 25 degrees C) or steps-down (20 to 15 degrees C) in temperature caused stable phase shifts of the activity rhythm, giving rise to temperature-step phase response curves (PRCs) with both advances and delays. Phase advances, however, were dominant for steps-up, and phase delays for steps-down; the two PRCs were almost "mirror images" of each other. Following protocols introduced by Zimmerman et al. [(1968) Temperature compensation of the circadian oscillation in Drosophila pseudoobscura and its entrainment by temperature cycles, Journal of Insect Physiology, 14, 669-684] for the rhythm of pupal eclosion in Drosophila pseudoobscura, the steps-up and steps-down PRCs for C. vicina were used to compute a theoretical PRC for a 6 h low temperature pulse, and from this a theoretical steady-state phase relationship of the locomotor activity rhythm to a train of such pulses making up a temperature cycle (18 h at 20 degrees and 6 h at 15 degrees C).     

Under-sized larvae from short-day adults of the blow fly, Calliphora vicina, side-step the diapause programme

Abstract. Larvae of Calliphora vicina R.-D. (Diptera: Calliphoridae) hatching from eggs laid by adult females exposed to short day length, and then raised in darkness at 11^oC, normally enter diapause rather than undergoing prompt pupariation. However, if the feeding stage is curtailed by premature extraction of larvae from their food, or if the larvae are subjected to severe overcrowding, smaller larvae side-step the diapause programme to become miniature puparia, whereas larger larvae proceed to diapause as fully-fed ones. In addition, smaller diapausing larvae show a shorter (or less intense) diapause than full-sized larvae. Apart from the smallest individuals, flies can emerge from these miniature puparia and in some cases are capable of laying eggs; this suggests that avoidance of diapause may allow undersized individuals an opportunity to reproduce before winter sets in. Fat determinations performed on larvae and pupae of various sizes, however, showed that short-day (diapause-destined) individuals lay down the same proportion of fat as long-day (non-diapause) individuals, and the same proportion of fat across all size classes. The possible functional significance of this phenomenon is discussed.     

Maternal temperature has different effects on the photoperiodic response and duration of larval diapause in blow fly ( Calliphora vicina ) strains collected at two latitudes

Abstract .The blow fly Calliphora vicina Robineau-Desvoidy (Diptera: Calliphoridae) has a wide distribution across northern and temperate Europe. It has a facultative, maternally-induced larval diapause in response to short days. The photoperiodic response, measured at 15 and 20°C, of two populations was compared. A southern population (originating at 51° N) was sensitive to temperature at all daylengths; the incidence of diapause was greatly reduced at 20°C compared with 15°C. The photoperiodic response of a northern population (from 65° N) was sensitive to temperature only in long days; in short days (< 14 h of light) the response of this strain was identical at each temperature.
Variation in parental photoperiod and temperature were found to affect the duration of larval diapause, indicating a role for maternal effects in diapause intensity as well as incidence. However, the between-strain variation was greater than that within strains, indicating qualitative differences in diapause response. These differences may arise from the ecological conditions at the points of origin of the two strains. The northern strain from the harsher climate has a more intense diapause that follows a relatively temperature-insensitive photoperiodic response. In contrast, the southern strain has a shallow diapause and its photoperiodic response may be overridden by the experience of concurrent high temperature.     

Larval growth rates of the blowfly, Calliphora vicina, over a range of temperatures

Blowfly larvae (Diptera: Calliphoridae) fulfil an important ecological function in the decomposition of animal remains. They are also used extensively in forensic entomology, predominantly to establish a minimum time since death, or a minimum post-mortem interval, using the larval length as a 'biological clock'. This study examined the larval growth rate of a forensically important fly species, Calliphora vicina Robineau-Desvoidy (Diptera: Calliphoridae) at temperatures of between 4 degrees C and 30 degrees C, under controlled laboratory conditions. The laboratory flies had been trapped initially in London, U.K. The minimum developmental temperature was estimated to be 1 degrees C and 4700 accumulated degree hours (ADH) were required for development from egg hatch to the point of pupariation. Lines fitted to the laboratory larval growth data were found to adequately explain the growth of larvae in the field. The nature of variation in growth rates from geographically isolated populations is discussed.     

Inheritance of the photoperiodic response controlling larval diapause in the blow fly,Calliphora vicina

Larvae of the blow fly Calliphora vicina R-D. (Diptera: Calliphoridae) display a diapause in response to the exposure of their parents to short photoperiods. Due to geographic variation in photoperiodic response, flies from a southern, English population show a long-day response to the fixed photoperiod of L:D 15.5:8.5 whilst flies from a northern population from Finland show a short-day response to the same photoperiod. Crosses between these strains have shown previously that diapause incidence is a maternal characteristic; here we demonstrate that the hybrid female offspring of such crosses are not intermediate between the two parental strains but show a photoperiodic response biased towards their maternal line. Thus not only are males unable to influence directly the diapause incidence among their offspring but the indirect effects of inheritance down the male line are weaker than down the female. Diapause duration, in contrast, is influenced by each parent in a similar manner. Diapause lasts longer in larvae with a greater admixture of northern genes regardless of whether they were maternal or paternal.     

The effect of larval temperature and photoperiod on the incidence of larval diapause in the blowfly, Calliphora vicina

ABSTRACT. The blowfly, Calliphora vicina Meigen (Diptera: Calliphoridae), displays a maternally induced larval diapause. Progeny of adults exposed to short days enter diapause if the larval temperature is 15C or below; exposing adults to long days, or larvae to temperatures above 15C, results in non-diapause. By keeping progeny from short-day parents at the parental temperature of 23.5C for various lengths of time before transfer to 11C, it could be shown that the diapause-averting effect of high temperature operates in the late wandering stage of larva, possibly at the moment of the decision for the brain-ring gland complex to release PTTH/ecdysone at the diapause/non-diapause stage.
Although photoperiodic induction in the blowfly is maternal, previous work had shown that the larvae were also slightly sensitive to photoperiod. From the present study it is evident that also the eggs are photo-sensitive.     

Phase shifts of the adult locomotor activity rhythm in Calliphora vicina induced by non-steroidal ecdysteroid agonist RH 5849

RH 5849, a non-steroidal ecdysteroid mimic, was found to cause consistent phase shifts in the circadian rhythm of locomotor activity of the blowfly, Calliphora vicina. This compound causes phase advances in the early subjective night and phase delays in the late subjective night. This effect is the opposite, but not the mirror image of the phase response curve obtained for 1 h light pulses. This suggests that ecdysteroids might act as entraining agents via the output pathway by feedback to clock neurons in the brain. A computer model based on 12 pacemaker neurons with circadian periods ( values) from short to long without simulated feedback from the ecdysteroid system becomes arrhythmic; with feedback, the oscillators become synchronized to a common period. The possible role of ecdysteroids as endogenous synchronizing agents in the insect circadian system is discussed.     

Modulatory effects of octopamine and serotonin on male sensitivity and periodicity of response to sex pheromone in the cabbage looper moth,Trichoplusia ni

The pheromone-mediated flight behavior of male cabbage looper moths in a sustained-flight tunnel and random activity exhibited during scotophase were observed after males were treated with octopamine or serotonin (5 hydroxytryptamine). Octopamine induced a hypersensitivity to the olfactory signal, resulting in a significant lowering of the pheromone dose that elicited peak levels of response. Octopamine, however, did not affect the circadian rhythmicity of response to pheromone. In contrast, serotonin disrupted the circadian rhythmicity of response, resulting in a high percentage of males exhibiting random activity and response to pheromone throughout the entire 8 h scotophase instead of during the normal peak period during the latter part of the scotophase. Serotonin did not effect a decrease in the dose of pheromone eliciting peak response. In addition, at the highest dosages tested octopamine and serotonin induced opposite postures associated with a paralysis that occurred when males attempted to take flight to the pheromone.     

The role of the pineal and the retinae in the expression of circadian locomotor rhythmicity in the ruin lizard,Podarcis sicula

Summary A marked interspecific variability in the role played by the pineal and the retinae characterizes the circadian system of lizards. I examined the role played by these structures in a new model species, the ruin lizard, Podarcis sicula. In constant temperature and darkness pinealectomy as well as bilateral removal of the retinae produced significant changes (both lengthening and shortening) in the freerunning period of locomotor rhythms. Circadian activity time was also affected by pinealectomy. Circadian locomotor rhythmicity persisted in all cases even when both operations were combined in the same individuals. This demonstrates in Podarcis sicula the existence of an oscillatory system outside the pineal and the retinae which can drive locomotor rhythms. The period changes recorded after pinealectomy as well as after bilateral removal of the retinae specifically suggest that both the pineal and the retinae play a modulating role on circadian oscillators located elsewhere in the system, with the final effect of stabilizing the overt rhythms.Abbreviations DD constant darkness - LL constant light - PIN-X pinealectomy - RET-X bilateral removal of the retinae - SHAM sham pinealectomy - circadian activity time - freerunning circadian period     

Postembryonic development of serotonin-immunoreactive neurons in the central nervous system of the blowfly,Calliphora erythrocephala

Summary The postembryonic development of serotonin-immunoreactive (5-HTi) neurons was studied in the optic lobe of the blowfly. In the adult fly there are 24 5-HTi neurons invading each optic lobe. The perikarya of two of these neurons are situated in the dorso-caudal part of the protocerebrum (LBO-5HT neurons; large bilateral optic lobe 5-HTi neurons). The cell bodies of the remaining 22 neurons are located anteriorly at the medial base of the medulla (2 innervating the lobula, LO-5HT neurons; and 20 neurons innervating the medulla, ME-5HT neurons). The two central neurons (LBO-5HT neurons) are derived from metamorphosing larval neurons, while the ME- and LO-5HT neurons are imaginai optic lobe neurons differentiating during pupal development.The 5-HTi neurons of the optic lobe seem to have different ancestors. The LBO-5HT neurons are probably derived from segmental protocerebral neuroblasts, whereas the ME-and LO-5HT neurons are most likely derived from the inner optic anlage. The first 5-HTi fibers to reach the imaginal optic lobes are seen in the late third instar larva and are derived from the LBO-5HT neurons. The first ME- and LO-5HT neurons become immunoreactive at 24 h (10%) pupal development. At about 96 h (40%) of pupal development all the 5-HTi neurons of the optic lobes have differentiated and attained their basic adult morphology. The further development mainly entails increase in volume of arborizations and number of finer processes. The differentiation and outgrowth of 5-HTi processes follows that of, e.g., columnar neurons in the optic lobe neuropils. Hence, 5-HTi processes invade neuropil relatively late in the differentiation of the optic lobe.     

Ontogeny of the circadian system governing ecdysial rhythms in a holometabolous insect, Diatraea grandiosella (Pyralidae)

ABSTRACT. Three eclosion rhythms of Diatraea grandiosella Dyar (Lepidoptera, Pyralidae) showed different phase relationships to the LD cycle, different degrees of synchronization and different sensitivity to a brief light perturbation. Egg hatching occurred mainly at dawn, whereas pupation and adult emergence occurred mainly at dusk. The egg hatching was the most synchronized rhythm of the three, pupation the least. The egg hatching rhythm was more sensitive to a brief light perturbation than was the adult emergence rhythm. A light pulse in the middle of the night caused greater disturbance in the egg hatching rhythm than did a pulse placed in the early night. The three rhythms persisted in both DD and LL. LL free-running rhythm gradually became desynchronized and broke down at the third cycle. The phase relationships reveal difficulties in applying the 'external coincidence model' to this species' photoperiodic response.     

Presence of circadian rhythms in the locomotor activity of a cave-dwelling millipede Glyphiulus cavernicolus sulu (Cambalidae, Spirostreptida)

The locomotor activity of the millipede Glyphiulus cavernicolus (Spirostreptida), which occupies the deeper recesses of a cave, was monitored in light-dark (LD) cycles (12h light and 12h darkness), constant darkness (DD), and constant light (LL) conditions. These millipedes live inside the cave and are apparently never exposed to any periodic factors of the environment such as light-dark, temperature, and humidity cycles. The activity of a considerable fraction of these millipedes was found to show circadian rhythm, which entrained to a 12:12 LD cycle with maximum activity during the dark phase of the LD cycle. Under constant darkness (DD), 56.5% of the millipedes (n = 23) showed circadian rhythms, with average free-running period of 25.7h ± 3.3h (mean ± SD, range 22.3h to 35.0h). The remaining 43.5% of the millipedes, however, did not show any clear-cut rhythm. Under DD conditions following an exposure to LD cycles, 66.7% (n = 9) showed faint circadian rhythm, with average free-running period of 24.0h ± 0.8h (mean ± SD, range 22.9h to 25.2h). Under constant light (LL) conditions, only 2 millipedes of 11 showed free-running rhythms, with average period length of 33.3h ± 1.3h. The results suggest that these cave-dwelling millipedes still possess the capacity to measure time and respond to light and dark situations. (Chronobiology International, 17(6), 757-765, 2000)     

Involvement of cortisol and sirtuin1 during the response to stress of hypothalamic circadian system and food intake-related peptides in rainbow trout,Oncorhynchus mykiss

Stress is conditioning animal welfare by negatively affecting a wide range of physiological and behavioral functions. This may be applied to circadian physiology and food intake. Cortisol, the stress-related hormone, may mediate such effect of stress, but other indirect mediators might be considered, such as sirtuin1. Then, either the independent modulatory effect or the existence of any interaction between mediators may be responsible. The circadian system is the main modulator of several integrative mechanisms at both central and peripheral levels that are rhythmically presented, thus influencing different processes such as food intake. In this way, food intake is controlled by the circadian system, as demonstrated by the persistence of such rhythms of food intake in the absence of environmental external cues. Our study aimed to evaluate the daily profile of hypothalamic mRNA abundance of circadian clock genes (clock1a, bmal1, per1 and rev-erbβ-like), and food intake regulators (crf, pomc-a1, cart, and npy) in rainbow trout (Oncorhynchus mykiss), the impact of stress on such rhythms, and the involvement of cortisol and sirtuin1 as mediators. Four cohorts of trout were subjected to 1) normal stocking density (control group), 2) high stocking density for 72 hours (stress group), 3) normal stocking density and implanted with mifepristone, a glucocorticoid receptors antagonist, and 4) mifepristone administered and stressed for 72 hours. Fish from each group were sampled every 4-h along the 24-h LD cycle, and cortisol, glucose and lactate plasma levels were evaluated. Hypothalamic mRNA abundance of clock genes, food intake regulators, glucocorticoid receptors and sirtuin1 were qPCR assayed. Our results reveal the impact of stress on most of the genes assayed, but different mechanisms appear to be involved. The rhythm of clock genes displayed decreased amplitude and averaged levels in stressed trout, with no changes of the acrophase being observed. This effect was not prevented by mifepristone. On the contrary, the effect of stress on the daily profile of crf, pomc-a1, and npy was totally prevented by mifepristone administration. Accordingly, cortisol appears to mainly mediate the effect of stress on food intake regulators through binding to specific glucocorticoid receptors within trout hypothalamus, whereas sirtuin1 is apparently mediating such effects on the circadian system in the same brain region. Further research must be performed to clarify those mechanisms through which stress influences food intake and the circadian oscillator within the same brain region, hypothalamus, in rainbow trout, and the interaction among them all.     

Fluoxetine potentiates the effects of corticotropin-releasing factor on locomotor activity and serotonergic systems in the roughskin newt, Taricha granulosa

The anxiety- and stress-related neuropeptide corticotropin-releasing factor (CRF) elicits behavioral changes in vertebrates including increases in behavioral arousal and locomotor activity. Intracerebroventricular injections of CRF in an amphibian, the roughskin newt (Taricha granulosa), induces rapid increases in locomotor activity in both intact and hypophysectomized animals. We hypothesized that this CRF-induced increase in locomotor activity involves a central effect of CRF on serotonergic neurons, based on known stimulatory actions of serotonin (5-hydroxytryptamine, 5-HT) on spinal motor neurons and the central pattern generator for locomotor activity in vertebrates. In Experiment 1, we found that neither intracerebroventricular injections of low doses of CRF (25 ng) nor the selective serotonin reuptake inhibitor fluoxetine (10, 100 ng), by themselves, altered locomotor activity. In contrast, newts treated concurrently with CRF and fluoxetine responded with marked increases in locomotor activity. In Experiment 2, we found that increases in locomotor activity following co-administration of CRF (25 ng) and fluoxetine (100 ng) were associated with decreased 5-HT concentrations in a number of forebrain structures involved in regulation of emotional behavior and emotional states, including the ventral striatum, amygdala pars lateralis, and dorsal hypothalamus, measured 37 min after treatment. These results are consistent with the hypothesis that CRF stimulates locomotor activity through activation of serotonergic systems.     

Looking into the puparium: Micro‐CT visualization of the internal morphological changes during metamorphosis of the blow fly,Calliphora vicina,with the first quantitative analysis of organ development in cyclorrhaphous dipterans

Metamorphosis of cyclorrhaphous flies takes place inside a barrel‐like puparium, formed by the shrinking, hardening and darkening of the third‐instar larval cuticle. The opacity of this structure hampers the visualization of the morphological changes occurring inside and therefore a full understanding of the metamorphosis process. Here, we use micro‐computed tomography (micro‐CT) to describe the internal morphological changes that occur during metamorphosis of the blow fly, Calliphora vicina Robineau‐Desvoidy 1830 (Diptera: Calliphoridae) at a greater temporal resolution than anything hitherto published. The morphological changes were documented at 10% intervals of the total intra‐puparial period, and down to 2.5% intervals during the first 20% interval, when the most dramatic morphological changes occur. Moreover, the development of an internal gas bubble, which plays an essential role during early metamorphosis, was further investigated with X‐ray images and micro‐CT virtual sections. The origin of this gas bubble has been largely unknown, but micro‐CT virtual sections show that it is connected to one of the main tracheal trunks. Micro‐CT virtual sections also provided enough resolution for determining the completion of the larval‐pupal and pupal‐adult apolyses, thus enabling an accurate timing of the different intra‐puparial life stages. The prepupal, pupal, and pharate adult stages last for 7.5%, 22.5%, and 70% of the total intra‐puparial development, respectively. Furthermore, we provide for the first time quantitative data on the development of two organ systems of the blow fly: the alimentary canal and the indirect flight muscles. There is a significant and negative correlation between the volume of the indirect flight muscles and the pre‐helicoidal region of the midgut during metamorphosis. The latter occupies a large portion of the thorax during the pupal stage but narrows progressively as the indirect flight muscles increase in volume during the development of the pharate adult.     

Temporal flexibility in activity rhythms of a diurnal rodent,the ice rat (Otomys sloggetti)

ABSTRACT

Diurnality in rodents is relatively rare and occurs primarily in areas with low nighttime temperatures such as at high altitudes and desert areas. However, many factors can influence temporal activity rhythms of animals, both in the field and the laboratory. The temporal activity patterns of the diurnal ice rat were investigated in the laboratory with, and without, access to running wheels, and in constant conditions with running wheels. Ice rats appeared to be fundamentally diurnal but used their running wheels during the night. In constant conditions, general activity remained predominantly diurnal while wheel running was either nocturnal or diurnal. In some animals, entrainment of the wheel running rhythm was evident, as demonstrated by free-running periods that were different from 24 h. In other animals, the wheel running activity abruptly switched from nocturnal to subjective day as soon as the animals entered DD, and reverted back to nocturnal once returned to LD, suggesting the rhythms were masked by light. Wheel running rhythms appears to be less robust and more affected by light compared to general activity rhythms. In view of present and future environmental changes, the existence of more unstable activity rhythms that can readily switch between temporal niches might be crucial for the survival of the species.