Effects of photoperiod and aging on locomotor activity rhythms in the onion fly,Delia antiqua

At photoperiods longer than 8h per 24h, adults of the day-active onion fly Delia antiqua showed a major peak of locomotor activity in the late photophase and also bursts of activity induced by lights-on or lights-off. At shorter photoperiods the activity peaks fused. After transfer from long photoperiods to constant darkness (DD), the rhythm free-ran, but only the major peak persisted. This suggests that only the major peak is controlled by the circadian pacemaker. At long photoperiods, the daily phase of the major peak occurred progressively later with age. As a result, the activity at short photoperiods often shifted from photophase to scotophase in old flies. The free-running period (tau) also changed with age; tau was shorter than 24h until 14-20 days after eclosion and thereafter became longer, but a few individuals repeated changes in tau. The phase delay of locomotor activity with age in D. antiqua would be attributable to the increase in tau.     

Analysis of period mRNA cycling in Drosophila head and body tissues indicates that body oscillators behave differently from head oscillators.

The period (per) gene is thought to be part of the Drosophila circadian pacemaker. The circadian fluctuations in per RNA and protein that constitute the per feedback loop appear to be required for pacemaker function, and have been measured in head neuronal tissues that are necessary for locomotor activity and eclosion rhythms. The per gene is also expressed in a number of neuronal and nonneuronal body tissues for which no known circadian phenomena have been described. To determine whether per might affect some circadian function in these body tissues, per RNA cycling was examined. These studies show that per RNA cycles in the same phase and amplitude in head and body tissues during light-dark cycles. One exception to this is the lack of per RNA cycling in the ovary, which also appears to be the only tissue in which PER protein is primarily cytoplasmic. In constant darkness, however, the amplitude of per RNA cycling dampens much more quickly in bodies than in heads. Taken together, these results indicate that circadian oscillators are present in head and body tissues in which PER protein is nuclear and that these oscillators behave differently.     

A novel circadianly expressed Drosophila melanogaster gene dependent on the period gene for its rhythmic expression.

The Drosophila melanogaster period (per) gene is required for expression of endogenous circadian rhythms of locomotion and eclosion. per mRNA is expressed with a circadian rhythm that is dependent on Per protein; this feedback loop has been proposed to be essential to the central circadian pacemaker. This model would suggest the Per protein also controls the circadian expression of other genetic loci to generate circadian behavior and physiology. In this paper we describe Dreg-5, a gene whose mRNA is expressed in fly heads with a circadian rhythm nearly identical to that of the per gene. Dreg-5 mRNA continues to cycle in phase with that of per mRNA in conditions of total darkness and also when the daily feeding time is altered. Like per mRNA, Dreg-5 mRNA is not expressed rhythmically in per null mutant flies. Dreg-5 encodes a novel 298 residue protein and Dreg-5 protein isoforms also oscillate in abundance with a circadian rhythm. The phase of Dreg-5 protein oscillation, however, is different from that of Per protein expression, suggesting that Dreg-5 and per have common translational but different post-translational control mechanisms. These results demonstrate that the per gene is capable of modulating the rhythmic expression of other genes; this activity may form the basis of the output of circadian rhythmicity in Drosophila.     

The Drosophila dCREB2 gene affects the circadian clock

We report the role of dCREB2, the Drosophila homolog of CREB/CREM, in circadian rhythms. dCREB2 activity cycles with a 24 hr rhythm in flies, both in a light:dark cycle and in constant darkness. A mutation in dCREB2 shortens circadian locomotor rhythm in flies and dampens the oscillation of period, a known clock gene. Cycling dCREB2 activity is abolished in a period mutant, indicating that dCREB2 and Period affect each other and suggesting that the two genes participate in the same regulatory feedback loop. We propose that dCREB2 supports cycling of the Period/Timeless oscillator. These findings support CREB's role in mediating adaptive behavioral responses to a variey of environmental stimuli (stress, growth factors, drug addiction, circadian rhythms, and memory formation) in mammals and long-term memory formation and circadian rhythms in Drosophila.     

Circadian Locomotor Activity Under Artificial Light in the Freshwater Crab Pseudothelphusa americana

Long-term recordings of locomotor activity were obtained from intact freshwater crabs, Pseudothelphusa americana in constant darkness (DD), constant light (LL) and different light-dark (LD) protocols. Bimodal rhythms were typically observed in this crab when subjected to DD or LD, with bouts of activity anticipating lights-on and lights-off, respectively. Freerunning circadian rhythms were expressed in both DD and LL for longer than 30 days. In DD, we observed that some animals presented different period lengths for each activity component. During LL, activity was primarily unimodal, however spontaneous splitting of the rhythms were observed in some animals. When activity was recorded under artificial long days, the morning bouts maintained their phase relationship but the evening bouts changed their phase relationship with the Zeitgeber. Our results indicate that, bimodal locomotor activity rhythm in the crab Pseudothelphusa americana is variable among organisms. The characteristics of phase relationship with LD and responses to LL for morning and evening bouts, suggest that, locomotor activity could be driven by multiple oscillators, and that coupling between these oscillators may be regulated by light.     

Circadian Locomotor Activity Under Artificial Light in the Freshwater Crab Pseudothelphusa americana

Long-term recordings of locomotor activity were obtained from intact freshwater crabs, Pseudothelphusa americana in constant darkness (DD), constant light (LL) and different light-dark (LD) protocols. Bimodal rhythms were typically observed in this crab when subjected to DD or LD, with bouts of activity anticipating lights-on and lights-off, respectively. Freerunning circadian rhythms were expressed in both DD and LL for longer than 30 days. In DD, we observed that some animals presented different period lengths for each activity component. During LL, activity was primarily unimodal, however spontaneous splitting of the rhythms were observed in some animals. When activity was recorded under artificial long days, the morning bouts maintained their phase relationship but the evening bouts changed their phase relationship with the Zeitgeber. Our results indicate that, bimodal locomotor activity rhythm in the crab Pseudothelphusa americana is variable among organisms. The characteristics of phase relationship with LD and responses to LL for morning and evening bouts, suggest that, locomotor activity could be driven by multiple oscillators, and that coupling between these oscillators may be regulated by light.     

Circadian locomotor rhythms in individual honeybees

ABSTRACT. The circadian locomotor (walking) rhythms of individual forager honeybees ( Apis mellifera ligustica L.) were measured under a variety of conditions. In constant dark the rhythms exhibited endogenous periodicities that were less than 24 h, whereas under constant light the periods tended to be greater than 24 h. Individual honeybees readily entrained to photoperiods, displaying a diurnal pattern of entrainment with most of the activity occurring in late photophase. Evidence is presented which suggests that foraging behaviour and general locomotor behaviour may be governed by two different circadian clock systems.     

Pigment-dispersing factor affects nocturnal activity rhythms, photic entrainment, and the free-running period of the circadian clock in the cricket gryllus bimaculatus

Pigment-dispersing factor (PDF) is a neuropeptide widely distributed in insect brains and plays important roles in the circadian system. In this study, we used RNA interference to study the role of the pigment-dispersing factor (pdf) gene in regulating circadian locomotor rhythms in the cricket, Gryllus bimaculatus. Injections of pdf double-stranded RNA (dspdf) effectively knocked down the pdf mRNA and PDF peptide levels. The treated crickets maintained the rhythm both under light-dark cycles (LD) and constant darkness (DD). However, they showed rhythms with reduced nocturnal activity with prominent peaks at lights-on and lights-off. Entrainability of dspdf-injected crickets was higher than control crickets as they required fewer cycles to resynchronize to the LD cycles shifted by 6 h. The free-running periods of the dspdf-injected crickets were shorter than those of control crickets in DD. These results suggest that PDF is not essential for the rhythm generation but involved in control of the nocturnality, photic entrainment, and fine tuning of the free-running period of the circadian clock.     

Novel wheel running blocks the preovulatory luteinizing hormone surge and advances the hamster circadian pacemaker

In rodents, the preovulatory luteinizing hormone (LH) surge is timed by a circadian rhythm. We recently reported that a phenobarbital-induced delay of the estrous cycle in Syrian hamsters is associated with an approximately 2-h phase advance in both the circadian locomotor activity rhythm and the timing of the LH surge. The following study tests the hypothesis that a >2-h nonpharmacological phase advance in the circadian pacemaker that delays the estrous cycle by a day will also phase advance the LH surge by approximately 2 h. Activity rhythms were continuously monitored in regularly cycling hamsters using running wheels or infrared detectors for about 10 days prior to jugular cannulation. The next day, on proestrus, hamsters were transferred to the laboratory for 1 of 3 treatments: transfer to a "new cage" (and wheel) from zeitgeber time (ZT) 4 to 8 (with ZT12 defined as time of lights-off), or exposure to a "novel wheel" at ZT5 or ZT1. All animals were then placed in constant dark (DD). Blood samples were obtained just before onset of DD and hourly for the next 6 h, on that day and the next day for determination of plasma LH concentrations. Running activity was monitored in DD for about 10 more days. Transfer to a novel wheel at either ZT5 or ZT1 delayed the LH surge to day 2 in most hamsters, whereas exposure to a new cage did not. Only the delayed LH surges were phase advanced at least 2.5 h on average in all 3 groups. However, wheel-running activity was similarly phase advanced in all 3 groups regardless of the timing of the LH surge; thus, the phase advances in circadian activity rhythms were not associated with the 1-day delay of the LH surge. Interestingly, the number of wheel revolutions was closely associated with the 1-day delay of LH surges following exposure to a novel wheel at either ZT1 or ZT5. These results suggest that the intensity of wheel running (or an associated stimulus) plays an important role in the circadian timing mechanism for the LH surge.     

Photoperiodic time measurement in the male deer mouse, Peromyscus maniculatus

Weanling male deer mice, Peromyscus maniculatus, were exposed for three weeks either to light-dark (LD) cycles with periods (T=L+D) ranging from T=23 (1L:22D) to T=25.16 (1L:24.16D) or to 24-h LD cycles with photoperiods ranging from 1 (1L:23D) to 19 (19L:5D) h. Both the circadian locomotor activity rhythms and the response of the reproductive system to these LD cycles were assessed. The results demonstrate that the photoperiodic effectiveness of light depends on the phase of the light relative to the animal's circadian system, as marked by the circadian activity rhythm. Light falling during the animal's subjective night, from activity onset to at least 11.8 h after activity onset, stimulates growth and maturation of the reproductive system, whereas light falling during the rest of the circadian cycle is nonstimulatory.     

Dopaminergic Regulation of Circadian Food Anticipatory Activity Rhythms in the Rat

Circadian activity rhythms are jointly controlled by a master pacemaker in the hypothalamic suprachiasmatic nuclei (SCN) and by food-entrainable circadian oscillators (FEOs) located elsewhere. The SCN mediates synchrony to daily light-dark cycles, whereas FEOs generate activity rhythms synchronized with regular daily mealtimes. The location of FEOs generating food anticipation rhythms, and the pathways that entrain these FEOs, remain to be clarified. To gain insight into entrainment pathways, we developed a protocol for measuring phase shifts of anticipatory activity rhythms in response to pharmacological probes. We used this protocol to examine a role for dopamine signaling in the timing of circadian food anticipation. To generate a stable food anticipation rhythm, rats were fed 3h/day beginning 6-h after lights-on or in constant light for at least 3 weeks. Rats then received the D2 agonist quinpirole (1 mg/kg IP) alone or after pretreatment with the dopamine synthesis inhibitor α-methylparatyrosine (AMPT). By comparison with vehicle injections, quinpirole administered 1-h before lights-off (19h before mealtime) induced a phase delay of activity onset prior to the next meal. Delay shifts were larger in rats pretreated with AMPT, and smaller following quinpirole administered 4-h after lights-on. A significant shift was not observed in response to the D1 agonist SKF81297. These results provide evidence that signaling at D2 receptors is involved in phase control of FEOs responsible for circadian food anticipatory rhythms in rats.     

Possible evidence for morning and evening oscillators in Drosophila melanogaster populations selected for early and late adult emergence

In this paper, we report the results of our study aimed at a systematic analysis of the circadian phenotypes of fruit flies Drosophila melanogaster selected for early and late adult emergence, in light of the "morning and evening oscillator" (M and E) model for circadian clocks. We monitored adult emergence and activity/rest rhythms in these flies under light/dark (LD) cycles with short (8:16 h), normal (12:12 h) and long (16:8 h) photoperiods, as well as under constant darkness (DD). Across all the three LD cycles, the early populations displayed a morning phenotype with peak of emergence and activity occurring earlier than the controls and greater anticipation to "lights-on" and weak anticipation to "lights-off", while the late populations showed an evening phenotype with peak of emergence and activity occurring later than the controls and greater anticipation to lights-off and weak anticipation to lights-on. The gate of adult emergence and duration of activity in the early populations was narrower than the controls, while those of the late populations were wider than the controls. In addition, the circadian periodicities of adult emergence and activity/rest rhythms of the early flies were significantly shorter than the controls, while those of the late flies were significantly longer than the controls. In summary, the circadian phenotypes indicate that the early populations have evolved a dominant M oscillator, while the late populations have evolved a dominant E oscillator, thus providing an empirical support for the M and E model in Drosophila.