Possible involvement of nitric oxide in generation of orcadian rhythm

Abstract

The effect of continuous infusion of N^G‐methylarginine, an inhibitor of nitric oxide synthase, into the third cerebral ventricle above the suprachiasmatic nucleus (SCN) of the hypothalamus on the circadian rhythm of water intake was examined in rats maintained under either a 12‐h light and 12‐h dark cycle or in constant darkness. N^G‐methylarginine disrupted the circadian rhythm in both conditions. In constant darkness, however, the effect of the inhibitor on the rhythm was found to be due to change in its phase. These findings suggest that nitric oxide is involved in the mechanism of the generation and/or synchronization of the circadian rhythm driven by the circadian oscillator in the SCN.     

CIRCADIAN RHYTHM IN GROWTH AND DEATH OF ANABAENA FLOS-AQUAE (CYANOBACTERIA)

Circadian periodicity in cell division and death was investigated in the cyanobacterium Anabaena flos-aquae (Lyngb.) Bréb in a phosphorus (P)-limited, N₂-fixing chemostat culture. When entrained under 12:12 h LD cycles, not only cell division but also cell death showed a clear circadian rhythm in this filamentous cyanobacterium. The rhythm persisted under continuous light and was temperature compensated. Circadian rhythm was clearly observed in the steady-state cell number and instantaneous growth rate, μ(t), which reached a maximum at about 2 h before sunset and a minimum at about 2 h before sunrise. The number of dead cells and the instantaneous death rate γ(t) also showed a circadian periodicity; the peak of γ(t) occurred approximately 8 h before that of μ(t). Therefore, cell growth and death in A. flos-aquae appear to be under the control of circadian clocks, and thus it seems that their death is programmed cell death.     

About the Existence of Circadian Activity in Cave Crayfish

Evidence of a circadian clock mechanism was found in the cave crayfish Procambarus cavernicola. Analysis of motor activity recorded in this species during 12 consecutive days in either free running (constant darkness, DD or constant light, LL) or entrainment conditions (12 h of light alternated with 12 h of darkness, 12 : 12 LD) showed a well recognized circadian rhythm. In this rhythm however, the absence of synchronization by periodical external signals was notorious. The comparison between the motor circadian rhythm in cave crayfish and epigeous crayfish Procambarus clarkii (these last studied during juvenile and adult stages), evidenced strong similitude between the motor circadian rhythm of cave crayfish and juvenile epigeous crayfish.     

Conditionality of circadian rhythmicity: Synergistic action of light and temperature

Summary With cells which have been grown at 20°C, the circadian rhythm of bioluminescence inGonyaulax polyedra disappears at a critical temperature, which is about 12°C. The transition from the rhythmic to the arrhythmic state is very sharp with temperature: the two states are separated by only 1–2°C. Following a return to a higher temperature (20°C) under otherwise constant conditions, the rhythm resumes with its new phase defined by the time of the cool to warm transition. Loss of rhythmicity also occurs in constant bright light, with a similar resumption and phase determination upon transfer to darkness. The experiments described here show that the effects of light and low temperature are additive: rhythmicity is lost under combined low temperature and light intensity treatments which are ineffective individually.Abbreviations CT circadian time - ft-c footcandle - LD 12:12 12 h light/12 h dark cycle NIH Predoctoral Trainee in Biophysics, 2 T01 GM00782-16.     

Human cone light sensitivity and melatonin rhythms following 24-hour continuous illumination

This study investigates the possibility of an endogenous circadian rhythm in retinal cone function in humans. A full-field cone electroretinogram (ERG) was performed every 2 h for 24 h under continuous rod-saturating ambient white light (53 ± 30 lux; pupils dilated) in nine healthy subjects. Distinct circadian variations were superimposed upon a gradual decrease in cone responsiveness to light, demonstrated most reliably in the implicit times of b-wave and oscillatory potentials, and to a lesser extent in amplitude and a-wave implicit times. After mathematical correction of the linear trend, the cone response was found to be greatest around 20:00 h and least around 06:00 h. The phase of the ERG circadian rhythm was not synchronized with the phase of the salivary melatonin rhythm measured the previous evening. Melatonin levels measured under constant light on the day of ERG assessments were suppressed by 53% on average compared to melatonin profiles obtained previously under near-total darkness in seven participants. The progressive decline in cone responsiveness to light over the 24 h may reflect an adaptation of the cone-driven retinal system to constant light, although the mechanism is unclear. The endogenous rhythm of cone responsiveness to light may be used as an additional index of central or retinal circadian clock time.     

Circadian rhythm of a TCA cycle enzyme is apparently regulated at the translational level in the dinoflagellate Lingulodinium polyedrum

Previously, the authors have reported that intracellular amounts of several metabolic-related enzymes from the photosynthetic dinoflagellate Lingulodinium polyedrum(formerly Gonyaulax polyedra) showed a daily rhythm under a 12:12 h LD cycle. This led the authors to hypothesize that a circadian clock controls metabolism, including the tricarboxylic acid (TCA) cycle. In this study, the authors investigated daily changes in the levels of mRNA, protein, and enzyme activity of several metabolic enzymes during 12:12 h LD, 8:16 h LD, and constant light conditions. The NADP-dependent isocitrate dehydrogenase (NADPICDH) in the TCA cycle exhibited circadian changes of protein abundance and enzyme activity under all conditions, whereas its mRNA level remained constant throughout the cycle. These results indicate that the rhythm of NADPICDH is regulated by a circadian control of protein synthesis or modification rather than by message levels and suggest that the TCA cycle may be controlled by the circadian clock system.     

Effect of Light Intensity on Circadian Activity in Developing Japanese Quail

The effects of constant light on the expression of the circadian rhythm of feeding activity in Japanese quail, and in particular on the clarity of the rhythm were investigated. We used 46 4-week-old birds (35 females and 11 males) issued from two lines selected for a more (line R: 25 females and 10 males) or less (line A: 10 females and 1 male) clear circadian rhythm of feeding activity. The birds, were placed successively under three light schedules: constant darkness (DD), constant dim green light (LLdim) and constant bright light (LLbright). Schedules were changed every 2 weeks. Feeding activity was recorded continuously, analysed by autocorrelation and spectral analysis, and the ratio of the correlation coefficients and the area of the spectrum peak were used as indexes to quantify the clarity of the circadian rhythm. During the experiment, some birds showed gonadal development. Therefore, we analysed separately birds showing either a high or low degree of sexual development at the end of the experiment. In DD, 35 birds showed a circadian feeding rhythm with a mean period of 22.5 ± 0.1 h, whereas 11 birds showed an arrhythmic activity. In LLdim, 27 birds were rhythmic (22 birds R and 5 birds A), and in LLbright, only 3 birds showed a rhythmic circadian activity. For the R-line birds (for females and males), the rhythm clarity decreased in LLdim compared to DD, except for the not developed females. For the A-line birds (for females), the rhythm clarity of the immature birds increased in LLdim and that of the developed birds remained stable. In LLbright, circadian activity became arrhythmic. In LLdim, the active phases of 12 birds showed two main peaks, with mean periods of 22.7 h and 25.1 h, respectively. Therefore, constant light appeared to have an inhibitory effect on the expression of the circadian rhythm. We postulate that two hierarchically coupled oscillators could control circadian feeding activity, and arrhythmia in LLbright could be the results of internal desynchronization of the pacemakers.     

Effect of Light Intensity on Circadian Activity in Developing Japanese Quail

The effects of constant light on the expression of the circadian rhythm of feeding activity in Japanese quail, and in particular on the clarity of the rhythm were investigated. We used 46 4-week-old birds (35 females and 11 males) issued from two lines selected for a more (line R: 25 females and 10 males) or less (line A: 10 females and 1 male) clear circadian rhythm of feeding activity. The birds, were placed successively under three light schedules: constant darkness (DD), constant dim green light (LLdim) and constant bright light (LLbright). Schedules were changed every 2 weeks. Feeding activity was recorded continuously, analysed by autocorrelation and spectral analysis, and the ratio of the correlation coefficients and the area of the spectrum peak were used as indexes to quantify the clarity of the circadian rhythm. During the experiment, some birds showed gonadal development. Therefore, we analysed separately birds showing either a high or low degree of sexual development at the end of the experiment. In DD, 35 birds showed a circadian feeding rhythm with a mean period of 22.5 ± 0.1 h, whereas 11 birds showed an arrhythmic activity. In LLdim, 27 birds were rhythmic (22 birds R and 5 birds A), and in LLbright, only 3 birds showed a rhythmic circadian activity. For the R-line birds (for females and males), the rhythm clarity decreased in LLdim compared to DD, except for the not developed females. For the A-line birds (for females), the rhythm clarity of the immature birds increased in LLdim and that of the developed birds remained stable. In LLbright, circadian activity became arrhythmic. In LLdim, the active phases of 12 birds showed two main peaks, with mean periods of 22.7 h and 25.1 h, respectively. Therefore, constant light appeared to have an inhibitory effect on the expression of the circadian rhythm. We postulate that two hierarchically coupled oscillators could control circadian feeding activity, and arrhythmia in LLbright could be the results of internal desynchronization of the pacemakers.     

Regulation of PEP-Carboxylase by Biological Clock in a CAM Plant

The endogenous circadian rhythm in a crassulacean acid metabolism(CAM) plant Graptopetalum paraguayense was investigated. Phosphoenolpyruvatecarboxylase (PEP-C) takes two forms: the malate-sensitive dayform and the malate-insensitive night form. We monitored thestate of PEP-C by measuring the sensitivity to malate as a parameterof the circadian rhythm. We also measured vacuolar pH and malateconcentration, and contents of oxaloacetate, pyruvate and phosphoenolpyruvate(PEP). A free-running circadian oscillation was observed under continuousdim light (5 klux) after 12 h/12 h light/dark cycles at 20°C.The period of the rhythm was about 20 h. Under continuous light(18 klux), the rhythm was less clear but the length of the periodwas not affected. On the other hand, the rhythms of the vacuolarpH and the malate concentration were evident under the continuouslight, but were not clear under the continuous dim light. Therhythm disappeared in continuous darkness. The content of PEPchanged simultaneously with the transformation of PEP-C duringthe normal day-night cycles and under the continuous light,but stayed at a low level under the continuous dim light. Thisindicated that the transformation of PEP-C was not sufficientto maintain the rhythm in the carbon metabolism. Shift of the timing of the start or end of the dark period priorto the continuous illumination shifted the phase of the PEP-Crhythm without changing the period length significantly. At30°C, the rhythm of PEP-C was less clear, but the periodlength was not affected. These results suggest that the biological clock controls CO₂uptake and day-night CAM cycle through regulation of PEP-C transformation. (Received August 20, 1993; Accepted December 3, 1993)     

RHYTHMIC SETTLING BEHAVIOR IN PYRAMIMONAS PARKEAE (PRASINOPHYCEAE)1

Pyramimonas parkeae Norris et Pearson was examined for evidence of a settling rhythm while growing in laboratory culture. The alga settled rhythmically in a diet cycle when kept in a regularly alternating light-dark cycle. Cells moved out of suspension when settling and attached themselves to the sides and base of the culture vessel. Although rhythmic settling was inhibited in constant dim light (LL), it continued in constant darkness for 4 days with a period of approximately 24 h. The settling rhythm was temperature-compensated and could be reset by 6-h exposures to light. These observations demonstrate that the settling behavior of P. parkeae is controlled by an endogenous circadian oscillator. Regular low temperature pulses every 12 h removed the inhibition caused by LL and this points to the possible role of temperature changes as stimuli entraining the circadian settling rhythm of P. parkeae.     

Does the tsetse parturition rhythm have a circadian basis?

Abstract. Under an LD 12:12h photoregime at constant temperature, parturition in Glossina morsitans centralis is a gated event occurring late in the afternoon. When flies are switched to continuous light the rhythm quickly dampens, but its persistence for at least two 24-h cycles beyond the final scotophase suggests the rhythm has a circadian basis. A weak rhythm appears after 7 days of continuous light, perhaps in response to the daily disturbance caused by feeding. Return of the flies to LD 12:12h restores the rhythm after exposure to a single scotophase.     

Adult emergence rhythm of the egg-parasitoid Telenomus busseolae

The adult emergence rhythm of Telenomus busseolae, an egg parasitoid of Sesamia nonagrioides, was examined when parasitoids were exposed to different light-dark regimes. Most of the adult parasitoids emerged throughout the whole period of the photoperiodic cycle. Peak male emergence occurred 2–5 hours earlier than that of females. Adult emergence was asynchronous in continuous darkness or light. However, regimes of alternative light and dark phases such as L4:D20, L8:D16, L12:D12, L16:D8 and L20:D4 h generated a population rhythm with a period length of 24 hours. The peak of the emergence activity moves from the scotophase to the middle of the photophase with an increase of the photophase from 4 to 20 h. Rhythmical activity of adults was synchronised within 2 cycles when immature stages of parasitoid grow under continuous light conditions (LL) and then transferred to L12:D12. Moreover, emergence rhythm persisted and continued in a free-run with a period length of less than 24 hours by transferring a rhythmic culture from L12:D12 h to LL or RR (continuous red light) conditions, indicating the existence of a circadian rhythm. The ecological implications of the expression rhythm relate to better survival of the parasitoids.     

Circadian rhythms in the mating behavior of the cockroach, Leucophaea maderae

Mating behavior of small populations of virgin males and females of the cockroach Leucophaea maderae were continuously monitored via time-lapse video recording in controlled laboratory conditions. The time of onset of copulation was found to be rhythmic in a light cycle of 12 h light alternated with 12 h of darkness, with the peak of mating behavior occurring near the light to dark transition. This rhythm persisted in constant dim red illumination and constant temperature. In constant conditions, the period of the rhythm was slightly less than 24 h, with a peak of copulation during the late subjective day. These data demonstrated that mating behavior is gated by a circadian clock. When males and females were taken from light cycles that were 12 h out of phase, a bimodal rhythm was observed with one peak in the males' late subjective day and a second peak of equal amplitude in the late subjective day of females. The results indicated that circadian systems in both males and females contribute to the circadian rhythm in copulation. Bilateral section of the optic tracts (OTX) of both males and females abolished the rhythm, but the rhythm persisted when OTX females were paired with intact males or when OTX males were paired with intact females. Furthermore, when OTX males or OTX females were paired with intact animals that were 12 h out of phase, a bimodal rhythm was still observed. These results suggested that the circadian pacemaker in the optic lobes of both male and female cockroaches participates in the control of mating, but that a pacemaker outside the optic lobes is also likely involved. Finally, it was shown that the female's olfactory response (measured by electroantennogram) to components of the male sex pheromone exhibited a circadian rhythm, but the data suggested the peripheral olfactory rhythm is not likely to be involved in the rhythm of mating behavior.     

Reanalysis of Filter-Feeding Behavior of Caddis Fly (Brachycentrus) Larvae Reveals Masking and Circadian Rhythmicity

Data published on the subjectively determined presence or absence of circadian periodicity in feeding of Brachycentrus occidentalis Banks, an aquatic insect belonging to the order Trichoptera, were reanalyzed by statistical methods based on the cosine model. The 4h data for 5 days of synchronized environmental conditions followed by 5 days of constant conditions (using original values and purified values after removal of an 8h component attributed to the feeding schedule) were quantified for rhythm characteristics by the least-squares fit of cosines. A circadian rhythm was highly significant during synchronized conditions (light-dark and cycling water temperature) and in continuous light when water temperature continued to cycle. A circadian rhythm in the percentage of larvae feeding during continuous light and constant water temperature, which had not been observed by visual inspection in the original data, was detected in the unmasked data with a “free-running” period not overlapping 24.0h (95% confidence limits from 18.95h to 21.66h). Effects of masking brought about by manually feeding the larvae every 8h were statistically significant under both synchronized and constant environmental conditions. (Chronobiology International, 15(6), 595-606, 1998)     

Effects of light pulses on the locomotor activity rhythm of Orchestia montagui (Amphipoda,Talitridae)

Animals of the amphipod Orchestia montagui are kept in constant darkness with two short light pulses. One pulse is applied at the beginning of subjective night (around the dusk) and the other one at the end of subjective night (around the dawn). The pulse duration is estimated in the order of one or two hours around the dusk as well as the dawn. The locomotor activity rhythm was monitored in individual animals in summer under constant temperature. Results revealed that whatever the experimental conditions, under continuous or interrupted darkness by pulses, two endogenous components have been highlighted. In fact, Periodogram analysis showed the presence of ultradian and circadian periods around 12 and 24 h, respectively. The shortest circadian period and the most important inter-individual variability was observed under pulse of 2 h around the dusk with mean value equal to τ_DD+pulse = 24h38′ ± 4h34′. The activity profiles are in majority unimodal. Moreover, the most activity peak showed a slipping of its location from the middle of subjective night under constant darkness to the middle of subjective day under pulse. Globally, the locomotor activity rhythm of O. montagui was better defined under pulses and specimens were significantly more active under continuous darkness. Moreover, a great variability around the activity time was observed especially with pulse of 1 h.     

Characteristics of the circadian activity rhythm in common marmosets (Callithrix j. jacchus)

The circadian activity rhythm of the common marmoset, Callithrix j. jacchus was investigated by long-term recording of the locomotor activity of 15 individuals (5 males, 10 females) from 1.5 to 8 years old, both under constant illumination and under LD 12:12. The mean period of the spontaneous circadian rhythm was 23.2 ± 0.3 h. Neither sex-specific differences nor a systematic influence of light intensity on the spontaneous period were observed, but the period was dependent on the duration of the trial and on the age of the individual. Due to the short spontaneous period, in LD 12:12 there was a distinct advance of the activity phase with respect to the light time and a masking of the true onset of activity by the inhibitory direct effect of low light intensity during the dark time. After an 8 h delay shift of the LD 12:12, re-entrainment of the circadian activity rhythm required an average of 6.8 ± 0.7 days; the average re-entrainment time after an 8 h phase advance of the LD cycle was 8.6 ± 1.3 day. This directional effect is ascribed to characteristics of the phase-response curve. No ultradian components were observed, either in the LD-entrained or the free-running circadian activity rhythm.     

Light- and temperature-entrained circadian regulation of activity and mRNA accumulation of 1-aminocyclopropane-1-carboxylic acid oxidase in Stellaria longipes

Stem and leaf tissues of Stellaria longipes Goldie (prairie ecotype) exhibit circadian rhythmicity in the activity and mRNA abundance for 1-aminocyclopropane-1-carboxylic acid oxidase (EC 1.4.3). The steady-state mRNA levels and enzymatic activity levels fluctuated with a period of approximately 24 h and reached their maxima by the middle of the light phase and minima by the middle of the dark phase. The oscillations showed damping under constant light, constant dark and constant temperature conditions, indicating that the rhythm is entrained by an external signal. The results indicate that light/dark cycles have greater entraining effects than temperature cycles. A 15-min red light pulse, but not a blue light pulse, could reset rhythm in continuous darkness, suggesting the possible role of a red-light signal transduction pathway in the circadian regulation of 1-aminocyclopropane-1-carboxylic acid oxidase.Abbreviations ACC 1-aminocyclopropane-1-carboxylic acid - DD continuous dark - LD light-dark - LL continuous light - ZT Zeitgeber time (start of light period for circadian entrainment) This study was supported by operating grants to C.C.C., and D.M.R. from the Natural Sciences and Engineering Research Council of Canada.The authors gratefully acknowledge the award of a Bettina Bahlsen memorial Graduate Scholarship by University of Calgary to A.K. We are grateful to Dr. M.M. Moloney for allowing the use of his laboratory facilities.