Effects of fasting on thermoregulatory processes and the daily oscillations in rats

To investigate the mechanism involved in the reduction of body core temperature (T(core)) during fasting in rats, which is selective in the light phase, we measured T(core), surface temperature, and oxygen consumption rate in fed control animals and in fasted animals on day 3 of fasting and day 4 of recovery at an ambient temperature (T(a)) of 23 degrees C by biotelemetry, infrared thermography, and indirect calorimetry, respectively. On the fasting day, 1) T(core) in the light phase decreased (P < 0.05) from the control; however, T(core) in the dark phase was unchanged, 2) tail temperature fell from the control (P < 0.05, from 30.7 +/- 0.1 to 23.9 +/- 0.1 degrees C in the dark phase and from 29.4 +/- 0.1 to 25.2 +/- 0.2 degrees C in the light phase), 3) oxygen consumption rate decreased from the control (P < 0.05, from 24.37 +/- 1.06 to 16.24 +/- 0.69 ml. min(-1). kg body wt(-0.75) in the dark phase and from 18.91 +/- 0.64 to 14.00 +/- 0.41 ml. min(-1). kg body wt(-0.75) in the light phase). All these values returned to the control levels on the recovery day. The results suggest that, in the fasting condition, T(core) in the dark phase was maintained by suppression of the heat loss mechanism, despite the reduction of metabolic heat production. In contrast, the response was weakened in the light phase, decreasing T(core) greatly. Moreover, the change in the regulation of tail blood flow was a likely mechanism to suppress heat loss.     

Combinations of bright light, scheduled dark, sunglasses, and melatonin to facilitate circadian entrainment to night shift work

Various combinations of interventions were used to phase-delay circadian rhythms to correct their misalignment with night work and day sleep. Young participants (median age = 22, n = 67) participated in 5 consecutive simulated night shifts (2300 to 0700) and then slept at home (0830 to 1530) in darkened bedrooms. Participants wore sunglasses with normal or dark lenses (transmission 15% or 2%) when outside during the day. Participants took placebo or melatonin (1.8 mg sustained release) before daytime sleep. During the night shifts, participants were exposed to a moving (delaying) pattern of intermittent bright light (approximately 5000 lux, 20 min on, 40 min off, 4-5 light pulses/night) or remained in dim light (approximately 150 lux). There were 6 intervention groups ranging from the least complex (normal sunglasses) to the most complex (dark sunglasses + bright light + melatonin). The dim light melatonin onset (DLMO) was assessed before and after the night shifts (baseline and final), and 7 h was added to estimate the temperature minimum (Tmin). Participants were categorized by their amount of reentrainment based on their final Tmin: not re-entrained (Tmin before the daytime dark/sleep period), partially re-entrained (Tmin during the first half of dark/sleep), or completely re-entrained (Tmin during the second half of dark/ sleep). The sample was split into earlier participants (baseline Tmin < or = 0700, sunlight during the commute home fell after the Tmin) and later participants (baseline Tmin > 0700). The later participants were completely re-entrained regardless of intervention group, whereas the degree of re-entrainment for the earlier participants depended on the interventions. With bright light during the night shift, almost all of the earlier participants achieved complete re-entrainment, and the phase delay shift was so large that darker sunglasses and melatonin could not increase its magnitude. With only room light during the night shift, darker sunglasses helped earlier participants phase-delay more than normal sunglasses, but melatonin did not increase the phase delay. The authors recommend the combination of intermittent bright light during the night shift, sunglasses (as dark as possible) during the commute home, and a regular, early daytime dark/sleep period if the goal is complete circadian adaptation to night-shift work.     

Light/dark patterns of interleukin-6 in relation to the pineal hormone melatonin in patients with acute myocardial infarction

AIMS: Atherosclerosis is a chronic disease that, from its origin to its ultimate complications, involves inflammatory cells, inflammatory proteins, and inflammatory responses from vascular cells. It has been demonstrated that cytokine activities are under neuroendocrine control, in part exerted by the pineal gland through the circadian secretion of its main product melatonin. Melatonin is mainly released during the night, but the precise relationship between melatonin and the light/dark rhythm of interleukin-6 in patients with acute myocardial infarction is still unclear. METHODS AND RESULTS: The study included 60 patients diagnosed with acute myocardial infarction and 60 healthy volunteers whose venous blood samples were collected at 09:00 h (light period) and 02:00 h (dark period). Our results demonstrate that interleukin-6 concentrations presented a light/dark pattern with mean serum concentrations being higher in the acute myocardial infarction group than in the control group (101.26 +/- 13.43 and 52.67 +/- 7.73 pg/ml at 02:00 h, 41.93 +/- 5.90 and 22.98 +/- 4.49 pg/ml at 09:00 h, respectively, p < 0.05). Differences in the day/night changes in melatonin levels in control subjects (48.19 +/- 7.82 at 02:00 h, 14.51+/- 2.36 at 09:00 h, pg/ml) and acute myocardial infarction patients (25.97 +/- 3.90 at 02:00 h, 12.29 +/- 4.01 at 09:00 h, pg/ml) (p < 0.05) were a result of a reduced nocturnal elevation of melatonin in the acute myocardial infarction group. CONCLUSIONS: The current findings suggest that the circadian secretion of melatonin may be responsible at least in part for light/dark variations of endogenous interleukin-6 production in patients with acute myocardial infarction. In this study, the melatonin seems to have an anti-inflammatory effect.     

Correlation between the Circadian Rhythm of Resistance to Extreme Temperatures and Changes in Fatty Acid Composition in Cotton Seedlings

Fluctuations in fatty acid composition were examined in cotton (Gossypium hirsutum L. cv Deltapine 50) leaves during light-dark cycles of 12:12 h and under continuous light and were correlated to the rhythmic changes in chilling (5[deg]C) resistance (CR) and heat (53[deg]C) resistance (HR). The chilling-resistant and chilling-sensitive phases developed in the dark or the light period, respectively, and this rhythm persisted under continuous light for three cycles. The heat-resistant phase developed in the light period and an additional peak of HR occurred in the middle of the dark period. Under continuous light, only one peak of HR developed, lasting from the middle of the subjective night to the middle of the subjective day. The amounts of palmitic and oleic acids were constant during the light-dark cycle and under continuous light, but those of linoleic and linolenic acids fluctuated, attaining a high level in the middle of the dark period or the subjective night, and a low level in the middle of the light period or the subjective day. A low temperature of 20[deg]C induced CR and affected changes in fatty acid composition similar to those that occurred during the daily CR phase. A high temperature of 40[deg]C induced HR but did not affect changes in fatty acid composition. The results in their entirety show that the CR that develops rhythmically as well as the low-temperature-induced CR coincide with increased levels of polyunsaturated fatty acids. No correlation is found between changes in fatty acid composition and the HR that develops rhythmically or the high-temperature-induced HR.     

Effects of light intensity and restraint on dark-pulse-induced circadian phase shifting during subjective night in Syrian hamsters

Dark pulses presented on a background of constant light (LL) result in phase advances during midsubjective day and early subjective night, and phase delays during late subjective night, as shown in the dark-pulse phase response curve. In hamsters, the phase-shifting effects of dark pulses are thought to be mediated by increased activity, as previous studies have shown that restraining animals during dark pulses blocks the phase shifts observed in midsubjective day and late subjective night. This study focuses on dark-pulse-induced phase shifting during early subjective night, examining the influence of both LL intensity and restraint on the magnitude of these phase shifts. Syrian hamsters were maintained in LL of four different illumination levels (1, 10, 100, or 600 lux) and periodically presented with 6-h pulses (dark pulse alone, restraint alone, or dark pulse plus restraint) beginning at circadian time 11. Phase advances were observed in response to dark pulses alone, and the magnitude of these shifts was dependent on background illumination, with significantly larger advances seen under higher intensities. No relationship was found between the amount of activity displayed during dark pulses and phase shift magnitude. Six-hour periods of restraint resulted in phase delays, the magnitude of which was also dependent on background illumination. Restraining hamsters during dark pulses reduced the magnitude of phase advances, but the extent of this reduction could be predicted from the additive effects of the dark-pulse-alone and restraint-alone conditions. These results indicate that the phase-shifting effects of dark pulses during early subjective night are not mediated by behavioral activation and may instead reflect a mirror image of the phase-delaying effects of light pulses at this phase.     

Diurnal behavioral and endocrine effects of chronic shaker stress in mice

Experiments were performed in C57BL/6J male mice to determine 1) light/dark effects of acute and chronic shaker stress on open field behavioral patterns and 2) light/dark effects of chronic stress on plasma corticosterone and oxytocin. Shaker stress was applied acutely (15 min) or chronically (3 or 7 days). Mice were tested in the open field in the light or dark phase of the circadian cycle. For the endocrine study, mice were exposed to 3 days of intermittent shaker stress and sacrificed after the last stress event (09:00 or 19:00 h). Acute or chronic shaker stress had no significant effects on intensity of motor activity and rearing of mice tested under either light condition. Mice tested in the dark phase had higher motor activity and exhibited lower anxiety-like behavior as expressed by central zone activities and had higher emotionality as expressed by increased defecation. Chronic stress increased corticosterone with a greater absolute increase in the dark period. However, the percentage stress-induced increase was not different between the day and night periods. The oxytocin response to stress was observed only during the light phase with no change seen at dark phase. These results show that there is a marked difference in the light/dark pituitary stress response with no alteration in stress induced behavioral changes. They also suggest that there are circadian interactions in the endocrine stress axis that are without consequences for open field behavior.     

Entrainment of daily serum gonadotropin cycles in the goldfish to photoperiod, feeding, and daily thermocycles

Female fish were kept under 16L:8D/20 degrees C in November and April and the onset of light and/or feeding times were shifted by several hours in the experimental groups. Photoperiod and feeding entrained significant fluctuations in serum gonadotropin hormone (GTH) levels when the onset of light and the first daily feeding were 4 hr apart, but not when they were 10 hr apart. Fish were subjected to 16L:8D for 14-16 days in February, and to either a constant warm (20 degrees C) or a diurnal sinusoidal (12-20 degrees C) temperature regime, the warmth being imposed during photophase or scotophase. While relatively high, uniform serum GTH levels were found throughout the 24-hr period in fish subjected to constant warmth, warm temperature during the day promoted fluctuations in serum GTH levels, and warmth during night resulted in relatively low, uniform serum GTH levels.     

Circadian rhythm of heat resistance in cotton seedlings: synthesis of heat-shock proteins.

Cotton (Gossypium hirsutum L. cv. Deltapine 50) seedlings grown under light-dark cycles of 12:12 h at 33 degrees C showed rhythmic changes in their resistance to heat shock of 53 degrees C for 40 min. The resistance was maximal at the middle of the light period and declined toward the end of the light period. One more peak of resistance developed in the middle of the dark period and declined toward the end of the dark period. Rhythmic changes in heat resistance persisted under continuous light for 3 cycles, indicating a circadian control. Under continuous light only one phase of resistance developed, lasting from the middle of the subjective night to the middle of the subjective day. The major heat shock proteins (HSPs) synthesized upon 30-min exposure to 40 degrees C, 49 degrees C or 53 degrees C were of 115, 89, 73, and 19 kDa. Their rate of synthesis depended on the inducing temperature, on previous exposure to high temperature and on the time in the light-dark cycle. The time dependency of the induction of certain HSPs persisted under continuous light, indicating a circadian control. No positive correlations was found between the rhythmic changes in heat resistance and the rhythmic changes in the synthesis of any HSP.     

Diel variation of the cellular carbon to nitrogen ratio of Chlorella autotrophica (Chlorophyta) growing in phosphorus‐ and nitrogen‐limited continuous cultures

We investigated the relationship between daily growth rates and diel variation of carbon (C) metabolism and C to nitrogen (N) ratio under P‐ and N‐limitation in the green algae Chlorella autotrophica. To do this, continuous cultures of C. autotrophica were maintained in a cyclostat culture system under 14:10 light:dark cycle over a series of P‐ and N‐limited growth rates. Cell abundance, together with cell size, as reflected by side scatter signal from flow cytometric analysis demonstrated a synchronized diel pattern with cell division occurring at night. Under either type of nutrient limitation, the cellular C:N ratio increased through the light period and decreased through the dark period over all growth rates, indicating a higher diel variation of C metabolism than that of N. Daily average cellular C:N ratios were higher at lower dilution rates under both types of nutrient limitation but cell enlargement was only observed at lower dilution rates under P‐limitation. Carbon specific growth rates during the dark period positively correlated with cellular daily growth rates (dilution rates), with net loss of C during night at the lowest growth rates under N‐limitation. Under P‐limitation, dark C specific growth rates were close to zero at low dilution rates but also exhibited an increasing trend at high dilution rates. In general, diel variations of cellular C:N were low when dark C specific growth rates were high. This result indicated that the fast growing cells performed dark C assimilation at high rates, hence diminished the uncoupling of C and N metabolism at night.     

Discrimination Processes and Shifts in Carboxylation during the Phases of Crassulacean Acid Metabolism

The magnitude and extent of Crassulacean acid metabolism (CAM) activity in two Clusia species was manipulated to investigate the regulation of the distinct CAM phases. First, in response to leaf-air vapor pressure deficit at night, changes in leaf conductance altered on-line carbon-isotope discrimination throughout the theoretical range for dark CO2 uptake during CAM. These ranged from the limit set by phosphoenolpyruvate carboxylase (PEPc) (-6[per mille (thousand) sign], [delta]13C equivalent of -2[per mille (thousand) sign]) to that imposed by diffusion limitation (+4[per mille (thousand) sign], [delta]13C equivalent of -12[per mille (thousand) sign]), but the lowest carbon-isotope discrimination occurred when P[square root]pa was only 0.7. Second, when the availability of external or internal sources of CO2 was reduced for both field- and greenhouse-grown plants, CO2 uptake by day via PEPc during phase II largely compensated. Third, by reducing the dark period, plants accumulated low levels of acidity, and CO2 uptake occurred throughout the subsequent light period. Discrimination switched from being dominated by PEPc (phase II) to ribulose 1,5-bisphosphate carboxylase/oxygenase (phase III), with both enzymes active during phase IV. Under natural conditions, photochemical stability is maintained by extended PEPc activity in phase II, which enhances acid accumulation and delays decarboxylation until temperature and light stress are maximal at midday.     

Environmental and genetic effects on circadian clock-regulated gene expression in Arabidopsis.

Expression patterns of the cold-circadian rhythm-RNA binding (CCR) and chlorophyll a/b binding (CAB) protein genes have circadian rhythms with phases that are different from each other and are affected differently by cold (4 degrees C) treatment. Cycling of CCR and CAB RNA levels was observed in Arabidopsis seedlings grown for 5 days at 4 degrees C under a light/ dark photoperiod, although the cycling had reduced amplitude compared with normal growth conditions (20 degrees C). CCR RNA levels were elevated in the cold, whereas CAB RNA levels were reduced in the cold relative to levels in control seedlings. Cold pulses (4 degrees C for 12 or 20 hr) under continuous light affected the rhythms of CCR and CAB RNA levels in similar ways. The 12-hr cold pulse caused a 4-hr phase delay in both rhythms, whereas the 20-hr cold pulse resulted in a 12-hr phase delay in both rhythms. The timing of CAB expression 1 (toc1) mutation shortened the period of the CCR rhythm, matching previous results for the regulation of the CAB-luciferase (CAB-luc) transgene in this mutant. The results suggest that CCR and CAB share clock machinery but are regulated by downstream components that are affected differently by the cold. Also, the circadian clock regulating these genes in Arabidopsis has a cold-sensitive phase under continuous light conditions.     

Dark-phase light contamination disrupts circadian rhythms in plasma measures of endocrine physiology and metabolism in rats

Dark-phase light contamination can significantly disrupt chronobiologic rhythms, thereby potentially altering the endocrine physiology and metabolism of experimental animals and influencing the outcome of scientific investigations. We sought to determine whether exposure to low-level light contamination during the dark phase influenced the normally entrained circadian rhythms of various substances in plasma. Male Sprague-Dawley rats (n = 6 per group) were housed in photobiologic light-exposure chambers configured to create 1) a 12:12-h light:dark cycle without dark-phase light contamination (control condition; 123 μW/cm(2), lights on at 0600), 2) experimental exposure to a low level of light during the 12-h dark phase (with 0.02, 0.05, 0.06, or 0.08 μW/cm(2) light at night), or 3) constant bright light (123 μW/cm(2)). Dietary and water intakes were recorded daily. After 2 wk, rats underwent 6 low-volume blood draws at 4-h intervals (beginning at 0400) during both the light and dark phases. Circadian rhythms in dietary and water intake and levels of plasma total fatty acids and lipid fractions remained entrained during exposure to either control conditions or low-intensity light during the dark phase. However, these patterns were disrupted in rats exposed to constant bright light. Circadian patterns of plasma melatonin, glucose, lactic acid, and corticosterone were maintained in all rats except those exposed to constant bright light or the highest level of light during the dark phase. Therefore even minimal light contamination during the dark phase can disrupt normal circadian rhythms of endocrine metabolism and physiology and may alter the outcome of scientific investigations.     

Effect of systemic hypoxia and reoxygenation on electrical stability of the rat myocardium: chronophysiological study

The aim of the study was to determine the dependence of changes in the electrical stability of the heart on the light-dark cycle (LD cycle) in disorders of pulmonary ventilation. The ventricular arrhythmia threshold (VAT) was measured in female Wistar rats (adaptation to the light regime 12:12 h, ketamine/xylazine anesthesia 100 mg/15 mg/kg, i.m., open chest experiments). The conditions of the normal artificial ventilation and reoxygenation were V(T) = 1 ml/100 g, respiratory rate 40 breaths/min, hypoventilation V(T) = 0.5 ml/100 g, respiratory rate 20 breaths/min. The animals (n=11 light group; n=19 dark group) were subjected to 20 min hypoventilation followed by 20 min reoxygenation. The control prehypoventilatory VAT differences were not found between the light (1.90+/-0.84 mA) and dark (1.88+/-0.87 mA) part of the day. Artificial hypoventilation changed the VAT values in light and dark part of the day differently. While during the light period, the average VAT values in most animals (90.9 %) were significantly decreased (1.29+/-0.59 vs. 1.90+/-0.84 mA control, p<0.05), during the dark part these values showed either significant increase (63.2 %) (2.23+/-0.77 vs. 1.48+/-0.39 mA, p<0.005) or a slight non-significant decrease (36.8 %) (2.18+/-0.89 vs. 2.54+/-0.99 mA). Reoxygenation returned the VAT values to the level before hypoventilation by an increase of the VAT (81.8 %) in the light part of day and by decrease of the VAT (68.4 %) in the dark part of the day. It is concluded that 1) in hypoventilation/reoxygenation model, the significant higher average VAT values are in the dark part of the day vs. the light one, 2) rat hearts are more resistant to systemic hypoxia and reoxygenation in the dark part of day, and 3) proarrhythmogenic effect of the systemic hypoxia is only seen in the light part of the day.     

Diurnal variations in tonic pain reactions in mice

Diurnal changes in the behavioural reactions to subcutaneous formalin injection (20 microl, 1%) into the dorsum of an hindpaw were examined in female CBA/J mice aged 70-75 days, maintained in a 12/12 dark/ light cycle (light on at 07.00 h; light off at 19.00 h). Mice showed higher pain scores, as expressed by the amount of time spent licking the injected paw and by the number of flinching episodes, when tested under red light at the beginning of the dark phase (19.00-22.00: Dark group) than when tested either under white or red light at the beginning of the light phase of the diurnal cycle (7.00-10.00). The increases in pain reactions at night were found both during the first (0-10 min) and the second (11-55 min) phase of the behavioural response to formalin injection. They were not due to aspecific increases in motor behaviour, since self-grooming actually decreased in the Dark group during the second phase of the response, and the amount of locomotor activity after the injection was similar to, or lower than, that found in mice tested in the morning under white or red light, respectively. In another group of female CBA/J mice tested in the hotplate apparatus (at a temperature of 52 degrees), paw-lick latencies were significantly higher in mice tested at dark during the night, whereas jump (escape) latencies were higher in the morning. These results demonstrate different diurnal variations in the reactions to brief or prolonged noxious stimulation in mice, with greater responses to tonic pain at the onset of the dark phase.     

Variations in plasma melatonin levels of the rainbow trout (Oncorhynchus mykiss) under various light and temperature conditions

Daily variations in plasma melatonin levels in the rainbow trout Oncorhynchus mykiss were studied under various light and temperature conditions. Plasma melatonin levels were higher at mid-dark than those at mid-light under light-dark (LD) cycles. An acute exposure to darkness (2 hr) during the light phase significantly elevated the plasma melatonin to the level that is comparable with those at mid-dark, while an acute exposure to a light pulse (2 hr) during the dark phase significantly suppressed melatonin to the level that is comparable with those at mid-light. Plasma melatonin kept constantly high and low levels under constant darkness and constant light, respectively. No circadian rhythm was seen under both conditions. When the fish were subjected to simulative seasonal conditions (simulative (S)-spring: under LD 13.1:10.9 at 13 degrees C; S-summer: under LD 14.3:9.7 at 16.5 degrees C; S-autumn: under LD 11.3:12.7 at 13 degrees C; S-winter: under LD 10.1:13.9 at 9 degrees C), melatonin levels during the dark phase were significantly higher than those during the light phase irrespective of simulative seasons. The peak melatonin level in each simulative season significantly correlated with temperature but not with the length of the dark phase employed. In addition, the peak melatonin level in S-autumn was significantly higher than those in S-spring although water temperature was the same under these conditions. These results indicate that the melatonin rhythm in the trout plasma is not regulated by an endogenous circadian clock but by combination of photoperiod and water temperature.     

Day-night variations in plasma melatonin and arginine vasotocin concentrations in chronically cannulated flounder (Platichthys flesus).

Chronically catheterised, free swimming flounder (Platichthys flesus) have been used in experiments examining the day-night variations in circulating levels of melatonin (Mel) and arginine vasotocin (AVT). Under normal photoperiod (16 h light/8 h dark) serial blood samples taken from individual fish demonstrated a Mel rhythm with daytime levels at 09.00 and 15.00 h (238+/-14 and 179+/-12 fmol x ml(-1), respectively) lower than those at 23.00 h (1920+/-128 fmol x ml(-1)). Maintenance of fish in 24-h light abolished the light/dark Mel rhythm and circulating levels were comparable to those measured during the day in fish under normal photoperiod illumination. In fish maintained under 24 h dark, although a daily rhythm was still apparent, at the time when it would be normally dark, plasma Mel concentration was reduced and at times when it would be normally light, levels were higher than in fish maintained under normal light/dark illumination. Plasma AVT concentrations were higher in fish during the day (4.4+/-0.8 fmol x ml(-1)) than those at night (1.5+/-0.4 fmol x ml(-1)), the opposite to that seen with Mel. During acute study infusion of AVT resulted in reduced levels of plasma Mel, although this did not achieve statistical significance. Infusion of Mel did not alter circulating AVT concentration.     

Repercussions of anterior hypothalamic lesions on nycthemeral variations of plasma thyroxine and corticosterone in the pigeon

Plasma thyroxine and corticosterone levels were determined by competitive protein binding assay, at 3 hr intervals, throughout the photoperiod. Pigeons were kept in controlled environment (21 +/- 1 degree C; 14L6-20: 10D). Intact controls exhibited low thyroxine (T4) and corticosterone (B) levels for the light phase of the photoperiod. Values were rising during the night, up to a peak at 03 hr. Electrolytic lesions were placed bilaterally in either the nucleus anterior medialis hypothalami or the n. preopticus, or the n. supraopticus. Circadian rhythms of both T4 and B were markedly altered in all lesioned pigeons, with a shift of very high T4 values to the morning times and a complete disorganization of B patterns, with very heterogeneous values. The possibility is raised that anterior hypothalamic formations participate in the endogenous oscillator circuitry in birds.     

Functional similarity in relation to the external environment between circadian behavioral and melatonin rhythms in the subtropical Indian weaver bird

The present study investigated whether the circadian oscillators controlling rhythms in activity behavior and melatonin secretion shared similar functional relationship with the external environment. We simultaneously measured the effects of varying illuminations on rhythms of movement and melatonin levels in Indian weaver birds under synchronized (experiment 1) and freerunning (experiment 2) light conditions. In experiment 1, weaverbirds were exposed to 12h light: 12h darkness (12L:12D; L = 20 lx, D = 0.1 lx) for 2.5 weeks. Then, the illumination of the dark period was sequentially enhanced to 1-, 5-, 10-, 20- and 100 lx at the intervals of about 2 to 4 weeks. In experiment 2, weaver birds similarly exposed for 2.5 weeks to 12L:12D (L = 100 lx; D = 0.1 lx) were released in constant dim light (LL(dim), 0.1 lx) for 6 weeks. Thereafter, LL(dim) illumination was sequentially enhanced to 1-, 3- and 5 lx at the intervals of about 2 weeks. Whereas the activity of singly housed individuals was continuously recorded, the plasma melatonin levels were measured at two time of the day, once in each light condition. The circadian outputs in activity and melatonin were phase coupled with an inverse phase relationship: melatonin levels were low during the active phase (light period) and high during the inactive phase (dark period). This phase relationship continued in both the synchronized and freerunning states as long as circadian activity and melatonin oscillators subjectively interpreted synchronously the daily light environment, based on illumination intensity and/or photophase contrast, as the times of day and night. There were dissociations between the response of the activity rhythms and melatonin rhythms in light conditions when the contrast between day and night was much reduced (20:10 lx) or became equal. We suggest that circadian oscillators governing activity behavior and melatonin secretion in weaverbirds are phase coupled, but they seem to independently respond to environmental cues. This would probably explain the varying degree to which the involvement of pineal/melatonin in regulation of circadian behaviors has been found among different birds.