The internal synchronization of five circadian functions of the rabbit

Five different physiological functions of the rabbit (hard faeces and urine excretion, food and water intake and locomotor activity) were registered during LD 12:12 and during continuous light conditions (LL).

(1) In LD 12:12 a strong synchronization of the five parameters existed. The minima of all functions consistently occurred during the hours of light. The nocturnal percentage of overall 24-hr events was increased significantly in 'hard faeces excretion' (66±8 (S.D.) %), 'water intake' (64±15 (S.D.) %) and 'urine excretion' (58±10 (S.D.) %). The nocturnal percentage of locomotor activity was significantly increased during the dark-hours in 9 out of 14 animals. In the other five individuals prominent peaks were present even during the photoperiod. On the average of all 14 animals 5S±13 (S.D.) % of the 24 hr events of locomotor activity occurred during the night. Despite a trough during the cessation of hard faeces excretion the events of food intake were not elevated significantly during the dark hours.

(2) During LL the synchronization of the five functions within each animal persisted during the complete 90-day LL period. A free-running circadian rhythm with-: = 24.8±0.5 (S.D.) hr was present in the four rabbits kept in LL conditions within 5-16 days after the withdrawal of the zeitgeber.

(3) In addition to the circadian period the power spectrum analysis of data obtained during LD 12:12 revealed significant ultradian periods of an average period length of 11,6 hr (hard faeces and urine excretion), 8 hr (food and water intake, locomotor activity) and 4 hr (food intake, locomotor activity). During the free-run ultradian periods of 8 and 3.2-4.2 hr were significant in almost all parameters.

(4) During LL the level of locomotor activity was reduced for 13±16 (S.D.) %, the events of food intake were increased for 17±12 (S.D.) %.

(5) The reinserted LD 12:12 zeitgeber re-entrained the circadian rhythms within 25-45 days.

(6) These results provided evidence of a predominant nocturnality of the rabbits under investigation.     

Plasma Corticosterone, Motor Activity and Metabolic Circadian Patterns in Streptozotocin-Induced Diabetic Rats

Experiments were conducted in male rats to study the effects of streptozotocin-induced diabetes on circadian rhythms of (a) plasma corticosterone concentrations; (b) motor activity; and (c) metabolic patterns. Animals were entrained to LD cycles of 12: 12 hr and fed ad libitum.

A daily rhythm of plasma corticosterone concentrations was found in controls animals with peak levels at 2400 hr and low values during the remaining hours. This rhythm was statistically confirmed by the cosinor method and had an amplitude of 3.37μg/100 ml and the acrophase at 100 hr. A loss of the normal circadian variation was observed in diabetic animals, with a nadir at the onset of light period and high values throughout the remaining hours; cosinor analysis of these data showed no circadian rhythm, delete and a higher mean level than controls.

As expected, normal rats presented most of their motor activity during the dark period with 80+ of total daily activity; the cosinor method demonstrated a circadian rhythm with an amplitude of 60+ of the mean level and the acrophase at 0852 hr. Both diabetic and control rats showed a similar activity during the light phase, but diabetic animals had less activity than controls during the night and their percentage of total daily activity was similar in both phases of the LD cycle (50+ for each one). With the cosinor method we were able to show the persistence of a circadian rhythm in the motor activity of diabetic rats, but with a mesor and amplitude lower than in controls (amplitude rested at 60+ of the mean level) and its acrophase advanced to 0148 hr.

The metabolic activity pattern of diabetic rats also changed: whereas controls showed a greater metabolic activity during the night (70+ food; 82+ water; 54+ urine; 67+ faeces), diabetics did not show differences between both phases of the LD cycle. Water ingested and urine excreted by the diabetic group were higher than normal during light and dark periods; food consumed and faeces excreted were higher than controls only in the light phase.

These data suggest that alterations in circadian rhythms of plasma corticosterone and motor activity are consecutive to the loss of the feeding circadian pattern, due to polyphagia and polydipsia showed by these animals, which need to extend intakes during the light and dark phases.     

Food-deprivation-induced phase shifts in Sminthopsis macroura froggatti

Past research has shown that there is a circadian oscillator in laboratory rats that is entrained by restricted feeding schedules. However, in laboratory rats at least, the light-dark (LD) cycle is the dominant zeitgeber in the entrainment of wheel-running activity rhythms. Given that dasyurid marsupials are predominantly carnivorous, the episodic intake of food in the wild and the high nutritive content of that food suggest that food may be an important zeitgeber in these species. Twelve Sminthopsis macroura froggatti were presented with a daily meal at 0900 hr under an LD 12:12 cycle with lights-on at 0600 hr for 37 days. Activity in anticipation of the meal was observed in most animals. Following this, all animals were exposed to periods of 12-18 days ad lib. food interspersed with 3-day periods of deprivation--a technique used previously to demonstrate persistent meal-associated rhythms. The meal-associated activity rhythms previously observed in rats during the 3-day deprivation period were not seen, but the 3-day deprivation period produced large phase-shifts in the activity rhythms of several S.m. froggatti. It is concluded that meal feeding does not dominate the LD cycle in entraining dasyurid marsupials, but that the frequent observation of phase shifts suggests a different and, perhaps, stronger role for food intake in biological rhythmicity than has been observed previously in laboratory rats.     

Circadian organization of running-wheel activity, food intake and drinking of old male rats

Circadian rhythms of running-wheel activity, food intake and drinking were monitored in old male rats of Long-Evans strain over 22 months of age in both entrained (light:dark 12:12, LD) and free running condition (continuous illumination, LL) and were compared with those of young adult male rats of 3.5 to 6.5 months of age. Twenty-four hour distribution of running activity, feeding events and licking events of young rats as well as old rats showed bi- or tri-modal patterns during the 12 hr dark period of the LD schedule. In the light period, 2 out of 8 old rats, 6 out of 10 old rats and 1 out of 6 old rats had 1 or 2 medium or high peaks in running activity, feeding events and licking events, respectively, leading to equal distribution between the dark and light period. In the LD schedule, old rats showed a decrease in running-wheel activity, its patterns and power spectra, a decrease in feeding events and its power spectra in 6 rats which lost circadian rhythms and increase in feeding events and its power spectra in 4 rats which still showed circadian rhythms and increase in licking events. LL suppressed running-wheel activity, its patterns and power spectra, licking events and its power spectra and feeding events in young rats. However, LL could suppress only feeding events of 4 rats which still showed circadian rhythms and licking events and its spectral level in old rats. The possible causes of decreased response to LL in old rats and its implication are discussed.     

The effect of fasting and refeeding on temperature preference,activity and growth of roach, <Emphasis Type="Italic">Rutilus rutilus</Emphasis>

Most fish species are regularly subjected to periods of starvation during which a reduction of energy turnover might be favourable for the animal. This reduction of energy flux may be achieved by changes in thermal behaviour and/or swimming activity. We investigated such behavioural changes during starvation and subsequent refeeding in roach, Rutilus rutilus, with respect to energetic benefits and growth maximisation. Roach, acclimated to a wide range of temperatures (4, 12, 20, 24, 27 and 30 °C), were fed to excess, subjected to 3 weeks of starvation and subsequently refed in order to determine the temperature dependence of feeding rates, growth rates and conversion efficiency (K₁) under control conditions and during compensatory growth. When exposed to a thermal gradient, control animals preferentially selected a temperature of 26.8ǂ.9 °C, which is in the range of the optimal temperatures for feeding, growth and conversion efficiency. Starving fish showed a distinct circadian pattern of the mean selected temperature (MST). They migrated to cooler water in the dark (MST_dark=22.8ǃ.1 °C) but returned to warmer water during daytime. This behaviour may be regarded as a trade-off between the potentially higher food density in warmer water areas and the energetic benefit of selecting cooler water patches. The circadian pattern of MST was gradually abandoned upon refeeding and control values were reached again after 3 weeks. Energetically more effective than behavioural hypothermia was the reduction of swimming activity. During starvation, activity peaks were slightly lower than under control conditions and mean daily activity decreased by about 50%. Swimming velocity, however, was not affected by feeding regime. After a period of starvation fish showed compensatory growth at all temperatures, even below 12 °C, where these animals normally do not grow. This suggests that after a period of starvation the critical temperature for growth shifts to lower values.     

Digestive tract cell proliferation and food consumption patterns of Ha/ICR mice

The relationship between the daily pattern of food consumption and the proliferation rate of the oesophagus, stomach, forestomach, small intestine and colon of Ha/ICR mice was examined. Proliferative activity was determined by [3H]TdR incorporation on a wet weight tissue basis, along with selective counting of labelled nuclei. Under conditions of ad libitum feeding with a 12 hr light cycle (lights on at 0600) mice eat most of their food during the dark period. A distinct circadian rhythm was observed in the oesophagus, stomach, forestomach and colon with the peak of [3H]TdR incorporation between 0400 and 0600 and the nadir between 1600 and 1800. Although a circadian fluctuation was observed in the small intestine, its amplitude was much less than in other areas. This rhythmic change in proliferation rate could be phase shifted by allowing the mice to feed only between 0800 and 1600 for 14 days. Under these conditions the peak in proliferative activity occurred between 1800 and 2000. Fasting reduced the daily level of proliferative activity in all of the digestive tract sites studied, and for all areas except the oesophagus greatly reduced or eliminated the circadian fluctuation. The forestomach and colon were the most influenced by fasting with 24 hr [3H]TdR incorporation reduced to 30-40% of the control value. Refeeding following a 48 hr fast produced a rapid increase in proliferative activity peaking at levels well above the control value at 16 hr after the onset of refeeding. The major exception to this was the small intestine which slowly returned to the control value during the first 24 hr. Partial refeeding produced a diminished refeeding response. Once the normal pattern of food consumption was re-established following refeeding the normal proliferative fluctuations were again observed.     

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

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

Ultradian and circadian compartmentalization of respiratory and metabolic exchanges in small laboratory vertebrates

Carbon dioxide emission (VCO2) taken as an index of respiratory and metabolic exchanges, was continuously recorded during 4-30 consecutive days in 100 quail, 87 chicks, 347 rats, 665 mice and 70 guinea-pigs which were under controlled environmental parameters. Harmonic analysis, fast Fourier transform, chi-square periodograms, peak and trough intervals were computed with VCO2 values obtained with CO2 concentrations sampled every 20 min on the CO2 recordings. In LD 12:12 alternation, circadian rhythms were observed in all quail, chicks, rats and mice, but only in 80% of the guinea-pigs. Ultradian VCO2 rhythms, with periods which show statistically significant interspecies differences, were assessed. For each of the 5 species these computed periods, which were the same in LL and DD, were: 1.17 h for quail and chickens, 1.25 h for rats, 1.50 h for mice and 1.0 h for guinea-pigs. In LD 12:12 these periods were different during L and D in quail, chicks, rats and mice, but not in guinea-pigs. The amplitudes of these ultradian variations were, according to the species, 10-20% of their mean VCO2 levels. These ultradian rhythms persist in the absence (or masking) of circadian rhythms, e.g. in LD 12:12 in 20% of guinea-pigs and in LL in 87% of Japanese quail and in 23% of Sprague-Dawley rats. Moreover, these ultradian rhythms persist during starvation, locomotor activity restraint and ageing. These ultradian VCO2 cycles which are related to rest-activity variations appear to be basic physiological rhythms with a genetic origin.     

Rhythmic cortisol secretion in the equine: Analysis and physiological mechanisms

Abstract

Four Thoroughbred mares (no. 1–4) were maintained under constant temperature (24°C) and controlled light (L/D:12/12 with lights on at 06.00 hr) conditions. They were fed and watered ad libitum with fresh feed and water given at 09.00 hr. After a 45‐day pre‐conditioning period, blood samples were obtained by veinipuncture at 4‐hr intervals for 14 days to determine circadian and day‐to‐day variation. The horses exhibited a circadian rhythm with maximum values attained at about 12.00 hr, however, there are periods of days in which no rhythm is distinguishable. Ultradian rhythms with mean periods of 105 to 128 and 24 to 31 min are superimposed upon the circadian rhythm. The individual rhythms are quite variable from horse to horse and within the same horse. During periods of decline in plasma cortisol with metabolic half‐lives of approximately 70 min, secretion of cortisol was very low or had ceased. During periods of increasing plasma concentration, secretion was occurring at a faster rate than degradation. Rapid decreases in plasma concentration (metabolic half‐life of approximately 30 min) was accompanied by a rise in specific activity indicating cortisol with a high specific activity was entering the plasma pool from other storage pools.     

Digestive Tract Cell Proliferation and Food Consumption Patterns of Ha/Icr Mice

The relationship between the daily pattern of food consumption and the proliferation rate of the qesophagus, stomach, forestomach, small intestine and colon of Ha/ICR mice was examined. Proliferative activity was determined by [³H]TdR incorporation on a wet weight tissue basis, along with selective counting of labelled nuclei. Under conditions of ad libitum feeding with a 12 hr light cycle (lights on at 0600) mice eat most of their food during the dark period. A distinct circadian rhythm was observed in the oesophagus, stomach, forestomach and colon with the peak of [³H]TdR incorporation between 0400 and 0600 and the nadir between 1600 and 1800. Although a circadian fluctuation was observed in the small intestine, its amplitude was much less than in other areas. This rhythmic change in proliferation rate could be phase shifted by allowing the mice to feed only between 0800 and 1600 for 14 days. Under these conditions the peak in proliferative activity occurred between 1800 and 2000. Fasting reduced the daily level of proliferative activity in all of the digestive tract sites studied, and for all areas except the oesophagus greatly reduced or eliminated the circadian fluctuation. the forestomach and colon were the most influenced by fasting with 24 hr [³H]TdR incorporation reduced to 30–40% of the control value. Refeeding following a 48 hr fast produced a rapid increase in proliferative activity peaking at levels well above the control value at 16 hr after the onset of refeeding. the major exception to this was the small intestine which slowly returned to the control value during the first 24 hr. Partial refeeding produced a diminished refeeding response. Once the normal pattern of food consumption was re-established following refeeding the normal proliferative fluctuations were again observed.     

Zonal expression of the glucokinase gene in rat liver. Dynamics during the daily feeding rhythm and starvation-refeeding cycle demonstrated by in situ hybridization

The abundance and zonal distribution of glucokinase (GK) mRNA were studied in rat liver during a normal 12 h day/12 h night rhythm (dark from 1900 to 0700 hours) and during refeeding after 60 h of starvation. Zonation of GK gene expression was examined by in situ hybridization with a radiolabelled cRNA probe and GK mRNA abundance was determined by Northern blot analysis with a digoxigenin-labelled cRNA probe. GK mRNA appeared to be almost homogeneously distributed throughout the whole daily feeding cycle; yet it was predominantly localized in the perivenous and intermediate zone during refeeding after 60 h of starvation. During the daily feeding rhythm, the total amount of GK mRNA increased quickly with the beginning of the feeding period at 1900 hours reaching a maximum at midnight and then decreased continuously to a basal level at noon. Virtually no GK mRNA was detected after 60 h of starvation. Refeeding caused a rapid increase in GK mRNA to a maximum at 2400 hours followed by a decrease to approximately two-thirds of the maximum value at 0700 hours. If the homogeneous distribution of GK mRNA during the daily feeding rhythm was real rather than apparent because of too low a sensitivity of the cRNA probe, the present results suggest that during the normal circadian cycle the mainly perivenous distribution of GK enzyme activity and protein is regulated preferentially at a translational level. The findings clearly show that during refeeding after 60 h of starvation the GK distribution is controlled predominantly at a pretranslational level.     

Lack of circadian rhythmicity of rat fetal intestinal enzymes as compared to the dams

The 24-hr activity patterns of intestinal maltase, lactase, leucylnaphthylamine hydrolyzing activity, γ-glutamyltransferase, and alkaline phosphatase were determined in pregnant rats maintained on a 12-12 light-dark cycle, with feeding during the dark period (1800-0600 hr, EST). The activities of these enzymes plus those of lysosomal maltase and lactase were followed during the same time period in 19- to 20-day-old fetuses. The activity patterns in the dams followed circadian rhythms, with peak activities occurring during the feeding-dark period. These rhythms are similar to the feeding schedule-cued rhythms observed in male rats and, therefore, are assumed to be feeding schedule cued also. In the fetuses, which obtained nutrients through the placenta, the activities increased in a somewhat nonlinear manner throughout the entire 24-hr period, but did not display a defined rhythm. It is concluded that endogenous intestinal enzyme rhythms do not exist in utero, and that oral and/or intermittent feeding is necessary for these rhythms to occur.     

The effect of various photoperiods on daily oscillations of serum corticosterone and insulin in rats.

The effect of various photoperiods on circadian rhythms of chosen parameters was investigated in laboratory rats. SPF male Wistar rats were adapted for six weeks to artificial light-dark cycles (LD 8:16, 12:12, 16:8). The light was switched on at 07.00 h in all regimens. The rats were killed at 3-hour intervals within 24 h, the serum concentration of corticosterone, insulin, glucose, food and water intake was determined. The external and computative acrophases of corticosterone varied in every photoperiod being dependent on the duration of light, the mesor values decreased in LD 16:8 in comparison with other photoperiods. The external acrophase of insulin was located 4 h after light onset in LD 8:16 and 12:12, in LD 16:8 one hour before light onset. The mesor values were approximately equal in all photoperiods. The circadian rhythms of glucose were similar in all regimens. Circadian variation of food and water consumption culminated at the same time in all regimens, the amount of food consumed in light increased with the light duration. Various photoperiods remarkably influenced circadian oscillations of corticosterone and in part food and water intake which could be considered as photoperiodic traits.     

Effects of Insulin Administration in Diabetic Rats on Food and Drink Rhythmic Schedules

In normal rats food and water intakes are associated in terms of time and quantity and their diurnal rhythms are synchronized. Intake behavior in streptozotocin-induced diabetic rats (ID) with marked polyphagia and polydipsia and in diabetic rats with continuous insulin administration (IT) has been studied. The daily percentages of food and water intakes during the dark phase were lower in IT than in control rats (C), being even lower in ID rats. However, all three groups showed circadian rhythmicity in food intake, although with less amplitude in the ID and IT animals compared to the C ones. A loss of the normal circadian rhythm of water intake was observed in the ID rats and although the insulin administration recovered circadian rhythmicity, it did not restore the temporal relations between food and water intakes. These results may indicate that the circadian pattern of water intake is more influenced by insulin than food intake. The daily pattern of this hormone may play an important role in the circadian modulation of the homeostatic mechanisms integrating both intake behaviors.     

Circadian rhythms of food and 1% NaCl intake, urine and electrolyte excretion, plasma renin activity and insulin concentration in adrenalectomized rats

The circadian rhythms of food and 1% NaCl intake, and urine, Na+, Cl- and K+ excretion were followed up in male Wistar rats before and one week after bilateral adrenalectomy at 4-hour intervals during two consecutive days. The circadian rhythms of plasma renin activity (PRA) and plasma immunoreactive insulin (IRI) were evaluated after decapitation of both intact and adrenalectomized rats at 08, 16 and 24 h. To all rats 1% NaCl was offered instead of drinking water. Adrenalectomy did not cause any significant phase shift in the cosine curves approximating the data collected at 4-hour intervals. The circadian rhythms showed the same relationships before and after the operation: the rhythms of food intake, K+ excretion and saline intake preceded significantly the rhythms of urine, Na+ and Cl- excretion. Adrenalectomy induced an increase in mean PRA and shifted its minimal value from 08 to 24 h. After the operation mean IRI decreased and the minimal value shifted from 16 to 24 h. It was concluded that adrenal glands do not play an important role in the synchronization of the circadian rhythms of food and 1% NaCl intake, urine and synchronization of the circadian rhythms of food and 1% NaCl intake, urine and electrolyte excretion with the illumination cycle, but play a relevant role in the synchronization of the circadian rhythms of PRA and IRI in the rat.     

Urinary excretion of immunoreactive prostaglandin E: A circadian rhythm and the effect of posture

The excretion of urinary immunoreactive prostaglandin E (iPGE), sodium, potassium, creatinine and volume was studied in 4 hr collections in normal women at normal activity. iPGE exhibited a circadian rhythm with an amplitude of 29% and peak excretion at 4:55 P.M. There were also significant circadian rhythms for sodium, potassium, creatinine, and volume, all peaking in late afternoon. There were no significant changes either in the total excretion or in the circadian rhythms of iPGE, potassium, or creatinine excretion when the subjects remained in bed for an entire day while the circadian rhythms of sodium and volume were significantly modified in amplitude and phase, respectively. Urinary aldosterone excretion decreased significantly when the subjects were at bed rest. iPGE excretion increased 33% when subjects were first recumbent and then erect for consecutive 4 hr periods on the same day (but when subjects were erect 1 day for a 4 hr period, iPGE excretion was lower by 32% than for the same 4 hr period the preceding day when they were recumbent). These data indicate that: 1) the sympathetic nervous system and renin-angiotensin-aldosterone system do not affect the circadian rhythm of urinary iPGE, and 2) short-term experiments of prostaglandin E excretion must be designed to avoid misleading results due to the circadian rhythm.     

Effects of excessive copper intake on hematological and hemorheological parameters

Copper plays an important role in the structure and function of metalloproteins and in the absorption of iron. The present study deals with the effects of excessive copper intake on hematological and hemorheological parameters. Drinking water containing 250 μg/mL copper for a period of 9 wk, Wistar albino rats showed increased erythrocyte count, blood viscosity, and hematocrit values (p<0.05) and lower hemoglobin (p<0.05) than controls fed a normal diet. The two groups also had differences in the erythrocyte deformability index. The results suggest that excessive copper intake results in hematological and hemorheological changes affecting both the protein content of the erythrocyte membrane and heme synthesis.     

Locomotory and stridulatory circadian rhythms in the cricket, Teleogryllus commodus

Male crickets of the species Teleogryllus commodus express circadian rhythms in both their stridulatory and locomotory behaviours. Both forms of activity show the same free-running period (τ) in either DD (23·4 hr) or LL (25·1 hr). Although some overlap is seen between periods of locomotion and stridulation, the majority of each activity is found in different phases of the circadian cycle: locomotion occurs mainly in the subjective day and stridulation in the subjective night. Entraining LD cycles with photoperiods of 12 hr produce exogenous effects that can obscure endogenous components of the rhythms. Red light (λ>600 nm) causes the period to lengthen and RD cycles can entrain both rhythms. Single white light pulses of 2 or 6 hr did not produce significant phase shifts, but did cause τ to shorten when given in the subjective night. The significance of these observations is discussed. Given the results obtained to date, it is not likely that each rhythm is under the control of a separate circadian pacemaker.     

Effects of constant darkness and constant light on circadian organization and reproductive responses in the ram

The relationship between circadian rhythms in the blood plasma concentrations of melatonin and rhythms in locomotor activity was studied in adult male sheep (Soay rams) exposed to 16-week periods of short days (8 hr of light and 16 hr of darkness; LD 8:16) or long days (LD 16:8) followed by 16-week periods of constant darkness (dim red light; DD) or constant light (LL). Under both LD 8:16 and LD 16:8, there was a clearly defined 24-hr rhythm in plasma concentrations of melatonin, with high levels throughout the dark phase. Periodogram analysis revealed a 24-hr rhythm in locomotor activity under LD 8:16 and LD 16:8. The main bouts of activity occurred during the light phase. A change from LD 8:16 to LD 16:8 resulted in a decrease in the duration of elevated melatonin secretion (melatonin peak) and an increase in the duration of activity corresponding to the changes in the ratio of light to darkness. In all rams, a significant circadian rhythm of activity persisted over the first 2 weeks following transfer from an entraining photoperiod to DD, with a mean period of 23.77 hr. However, the activity rhythms subsequently became disorganized, as did the 24-hr melatonin rhythms. The introduction of a 1-hr light pulse every 24 hr (LD 1:23) for 2 weeks after 8 weeks under DD reinduced a rhythm in both melatonin secretion and activity: the end of the 1-hr light period acted as the dusk signal, producing a normal temporal association of the two rhythms. Under LL, the 24-hr melatonin rhythms were disrupted, though several rams still showed periods of elevated melatonin secretion. Significant activity rhythms were either absent or a weak component occurred with a period of 24 hr. The introduction of a 1-hr dark period every 24 hr for 2 weeks after 8 weeks under LL (LD 23:1) failed to induce or entrain rhythms in either of the parameters. The occurrence of 24-hr activity rhythm in some rams under LL may indicate nonphotoperiodic entrainment signals in our experimental facility. Reproductive responses to the changes in photoperiod were also monitored. After pretreatment with LD 8:16, the rams were sexually active; exposure to LD 16:8, DD, or LL resulted in a decline in all measures of reproductive function. The decline was slower under DD than LD 16:8 or LL.(ABSTRACT TRUNCATED AT 400 WORDS)     

Evidence for time-of-day dependent effect of neurotoxic dorsomedial hypothalamic lesions on food anticipatory circadian rhythms in rats

The dorsomedial hypothalamus (DMH) is a site of circadian clock gene and immediate early gene expression inducible by daytime restricted feeding schedules that entrain food anticipatory circadian rhythms in rats and mice. The role of the DMH in the expression of anticipatory rhythms has been evaluated using different lesion methods. Partial lesions created with the neurotoxin ibotenic acid (IBO) have been reported to attenuate food anticipatory rhythms, while complete lesions made with radiofrequency current leave anticipatory rhythms largely intact. We tested a hypothesis that the DMH and fibers of passage spared by IBO lesions play a time-of-day dependent role in the expression of food anticipatory rhythms. Rats received intra-DMH microinjections of IBO and activity and body temperature (T(b)) rhythms were recorded by telemetry during ad-lib food access, total food deprivation and scheduled feeding, with food provided for 4-h/day for 20 days in the middle of the light period and then for 20 days late in the dark period. During ad-lib food access, rats with DMH lesions exhibited a lower amplitude and mean level of light-dark entrained activity and T(b) rhythms. During the daytime feeding schedule, all rats exhibited food anticipatory activity and T(b) rhythms that persisted during 2 days without food in constant dark. In some rats with partial or total DMH ablation, the magnitude of the anticipatory rhythm was weak relative to most intact rats. When mealtime was shifted to the late night, the magnitude of the food anticipatory activity rhythms in these cases was restored to levels characteristic of intact rats. These results confirm that rats can anticipate scheduled daytime or nighttime meals without the DMH. Improved anticipation at night suggests a modulatory role for the DMH in the expression of food anticipatory activity rhythms during the daily light period, when nocturnal rodents normally sleep.