Circadian Wheel-Running Activity of Rats Under Schedules of Limited Daily Access to Salt

Schedules of limited daily access to food result in 'anticipatory' activity preceding each daily feeding. It is well-established that such food-anticipatory activity depends on a food-entrainable circadian timing mechanism. In the present study, we sought to extend the generality of these results by maintaining rats in running wheels under schedules which provided access to salt solutions or to salty food for 2 hr each day. The animals were subjected to dietary, pharmacological and surgical treatments that promote salt appetite. However, we found no evidence for daily salt-anticipatory wheel running activity in any condition.     

Circadian Wheel-Running Activity of Rats Under Schedules of Limited Daily Access to Salt

Schedules of limited daily access to food result in 'anticipatory' activity preceding each daily feeding. It is well-established that such food-anticipatory activity depends on a food-entrainable circadian timing mechanism. In the present study, we sought to extend the generality of these results by maintaining rats in running wheels under schedules which provided access to salt solutions or to salty food for 2 hr each day. The animals were subjected to dietary, pharmacological and surgical treatments that promote salt appetite. However, we found no evidence for daily salt-anticipatory wheel running activity in any condition.     

Circadian Variations in the Number of Actively Secreting Sebaceous Follicles and Androgen Circadian Rhythms

Sebum excretion has been shown to demonstrate a circadian rhythm using a gravimetric method (cigarette paper). With the newly introduced method of Sebutape, we confirmed this periodicity and showed that the elevation in sebum excretion is correlated with an increase in the number of secreting follicles. We found, furthermore, that the number of secreting follicles on the forehead showed a distinct and statistically significant circadian rhythmicity, in contrast to those of the chest, which remained almost constant. The quantification in plasma levels of cortisol, melatonin, δ-4-androstenedione, dehydroepiandrosterone sulphate, and free testosterone showed no correlation with sebum excretion at either site. These observations suggest that local factors are involved in control of sebum secretion.     

Significance of Nonparametric Light Effects in Entrainment of Circadian Rhythms in Owl Monkeys (Aotus Lemurinus Griseimembra) by Light-Dark Cycles

In a total of 12 adult Colombian owl monkeys, Aotus lemurinus griseimembra, the significance of nonparametric light effects for the entrainment of the circadian system by light-dark (LD) cycles was studied by carrying out (a) phase-response experiments testing the phase-shifting effect of 30-min light pulses (LPs) of 250 lx applied at various phases of the free-running circadian activity rhythm (LL 0.2 lx) and (b) synchronization experiments testing the entraining effect of 24-h single LP photoperiods consisting of 30-min L of 80 lx and 23.5-h D of 0.5 lx (sP 0.5) and skeleton photoperiods consisting of two 30-min LPs of 80 lx, given against a background illuminance of 0.5 lx either symmetrically at 12-h intervals (PP 12:12) or asymmetrically at 9- and 15-h intervals (PP 9:15). The phase-response characteristics in Aotus, as evidenced by the phase-response curve, generally correspond to those of nocturnal rodents, proving that this neotropical simian primate chronobiologically is a genuine nocturnal species. When free-running with a spontaneous period close to 24 h (24.3 ± 0.1 h), the PP 12:12 produced entrainment in only two of five owl monkeys, whereas the sP 0.5 entrained four of them. The PP 9:15, however, brought about stable entrainment of the circadian rhythms of locomotor activity, feeding activity, and core temperature in all animals tested (n = 8). Changes in phase position of the activity time with the endogenous rhythm entrained by a PP 12:12, by an sP 0.5, or by a PP 9:15 give evidence that both LPs of a skeleton photoperiod contribute to the phase setting of the circadian system. When free-running with a considerably lengthened spontaneous period (τ ≥ 25.5 h), even the sP 0.5 and the PP 9:15 failed to entrain the owl monkeys' circadian rhythms, whereas a 24-h photoperiod with a very long LP of 3 h caused entrainment. The results indicate that in Aotus lemurinus griseimembra, in addition to the nonparametric light effects, parametric light effects play a significant role in the entrainment of circadian rhythms by LD cycles.     

Circadian Rhythms of Locomotor Activity in the Nile Tilapia Oreochromis niloticus

Behavioral rhythms of the Nile tilapia were investigated to better characterize its circadian system. To do so, the locomotor activity patterns of both male and female tilapia reared under a 12:12 h light-dark (LD) cycle were studied, as well as in males the existence of endogenous rhythmicity under free-running conditions (DD and 45 min LD pulses). When exposed to an LD cycle, the daily pattern of activity differed between individuals: some fish were diurnal, some nocturnal, and a few displayed an arrhythmic pattern. This variability would be typical of the plastic circadian system of fish. Moreover, reproductive events clearly affected the behavioral rhythms of female tilapia, a mouth-brooder teleost species. Under DD, 50% (6 of 12) of male fish showed circadian rhythms with an average period (τ) of 24.1±0.2 h, whereas under the 45 min LD pulses, 58% (7 of 12) of the fish exhibited free-running activity rhythms with an average τ of 23.9±0.5 h. However, interestingly in this case, activity was always confined to the dark phase. Furthermore, when the LD cycle was reversed, a third of the fish showed gradual resynchronization to the new phase, taking 7–10 days to be completely re-entrained. Taken together, these results suggest the existence of an endogenous circadian oscillator that controls the expression of locomotor activity rhythms in the Nile tilapia, although its anatomical localization remains unknown.     

Effects of Pregnancy and Parturition on Free-Running Circadian Activity Rhythms in the Rat

Alterations in circadian rhythms have previously been associated with estrous and seasonal changes in reproductive state. In the present study we explored the effects of the reproductive events of pregnancy and parturition on free-running circadian activity rhythms in the rat. Free-running rhythms were monitored before mating, during pregnancy, and following parturition and removal of pups. Systematic and long-lasting alterations of the period of the free-running activity rhythm were seen following parturition. The effects of estrous, seasonal, and gestational reproductive states on circadian rhythms may be mediated by the endocrine events which accompany these states.     

The use of Constant Routines in Unmasking the Endogenous Component of Human Circadian Rhythms

A major problem in the study of the internal clock(s) that drives human circadian rhythms is that due to the effect produced by rhythmicity of habits and external influences (‘masking’). A particularly potent factor in this respect is the sleep-wake cycle. It is anomalous that, even though this masking influence is widely accepted, most studies of circadian rhythmicity have been performed in the presence of such interferences.

A protocol is described, the constant routine, by which these exogenous influences can be minimized, thereby enabling a closer scrutiny of the internal clock(s) to be made. An account is given of the different circumstances in which the constant routines have been used together with the results derived from such studies. Briefly, they indicate that nychthemeral studies can give misleading information about the rate of adjustment of the internal clock to various manipulations, e.g. time-zone transition, shift work.

In addition, future studies making use of constant routines are described, in particular those which might enable the presence of more than one internal clock to be established.     

Circadian Discrimination of Reward: Evidence for Simultaneous Yet Separable Food- and Drug-Entrained Rhythms in the Rat

A unique extra-suprachiasmatic nucleus (SCN) oscillator, operating independently of the light-entrainable oscillator, has been hypothesized to generate feeding and drug-related rhythms. To test the validity of this hypothesis, sham-lesioned (Sham) and SCN-lesioned (SCNx) rats were housed in constant dim-red illumination (LL_red) and received a daily cocaine injection every 24?h for 7 d (Experiment 1). In a second experiment, rats underwent 3-h daily restricted feeding (RF) followed 12 d later by the addition of daily cocaine injections given every 25?h in combination with RF until the two schedules were in antiphase. In both experiments, body temperature and total activity were monitored continuously. Results from Experiment 1 revealed that cocaine, but not saline, injections produced anticipatory increases in temperature and activity in SCNx and Sham rats. Following withdrawal from cocaine, free-running temperature rhythms persisted for 2–10 d in SCNx rats. In Experiment 2, robust anticipatory increases in temperature and activity were associated with RF and cocaine injections; however, the feeding periodicity (23.9?h) predominated over the cocaine periodicity. During drug withdrawal, the authors observed two free-running rhythms of temperature and activity that persisted for >14 d in both Sham and SCNx rats. The periods of the free-running rhythms were similar to the feeding entrainment (period?=?23.7 and 24.0?h, respectively) and drug entrainment (period?=?25.7 and 26.1?h, respectively). Also during withdrawal, the normally close correlation between activity and temperature was greatly disrupted in Sham and SCNx rats. Taken together, these results do not support the existence of a single oscillator mediating the rewarding properties of both food and cocaine. Rather, they suggest that these two highly rewarding behaviors can be temporally isolated, especially during drug withdrawal. Under stable dual-entrainment conditions, food reward appears to exhibit a slightly greater circadian influence than drug reward. The ability to generate free-running temperature rhythms of different frequencies following combined food and drug exposures could reflect a state of internal desynchrony that may contribute to the addiction process and drug relapse. (Author correspondence: heiko@vetmed.wsu.edu)     

Scheduled Feeding Alters the Timing of the Suprachiasmatic Nucleus Circadian Clock in Dexras1-Deficient Mice

Restricted feeding (RF) schedules are potent zeitgebers capable of entraining metabolic and hormonal rhythms in peripheral oscillators in anticipation of food. Behaviorally, this manifests in the form of food anticipatory activity (FAA) in the hours preceding food availability. Circadian rhythms of FAA are thought to be controlled by a food-entrainable oscillator (FEO) outside of the suprachiasmatic nucleus (SCN), the central circadian pacemaker in mammals. Although evidence suggests that the FEO and the SCN are capable of interacting functionally under RF conditions, the genetic basis of these interactions remains to be defined. In this study, using dexras1-deficient (dexras1^?/?) mice, the authors examined whether Dexras1, a modulator of multiple inputs to the SCN, plays a role in regulating the effects of RF on activity rhythms and gene expression in the SCN. Daytime RF under 12L:12D or constant darkness (DD) resulted in potentiated (but less stable) FAA expression in dexras1^?/? mice compared with wild-type (WT) controls. Under these conditions, the magnitude and phase of the SCN-driven activity component were greatly perturbed in the mutants. Restoration to ad libitum (AL) feeding revealed a stable phase displacement of the SCN-driven activity component of dexras1^?/? mice by ～2?h in advance of the expected time. RF in the late night/early morning induced a long-lasting increase in the period of the SCN-driven activity component in the mutants but not the WT. At the molecular level, daytime RF advanced the rhythm of PER1, PER2, and pERK expression in the mutant SCN without having any effect in the WT. Collectively, these results indicate that the absence of Dexras1 sensitizes the SCN to perturbations resulting from restricted feeding. (Author correspondence: haiying.cheng@utoronto.ca)     

Effect of Sex,Menstrual Cycle Phase,and Oral Contraceptive Use on Circadian Temperature Rhythms

The circadian rhythm of rectal temperature was continuously recorded over several consecutive days in young men and women on regular nocturnal sleep schedules. There were 50 men, 21 women with natural menstrual cycles [i.e., not taking oral contraceptives (OCs) (10 in the follicular phase and 11 in the luteal phase)], and 14 women using OCs (6 in the pseudofollicular phase and 8 in the pseudoluteal phase). Circadian phase and amplitude were estimated using a curve-fitting procedure, and temperature levels were determined from the raw data. A two-way analysis of variance (ANOVA) on the data from the four groups of women, with factors menstrual cycle phase (follicular, luteal) and OC use (yes, no), showed that temperature during sleep was lower during the follicular phase than during the luteal phase. Since waking temperatures were similar in the two phases, the circadian amplitude was also larger during the follicular phase. The lower follicular phase sleep temperature also resulted in a lower 24-h temperature during the follicular phase. The two-way ANOVA showed that temperature during sleep and 24-h temperature were lower in naturally cycling women than in women taking OCs. A one-way ANOVA on the temperature rhythm parameters from the five groups of subjects showed that the temperature rhythms of the men and of the naturally cycling women in the follicular phase were not significantly different. Both of these groups had lower temperatures during sleep, lower 24-h temperatures, and larger circadian amplitudes than the other groups. There were no significant differences in circadian phase among the five groups studied. In conclusion, menstrual cycle phase, OC use, and sex affect the amplitude and level, but not the phase, of the overt circadian temperature rhythm.     

Review of the Dual-Clock Control of Tidal Rhythms and the Hypothesis that the Same Clock Governs Both Circatidal and Circadian Rhythms

Discoveries first published in 1986 did not fit the de rigueur working hypothesis that the clocks governing tide-associated rhythms had a fundamental period of 12.4 h, a value equal to the average interval between successive tides on most coastlines of the world. To explain the results a dual-clock schema was fashioned that envisioned two clocks, strongly coupled together 180° antiphase, each running at a basic rate of 24.8 h (the interval of a lunar day), as the driving agents of tide-associated rhythms (details are given in the text). This elaboration has been named the circalunidian-clock hypothesis, a hypocorism used in some armchair ruminations back in 1973. In the decade since 1986, a goodly amount of evidence has been garnered that is consistent with this hypothesis—suggesting that first-call divination appears to have been visionary. Acceptance of this hypothesis leads to further cerebration that a 24.8-h clock, its circa periods in constant conditions, and other properties—which fully overlap with our perception of the circadian clock that drives daily rhythms—may indicate that circadian and circalunidan timepieces are not different entities. The known properties of both daily and lunar clock-types are compared and contrasted, and, with the exception of one feature (for which there is at least a philosophical explanation), it is concluded that the same clock that drives tidal rhythms could also motor daily rhythms, i.e., there may be no such thing as a 12.4-h horologue.     

Investigating the Endogenous Component of Human Circadian Rhythms: A Review of Some Simple Alternatives to Constant Routines

Several types of constant routine are accepted as an important means by which the endogenous component of circadian rhythms can be studied. Nevertheless, they are impracticable to perform and unsuitable for routine use in many individuals. We describe a group of simple methods with which rhythms measured in normal circumstances can be dissociated into the components due to masking and the internal clock. Each method is best suited to a particular type of experimental condition. Results from a variety of protocols are analysed by these and conventional methods to assess the validity of the new methods.     

Ontogeny of the Circadian System During Embryogenesis in Rainbow Trout (Oncorhynchus mykyss) and the Effect of Prolonged Exposure to Continuous Illumination on Daily Rhythms of per1, clock,and aanat2 Expression

It is widely held that the development of the circadian system during embryogenesis is important for future survival of an organism. Work in teleosts has been, to date, limited to zebrafish, which provides little insight into the diversity of this system within such a large vertebrate class. In this study, the authors analyzed the diel expression of per1, clock, and aanat2 in unfertilized rainbow trout oocytes and embryos maintained under either a 12:12-h light:dark (LD) cycle or continuous illumination (LL) from fertilization. 24-h profiles in expression were measured at fertilization as well as 8, 21 42, and 57 days postfertilization (dpf). Both per1 and clock were expressed in unfertilized oocytes and all embryonic stages, whereas aanat2 expression was only measureable from 8 dpf. A reduction in both per1 and clock mean expression levels between unfertilized oocytes/0–1 dpf embryos and 8–9 dpf embryos was suggestive of a transition from maternal RNA to endogenous mRNA expression. Although aanat2 expression was not clearly associated with photic conditions, photoperiod treatment did alter the expression of per1 and clock expression/rhythmicity from as early as 8 dpf (per1), which could suggest the presence and functionality of an as yet unidentified “photoreceptor.” As a whole, this work demonstrates that clock systems are present and functional during embryonic development in rainbow trout. Further studies of their expression and regulation will help understand how the environment interacts with embryonic development in the species. (Author correspondence: andrew.davie@stir.ac.uk)     

Consistent Changes in the Circadian Rhythms of Blood Pressure and Atrial Natriuretic Peptide in Congestive Heart Failure

We demonstrated in previous works that the circadian rhythms of blood pressure (BP) and atrial natriuretic peptide (ANP) are antiphasic in normal subjects and in essential hypertension. The aim of the present study was to assess the circadian rhythms of BP and ANP in 20 patients with stable congestive heart failure (CHF), divided into two groups of 10 according to their New York Heart Association functional class. A matched control group of 10 normal volunteers was also studied. Noninvasive BP monitoring at 15-min intervals was performed for 24 h. Peripheral blood samples were also obtained at 4-h intervals starting from 08:OO h. The mean (±SEM) circadian mesors of ANP plasma levels were 13.4 ± 1.7 pmol/L in the control group, 28.6 ± 2.4 pmol/L in the group of 10 patients in class 11, and 81.5 ± 12 pmol/L in the group of 10 patients in class 111-IV. In normal subjects, plasma ANP concentration was highest at 04:OO h (21.5 ± 2.7 pmol/L) and lowest at 16:OO h (8.8 ± 2.4 pmol/L; p < 0.01). Both groups of patients with CHF showed no significant circadian change in the plasma levels of ANP and also a significantly blunted circadian rhythm of BP. Cosinor analysis confirmed the loss of the circadian rhythms of ANP and BP in CHF patients. Our findings support the existence of a causal relationship between the circadian rhythms of ANP and BP.     

Circadian and Ultradian (12 H) Rhythms of Hepatic Thiosulfate Sulfurtransferase (Rhodanese) Activity in Mice During the First Two Months of Life

Thiosulfate sulfurtransferase (TST) is an important ‘enzyme of protection,’ that accelerates the detoxification of cyanide, converting it into thiocyanate. The TST physiological rhythm was investigated at wks 2, 4, and 8 of post‐natal development (PND) in the mouse. The results revealed a statistically significant gender‐related difference, with the highest activity in females, at all the documented PND stages. In the second week of PND (pre‐weaning time), the circadian rhythm of the enzyme activity was associated with ultradian components. The prominent circadian rhythm (τ=24 h) peaked at the beginning of the light span, more precisely ～3 HALO (Hours After Light Onset). A week after weaning (wk 4 of PND), an impairment of the rhythm, with the peak shifted toward the second half of photophase, was recorded. Four to 6 wks later, about wk 8 of PND, the circadian rhythm pattern was stabilized, with its peak then located at the beginning of the dark span (13 HALO). The obtained results showed a 12 h phase‐shift of the circadian TST peak time during PND, suggesting that the rhythm stabilization is age‐dependent.     

Circadian Rhythms of Pollen and Nectar Collection by Bees on the Flowers of Ludwigia elegans (Onagraceae)

The visits of Tetraglossula ventralis (Hymenoptera: Colletidae) and Heterosarellus sp (Hymenoptera: Andrenidae) to collect pollen and nectar on the flowers of Ludwigia elegans (Onagraceae) were visually monitored from November 1989 to October 1991, in two localities of the State of São Paulo, Brazil. These localities are approximately at the same latitude (Mairinque 23° 52’ S and Campos do Jordão 22° 45’ S), but with a difference of 1000m in the altitude between them. As same latitudes ensure similar photoperiods and the difference in altitude, a persistent step in the temperature, the field work was conducted in environmental conditions varying within a known and limited range. Circadian rhythms are described for pollen and nectar harvesting behaviours, which are finely adjusted to the flower’s anthesis and withering. The environmental light/dark cycle is suggested as the main zeitgeber and temperature cycle either as a secondary zeitgeber or as a masking agent. Since these bee species are non-social, the possibility of social synchonization was discarded. We concluded that temporal adaptation plays a central role in the interaction of flowers and its visitors. In addition to specialized structures to collect the pollen of L.elegans, the acrophases of pollen and nectar collection rhythms coincide with the time when the stigma is most receptive and consequently the possibility of pollination increases highly.     

Circadian rhythmicity in the stimulation of bioluminescence by biogenic amines and MAO inhibitors inGonyaulax polyedra

In the dinoflagellateGonyaulax polyedra bioluminescence was investigated in constant darkness. Light emission was stimulated considerably and specifically by the biogenic smines epinephrine, 5-methoxytryptamine, and kynuramine. Various analogues and motabolites of these substances, such as norepinephrine, isoproterenol, phenylephrine, synephrine, metanephrine isoproterenol, phenylephrine, synephrine, metanephrine dopamine, 3,4-dihydroxymandelic and 3-methoxy hydroxymandelic acids, serotonin, N-acetylserotonin, melatonin, 5-hydroxytryptophol, 5-methoxytryptophol, kynurenine, 4-hydroxyquinoline, 3-hydroxyanthrani ic, and quinolinic acids were much less effective. Strong enhancement of bioluminescence, in the range of those obtained with the three stimulatory biogenic amines was also observed after administration of several compeunds acting as MAO inhibitors in mammalian systems, in particular, pargyline, amitriptyline,p-benzoquinone, tranylcypromine, harmaline, and noreleagnine. The responsiveness of cells towards epinephrine, 5-methoxytryptamine, kynuramine, amitriptyline,p-benzoquinone, and noreleagnine varied considerably within the circadian cycle, with the highest stimulations obtained during subjective night. These rhythms can be only partially explained by periodic bioluminescence capacity, and seem to comprise a cyclicity in the sensitivity of cells to the compounds mentioned.Paper presented at the 12th International Congress of Biometeorology (Vienna 1990)