Circadian Drinking Rhythms Research Articles

Vasopressin-deficient suprachiasmatic nucleus grafts re-instate circadian rhythmicity in suprachiasmatic nucleus-lesioned arrhythmic rats

It was investigated whether grafts of the suprachiasmatic nucleus could re-instate circadian rhythmicity in the absence of its endogenous vasopressin production and whether the restored rhythm would have the long period length of the donor. Grafts of 17-days-old vasopressin-deficient homozygous Brattleboro rat fetuses, homotopically placed in arrhythmic suprachiasmatic nucleus-lesioned Wistar rats, re-instated circadian drinking rhythm within 20–50 days similar as seen for grafts of heterozygous control fetuses. Period length of the recovered rhythm revealed a similar difference (average 24.3 vs 23.8 h) as reported for the rhythm between the adult Brattleboro genotypes. In all transplants, also those of the two-third non-recovery rats, a surviving suprachiasmatic nucleus was visible as a vasoactive intestinal polypeptide-positive neuronal cell cluster, whereas heterozygous transplants also revealed the complementary vasopressinergic cell part. Explanation of the absence of recovery failed since no undisputable correlation emerged between recovery of rhythm and vasoactive intestinal polypeptide, vasopressin and/or somatostatin immunocytochemical characteristics of the suprachiasmatic nucleus of the transplant. Special focus on the somatostatinergic neurons revealed their presence only occasionally near or in between the vasoactive intestinal polypeptidergic and (in the case of heterozygous grafts) vasopressinergic cell cluster. However their aberrant cytoarchitectural position appeared not to have affected the possibility to restore drinking rhythm of the suprachiasmatic nucleus-lesioned arrhythmic rat. It was concluded that grafted Brattleboro fetal suprachiasmatic nucleus develop their intrinsic rhythm conform their genotype and that vasopressin is not a crucial component in the maintenance nor in the transfer of circadian activity of the biological clock for drinking activity. Vasopressin of the suprachiasmatic nucleus may instead serve modulation within the circadian system.

Circadian rhythmicity of vasopressin levels in the cerebrospinal fluid of suprachiasmatic nucleus-lesioned and -grafted rats.

Transplantation of the fetal suprachiasmatic nucleus (SCN) in arrhythmic SCN-lesioned rats can reinstate circadian drinking rhythms in 40% to 50% of the cases. In the current article, it was investigated whether the failure in the other rats could be due to the absence of a circadian rhythm in the grafted SCN, using a circadian vasopressin (VP) rhythm in the cerebrospinal fluid (CSF) as the indicator for a rhythmic SCN. CSF was sampled in continuous darkness from-intact control rats and SCN-lesioned and -grafted rats. VP could be detected in all samples, with concentrations of 15 to 30 pg/ml in the control rats and 5 to 15 pg/ml in the grafted rats. A circadian VP rhythm with a two- to threefold difference between peak and nadir values was found in all 7 control rats but in only 4 of 13 experimental rats, despite the presence of a VP-positive SCN in all grafts. A circadian VP rhythm was present in 2 drinking rhythm-recovered rats (6 of 13) and in 2 nonrecovery rats. Apparently, in these latter rats, the failure of the grafted SCN to restore a circadian drinking rhythm cannot be attributed to a lack of rhythmicity in the SCN itself. Thus, the presence of a rhythmic grafted SCN, as is deduced from a circadian CSF VP rhythm, appears not to be sufficient for restoration of a circadian drinking rhythm in SCN-lesioned arrhythmic rats.

Circadian drinking during ad libitum and restricted feeding in pygmy goats

Eating and drinking of eight pygmy goats were recorded during ad libitum and restricted feeding conditions. During ad libitum conditions, water and food consumption occurred throughout the day with a circadian distribution of food and water intake showing peaks at the beginning of the light phase and at the beginning of the dark phase. When access to food was allowed only at the beginning (09:00–11:00 h) and at the end (19:00–21:00 h) of the 12 h light phase, a peak of water intake occurred during the two hours preceding and the first hour following dark onset. Therefore water/food ratio was greater during the evening hours (19:00–21:00 h) than during the morning hours (09:00–11:00 h). The results show that pygmy goats having access to food twice a day for 2 h at the beginning and towards the end of the 12 h light phase concentrate their water intake mainly during the last two hours before and the first hour after dark onset. This may contribute to the concomitant high food intake because there is evidence for control of feeding by the osmolarity of the ruminal fluid. Furthermore, the divergent water/food ratio between morning and evening hours suggest a dissociation of the circadian drinking rhythm from eating.

Neonatal clomipramine treatment, alcohol intake and circadian rhythms in rats.

Neonatal exposure to antidepressant monoamine re-uptake inhibitors produces a wide variety of effects on the behavior and physiology of adult rats which are consistent with features of clinical depression. Since depressed patients show characteristic alterations in circadian rhythmicity, our laboratory has examined free-running circadian drinking rhythms in this putative animal depression model. Previously, neonatal desipramine treatment was shown to lengthen free-running period, and increase circadian amplitude, spectral magnitude, and voluntary alcohol intake (10% ethanol v/v) of male rats. The purpose of the present study was to examine the effects of neonatal clomipramine treatment (25 or 30 mg/kg s.c., postnatal days 8-21) on circadian drinking rhythms and alcohol intake of both male and female rats. In addition, effects of alcohol exposure on circadian rhythmicity were also examined. Contrary to expectations, free-running period of clomipramine-treated rats did not differ from saline-treated controls in either constant darkness (DD) or constant light (LL), but spectral magnitude was increased in clomipramine-treated males and females, and circadian amplitude was increased in clomipramine-treated females. Neonatal clomipramine also increased voluntary alcohol intake, and both clomipramine- and saline-treated groups displayed significant period-shortening during alcohol exposure. Taken together, these results suggest that alterations in the amplitude and coherence of circadian rhythmicity may be more consistent than alterations in free-running period in animal depression models, as has been suggested previously for depressed patients.

Cellular Requirements of Suprachiasmatic Nucleus Transplants for Restoration of Orcadian Rhythm

Fetal neurografts containing the suprachiasmatic nucleus (SCN) can restore the circadian locomotor and drinking rhythm of SCN-lesioned (SCNX) rat and hamster. This functional outcome finally proves that the endogenous biological clock autonomously resides in the SCN. Observations on the cellular requirements of the “new” SCN for restoration of the arrhythmic SCNX animals have led to some new insights and confirmed findings from other studies. A critical mass of SCN neurons appeared necessary for functional effects, whereas the temporal profile of reinstatement of rhythm correlated with the delayed maturation of the grafted SCN. Cytoarchitectoni-cally, the grafted SCN does not seem to develop normally for all anatomical aspects. Complementary clusters of vasoactive intestinal polypeptide(VIP)-and vasopressin(VP)ergic neurons are formed, but somatostatin(SOM)ergic neurons do not always “join” this group, as is normally seen in situ. Nevertheless, these new SCNs can restore the ablated functions. As the period length of restored rhythms tends to vary, it might be that the grafted SCN underwent an altered or impaired maturation that resulted in a different setting of its clock mechanism. A prominent role of VIPergic neurons seems indicated by their presence in all functional grafts, but, although they may be required, these cells do not appear to be a sufficient condition for restoration of rhythm. Many grafts exhibit the presence of VIPergic cells without counteracting the arrhythmia, whereas VP- and SOMergic SCN neurons are usually present as well. Findings with VP-deficient Brattleboro rat grafts indicated that VP is not the primary obligatory signal of circadian activity. It is argued that perhaps the role of SOMergic neurons in the clock function of the (grafted) SCN has been insufficiently considered. However, one should keep in mind that the peptides of the various types of SCN neurons may function only as cofactors, mutually modulating molecular or bioelectrical cellular activities within the nucleus or the message of the main transmitter γ-aminobutyric acid.

Effect of Ω‐conotoxin, a calcium channel blocker, on the circadian rhythm in rats

Abstract Using an Alzet osmotic minipump, we examined the effect of continuous infusion of O‐conotoxin, a N‐type voltage‐sensitive calcium channel blocker, into the third cerebral ventricle above the suprachiasmatic nucleus (SCN) of the hypothalamus on the circadian drinking rhythm of rats maintained under a 12‐h light and 12‐h dark cycle and constant darkness. O‐Conotoxin (10‐6 mol/h) infusion disrupted the rhythm in both conditions. In contrast, infusions of nifedipine (104 mol/h), a dihydropyridine sensitive L‐type calcium channel blocker, did not eliminate the rhythm. These findings suggest that N‐type voltage‐sensitive calcium channels are involved in the mechanism of generation of the circadian rhythm driven by the circadian oscillator in the SCN.

Neonatal desipramine treatment alters free-running circadian drinking rhythms in rats

Neonatal treatment with monoamine reuptake inhibitors results in a constellation of neurobehavioral alterations in adult rats that may model human depression. Since alterations in circadian rhythmicity have been reported in both depressed patients and in animal depression models, the present study examined the effects of neonatal desipramine treatment (5.0 mg/kg SC from postnatal day 7 through 22) on free-running circadian drinking rhythms. Rhythmicity was examined in constant darkness (DD), constant light (LL), and during adult desipramine treatment (0.25 mg/ml via the drinking water). Compared with saline-treated controls, neonatal desipramine lengthened free-running period in DD, blunted the period-altering effect of LL, and potentiated the period-altering effect of adult desipramine treatment. Neonatal desipramine treatment also increased circadian amplitude and spectral magnitude, but did not modify the effects of light or adult desipramine on these parameters. These results provide further evidence that behavioral depression is associated with alterations in circadian rhythmicity, and are consistent with the hypothesis that such relationships are mediated by brain monoaminergic systems.

Circadian drinking rhythms in SHR and WKY rats: Effects of increasing light intensity

This study sought to define the generality of a previous finding that the spontaneously hypertensive (SHR) and normotensive Wistar-Kyoto (WKY) rat strains differ in free-running circadian period when maintained in running-wheel cages under constant light. Circadian drinking rhythms were monitored in SHRs and WKYs housed without access to running wheels under an increasing series of light intensities beginning with constant darkness. Strain differences in circadian period were seen only at relatively high light intensities, indicating that SHRs and WKYs differ in circadian light sensitivity. Since SHRs and WKYs differ in circadian period with or without access to running wheels, this strain difference is not likely to depend on differential locomotor activity levels. SHRs and WKYs also differed in spectral profile and circadian waveform, but only under low light intensities. At higher intensities, dissociation of rhythmicity was seen in both strains.

Functional development of fetal suprachiasmatic nucleus grafts in suprachiasmatic nucleus-lesioned rats

Recovery of arcadian drinking rhythms in Suprachiasmatic nucleus (SCN)-lesioned rats after fetal SCN grafting was related to the immunocytochemical appearance and fiber outgrowth of vasopressin (VP)-, vasoactive intestinal polypeptide (VIP)-, and somatostatin (SOM)-containing neurons in the implants. At 4 weeks postgrafting, the first recovered animal was found. After longer survival times, 38% of the animals showed recovery. Immunocytochemical evaluation indicated that full maturation of the SCN grafts was not reached until 4 weeks postgrafting. Grafted VP and VIP cells were always located together, whereas SOM cells were clustered nearby but separate. Neuropeptide Y fibers were observed with an increasing fiber density between 2 and 5 weeks posttransplantation and were clustered particularly at the level of the SOM cells. In all rhythm-recovered animals transplants of VP and VIP fibers had grown laterally into the hypothalamus. A few nonrecovered animals also showed ingrowth of such fibers, though more caudally to the lesioned SCN. Many of the nonrecovered rats showed similar stainings but without these efferent outgrowth to the host. We conclude that neither a humoral factor nor the presence of VP and VIP efferents in the host brain alone are enough for the restoration of circadian drinking rhythms.

Further evaluation of the tetrodotoxin-resistant circadian pacemaker in the suprachiasmatic nuclei.

We previously reported the results of an experimental paradigm in which tetrodotoxin (TTX) was chronically infused by miniosmotic pump into the rat suprachiasmatic nuclei (SCN) (Schwartz et al., 1987). Although TTX reversibly blocked photic entrainment and overt expression of the circadian drinking rhythm, the circadian pacemaker in the SCN continued to oscillate unperturbed by the toxin, and we concluded that Na(+)-dependent action potentials are not a part of the SCN pacemaker's internal timekeeping mechanism. In the research reported in the present paper, we used our paradigm to chronically infuse other agents, in order to evaluate the validity of this interpretation further. (1) Infusion of 50% procaine into the SCN of blinded rats resulted in a disorganized circadian drinking rhythm during the infusion, after which behavioral rhythmicity returned without apparent phase shift. In intact rats, procaine reduced the phase-resetting action of a reversed light-dark cycle imposed during the infusion. Thus, the effects of voltage-dependent Na+ channel blockade by a local anesthetic resemble those produced by TTX. (2) Infusion of high (20 mM) K+ or 100 microM veratridine into the SCN of blinded rats resulted in an apparent phase advance of the circadian drinking rhythm by over 4 hr. The phase-shifting effect of veratridine was blocked by simultaneous infusion of 1 microM TTX. Thus, membrane depolarization or direct activation of voltage-dependent Na+ channels can affect the pacemaker's oscillation. Our infusion paradigm can detect alterations of rhythm phase, and the lack of phase shift after TTX or procaine infusion is not an artifact of an insensitive method.

The effects of feedback lighting on the circadian drinking rhythm in the diurnal new world primate Saimiri sciureus.

Feedback lighting provides illumination primarily during the subjective night (i.e., the photosensitive portion of the circadian cycle) in response to a given behavior. This technique has previously been used to test the nonparametric model of entrainment in nocturnal rodents. In three species (Rattus norvegicus, Mesocricetus auratus, and Mus musculus), the free-running period of the locomotor activity rhythm was similar whether the animals were exposed to continuous light or discrete light pulses occurring essentially only during the subjective night (i.e., feedback lighting). In the current experiments, feedback lighting was presented to squirrel monkeys so that light fell predominantly during the subjective night. Feedback lighting was linked to the drinking behavior in this diurnal primate so that when the animal drank, the lights went out. Despite the seemingly adverse predicament, the monkeys maintained regular circadian drinking rhythms. Furthermore, just as the period of the free-running activity rhythms of nocturnal rodents exposed to continuous light or feedback lighting were similar, the period of the drinking rhythms of the squirrel monkeys in continuous light and feedback lighting were comparable (25.6 +/- 0.1 and 25.9 +/- 0.1 hours, respectively), despite a substantial decrease in the total amount of light exposure associated with feedback lighting. The free-running period of monkeys exposed to continuous dark (24.5 +/- 0.1 hours) was significantly shorter than either of the two lighting conditions (P < 0.001). The results presented for the drinking rhythm were confirmed by examination of the temperature and activity rhythms. Therefore, discrete light pulses given predominately during the subjective night are capable of simulating the effects of continuous light on the free-running period of the circadian rhythms of a diurnal primate. The response of squirrel monkeys to feedback lighting thus lends further support for the model and suggests that the major entrainment mechanisms are similar in nocturnal rodents and diurnal primates.

The suprachiasmatic nuclei contain a tetrodotoxin-resistant circadian pacemaker.

Tetrodotoxin was infused into the suprachiasmatic nuclei of unanesthetized and unrestrained rats continuously for 14 days. The internal timekeeping mechanism of the circadian pacemaker in the nuclei continued to oscillate unaffected by this treatment, although the toxin reversibly blocked function of both the input pathway for pacemaker entrainment and an output pathway for expression of the circadian drinking rhythm. Thus, Na+-dependent action potentials appear necessary for entrainment and expression of overt circadian rhythms, but they do not seem necessary for the pacemaker to keep accurate time. The experimental approach presented in this paper is useful because it allows systematic assessment and distinction of the input, pacemaker, and output components of a mammalian circadian timekeeping system in vivo.

Circadian drinking rhythms and blood alcohol levels in two rat lines developed for their alcohol consumption

The circadian drinking behavior of the AA (Alko Alcohol) and ANA (Alko Non-Alcohol) rat lines developed for high and low voluntary alcohol consumption was studied by using a microcomputer lick-counting system. The blood alcohol concentrations of AA rats were measured during the drinking peaks. The results showed that AA rats voluntarily drink intoxicating amounts of alcohol. The highest blood alcohol levels were about 25 mmol/l. Differences in circadian drinking rhythms between AA and ANA lines were also found. AA rats had a trimodal drinking pattern for alcohol intake during the dark period but ANA rats drank alcohol rather evenly throughout the period.

Entrainment of circadian rhythms: Retinofugal pathways and unilateral suprachiasmatic nucleus lesions

The role of retinohypothalamic input to the suprachiasmatic nuclei (SCN) as well as SCN afferents from the ventrolateral geniculate nuclei (LGN v) in the entrainment of circadian drinking rhythms was investigated in the rat. Bilateral lesions of the LGN v and the primary optic tracts had no affect on the entrainment of drinking rhythms to a light-dark cycle, the response to a 12 hr phase shift of the light-dark cycle, or on the period of the free-running circadian rhythm in constant light or constant darkness. Unilateral blinding in rats with or without LGN v lesions retarded the rate of phase shifting by 2 days and decreased the period of the free-running rhythm in constant light. For rats with unilateral SCN lesions, or such lesions combined with either ipsi- or contra-lateral blinding, the rate of re-entrainment was intermediate between intact and unilaterally blinded rats indicating that unilateral SCN lesions partially reversed the effects of unilateral blinding. Unilateral SCN lesions had no effect on the period of the free-running rhythm in constant light or darkness. These results are consistent with the interpretation that the asymmetrical innervation of the two SCN by the RHT in unilaterally blinded rats delays re-entrainment by changing the phase response curve of the circadian system. This change may be mediated by neural connections between the two SCN.

Entrainment and masking of circadian drinking rhythms in primates: Influence of light intensity

The entrained drinking rhythms of squirrel monkeys were studied during exposure to 24 hr illumination cycles of three different intensities (60:0; 66:6; 76:16 lux). Increasing the intensity of ambient illumination significantly delayed the offset of drinking but had no effect on either the onset or the total amount of daily drinking behavior. Comparison of drinking behavior under an alternating schedule of LD 66:6 lux and constant light of 6 lux indicated that the twice daily light transitions consistently altered the temporal distribution of drinking behavior. The daily timing of squirrel monkey drinking behavior thus, depends not only on the mechanisms of circadian entrainment to the LD cycle, but also on the ability of the LD cycle to directly influence, or “mask” behavior.