Temporal reorganization of the suprachiasmatic nuclei in hamsters with split circadian rhythms.

A dual oscillator basis for mammalian circadian rhythms is suggested by the splitting of activity rhythms into two components in constant light and by the photoperiodic control of pineal melatonin secretion and phase-resetting effects of light. Because splitting and photoperiodism depend on incompatible environmental conditions, however, these literatures have remained distinct. The refinement of a procedure for splitting hamster rhythms in a 24-h light-dark:light-dark cycle has enabled the authors to assess the ability of each of two circadian oscillators to initiate melatonin secretion and to respond to light pulses with behavioral phase shifting and induction of Fos-immunoreactivity in the suprachiasmatic nuclei (SCN). Hamsters exposed to a regimen of afternoon novel wheel running (NWR) split their circadian rhythms into two distinct components, dividing their activity between the latter half of the night and the afternoon dark period previously associated with NWR. Plasma melatonin concentrations were elevated during both activity bouts of split hamsters but were not elevated during the afternoon period in unsplit controls. Light pulses delivered during either the nighttime or afternoon activity bout caused that activity component to phase-delay on subsequent days and induced robust expression of Fos-immunoreactivity in the SCN. Light pulses during intervening periods of locomotor inactivity were ineffective. The authors propose that NWR splits the circadian pacemaker into two distinct oscillatory components separated by approximately 180 degrees, with each expressing a short subjective night.     

Entrainment of 2 subjective nights by daily light:dark:light:dark cycles in 3 rodent species

Recent work with exotic 24-h light:dark:light:dark (LDLD) cycles indicates surprising flexibility in the entrainment patterns of Syrian hamsters. Following exposure to an LDLD cycle, hamsters may adopt a form of rhythm splitting in which markers of subjective night (e.g., activity, melatonin) are expressed in each of the twice daily scotophases. This pattern contrasts markedly with that of conventionally entrained hamsters in which markers of subjective night are expressed once daily in only 1 of the 2 dark periods. The "split" entrainment pattern was examined further here in Syrian and Siberian hamsters and in mice exposed to LDLD 7:5:7:5, a condition that reliably induces split activity rhythms in all 3 species. The phase angle of entrainment and activity duration were generally similar comparing the 2 daily activity bouts in each species. The stability of this split entrainment state was assessed by deletions of photophases on individual days, by exposure to skeleton photoperiods, and by transfer to constant darkness. As in Syrian hamsters, the one-time substitution of darkness for one 7-h photophase did not grossly alter activity patterns of Siberian hamsters but acutely disrupted the split rhythms of mice. Skeleton light pulses of progressively shorter duration did not significantly alter split entrainment patterns of either Syrian or Siberian hamsters. Both species continued to exhibit stable entrainment with activity expressed in alternate scotophases of an LD 1:5 cycle presented 4 times daily. In contrast, the split activity rhythms of mice were not maintained under skeleton pulses. In constant darkness, rhythms of Siberian hamsters remained distinctly split for a minimum of 2 cycles. Split entrainment to these novel LDLD and 4-pulse skeleton lighting regimes demonstrates a marked degree of plasticity common to the circadian systems of several rodent species and identifies novel entrainment patterns that may be reliably elicited with simple environmental manipulations. Inter- and intraspecific differences in the stability of split activity rhythms likely reflect differences in coupling interactions between the component circadian oscillators, which, adopting separate phase relations to these novel LD cycles, yield a split entrainment pattern.     

Plasticity of hamster circadian entrainment patterns depends on light intensity

The multiple oscillatory basis of the mammalian circadian pacemaker is adduced by, among other phenomena, the occurrence of split locomotor activity rhythms in rodents after prolonged exposure to constant light. More recently, split rhythms entrained to a 24h light:dark:light:dark cycle have been documented following scheduled access of hamsters to a novel running wheel or by photoperiod manipulations alone. Because the incidence of constant light-induced splitting depends on light intensity, the role of this variable was assessed in this new splitting paradigm. Male Syrian hamsters, entrained to a 14h light:10h dark cycle, were transferred to individual running wheel cages 7h after light onset. Transfer coincided with the beginning of the scotophase of a new photocycle alternating between 5h of relative dark and 7h of light. For four weeks bright photophases (approximately 350 lux) were alternated with either dim (< 0.1 lux) or completely dark (0 lux) scotophases. An additional group received moderate intensity photophases (approximately 45 lux) paired with dim scotophase illumination. For an additional four weeks, all hamsters were exposed to the same bright:dim light:dark cycle. Dim light in the scotophase significantly increased the incidence of split activity rhythms relative to that observed with completely dark scotophases. Overall wheel-running rates and activity induced by a cage change were also increased in dim light-exposed animals. Group differences largely disappeared four weeks later when hamsters previously maintained in completely dark scotophases were exposed to dim scotophases. Photophase light intensity did not affect the overall incidence of splitting, but influenced the timing of activity in the afternoon scotophase. The effects of dim illumination may be mediated in part via enhanced locomotor responses to transfer to a new cage or by changes in coupling interactions between component oscillators.     

Plasticity of Hamster Circadian Entrainment Patterns Depends on Light Intensity

The multiple oscillatory basis of the mammalian circadian pacemaker is adduced by, among other phenomena, the occurrence of split locomotor activity rhythms in rodents after prolonged exposure to constant light. More recently, split rhythms entrained to a 24h light:dark:light:dark cycle have been documented following scheduled access of hamsters to a novel running wheel or by photoperiod manipulations alone. Because the incidence of constant light-induced splitting depends on light intensity, the role of this variable was assessed in this new splitting paradigm. Male Syrian hamsters, entrained to a 14h light:10h dark cycle, were transferred to individual running wheel cages 7h after light onset. Transfer coincided with the beginning of the scotophase of a new photocycle alternating between 5h of relative dark and 7h of light. For four weeks bright photophases (～350 lux) were alternated with either dim (<0.1 lux) or completely dark (0 lux) scotophases. An additional group received moderate intensity photophases (～45 lux) paired with dim scotophase illumination. For an additional four weeks, all hamsters were exposed to the same bright:dim light:dark cycle. Dim light in the scotophase significantly increased the incidence of split activity rhythms relative to that observed with completely dark scotophases. Overall wheel-running rates and activity induced by a cage change were also increased in dim light-exposed animals. Group differences largely disappeared four weeks later when hamsters previously maintained in completely dark scotophases were exposed to dim scotophases. Photophase light intensity did not affect the overall incidence of splitting, but influenced the timing of activity in the afternoon scotophase. The effects of dim illumination may be mediated in part via enhanced locomotor responses to transfer to a new cage or by changes in coupling interactions between component oscillators.     

Daily novel wheel running reorganizes and splits hamster circadian activity rhythms.

The phenomenon of splitting of locomotor activity rhythms in constant light has implied that the mammalian circadian pacemaker is composed of multiple interacting circadian oscillators. Exposure of male Syrian hamsters to novel running wheels also induces splitting in some reports, although novel wheel running (NWR) is better known for its effects on altering circadian phase and the length of the free-running period. In three experiments, the authors confirm and extend earlier reports of split rhythms induced by NWR. Male Syrian hamsters, entrained to LD 14:10, were transferred for 6 to 11 consecutive days to darkened novel Wahmann wheels at ZT 4 and were returned to their home cages at ZT 9. All hamsters ran robustly in the novel wheels. NWR caused a marked reorganization of home cage wheel-running behavior: Activity onsets delayed progressively with each additional day of NWR. After 11 days, activity onset in the nighttime scotophase was delayed by 7 h and disappeared completely in 2 hamsters (Experiment 1). After 6 to 7 days of NWR (Experiment 2), activity onset delayed by 5 h. Transfer of hamsters to constant darkness (DD) after 7 days of NWR revealed clearly split activity rhythms: The delayed nighttime activity bout was clearly identifiable and characterized by a short duration. A second bout associated with the former time of NWR was equally distinct and exhibited a similarly short duration. These components rejoined after 3 to 5 days in DD accomplished via delays and advances of the nighttime and afternoon components, respectively. The final experiment established that rejoining of activity components could be prevented by perpetuating the light-dark:light-dark cycle used to induce split rhythms. The data suggest that NWR causes selective phase shifting of some circadian oscillators and that component oscillators interact strongly in constant darkness.     

Age-related changes in circadian responses to dark pulses

Aging involves many alterations in circadian rhythms, including a loss of sensitivity to both photic and nonphotic time signals. This study investigated the sensitivity of young and old hamsters to the phase advancing effect of a 6-h dark pulse on the locomotor activity rhythm. Each hamster was tested four times during a period of approximately 9 mo; periods of exposure to a 14-h photoperiod were alternated with the periods of exposure to constant light (20-80 lx), during which the dark pulses were administered. There was no significant difference in the phase shifts exhibited by the young (4-10 mo) and old hamsters (19-25 mo) or in the amount of wheel running activity displayed during each dark pulse. However, young hamsters had a significantly greater propensity to exhibit split rhythms immediately after the dark pulses. These results suggest that, although aging does not reduce the sensitivity of the circadian pacemaker to this nonphotic signal, it alters one property of the pacemaker, i.e., the flexibility of the coupling of its component oscillators.     

Dim nocturnal illumination alters coupling of circadian pacemakers in Siberian hamsters,<Emphasis Type="Italic"> Phodopus sungorus</Emphasis>

The circadian pacemaker of mammals comprises multiple oscillators that may adopt different phase relationships to determine properties of the coupled system. The effect of nocturnal illumination comparable to dim moonlight was assessed in male Siberian hamsters exposed to two re-entrainment paradigms believed to require changes in the phase relationship of underlying component oscillators. In experiment 1, hamsters were exposed to a 24-h light-dark-light-dark cycle previously shown to split circadian rhythms into two components such that activity is divided between the two daily dark periods. Hamsters exposed to dim illumination (<0.020 lx) during each scotophase were more likely to exhibit split rhythms compared to hamsters exposed to completely dark scotophases. In experiment 2, hamsters were transferred to winter photoperiods (10 h light, 14 h dark) from two different longer daylengths (14 h or 18 h light daily) in the presence or absence of dim nighttime lighting. Dim nocturnal illumination markedly accelerated adoption of the winter phenotype as reflected in the expansion of activity duration, gonadal regression and weight loss. The two experiments demonstrate substantial efficacy of light intensities generally viewed as below the threshold of circadian systems. Light may act on oscillator coupling through rod-dependent mechanisms.Abbreviations activity duration - DD constant dark or dim - E evening oscillator - ETV estimated testis volume - LDLD light-dark-light-dark cycle - LED light emitting diode - M morning oscillator - SCN suprachiasmatic nuclei - free-running period     

Circadian phase response curves for dark pulses in the hamster

Summary Hamsters maintained under constant illumination were exposed to 2- or 6-h pulses of darkness at various phases of their circadian activity rhythms. When presented around the time of activity onset, the pulses resulted in phase advances, and when presented toward the end of daily activity, they resulted in phase delays. Since others have shown that light pulses presented at the same phases in constant darkness cause phase shifts in the opposite directions, these results indicate that phase response curves for light and dark pulses are mirror images.Dark pulses also caused phase-dependent changes, both transient and long-lasting, in the period of the free-running rhythms, and a few pulses were immediately followed by splitting of the activity rhythms into two components. Such effects may reflect a differential responsiveness of two coupled oscillators to dark pulses.Abbreviations CT circadian time - DD constant dark - LD lightdark - LL constant light - PRC phase response curve - SD subjective day - SN subjective night - period of a circadian rhythm Supported by grants from the NSERC of Canada to B. Rusak and to G.V. Goddard. We are grateful to Dr. Goddard for his support and encouragement     

Circadian rhythms of photorefractory siberian hamsters remain responsive to melatonin

Short day lengths increase the duration of nocturnal melatonin (Mel) secretion, which induces the winter phenotype in Siberian hamsters. After several months of continued exposure to short days, hamsters spontaneously revert to the spring-summer phenotype. This transition has been attributed to the development of refractoriness of Mel-binding tissues, including the suprachiasmatic nucleus (SCN), to long-duration Mel signals. The SCN of Siberian hamsters is required for the seasonal response to winter-like Mel signals, and becomes refractory to previously effective long-duration Mel signals restricted to this area. Acute Mel treatment phase shifts circadian locomotor rhythms of photosensitive Siberian hamsters, presumably by affecting circadian oscillators in the SCN. We tested whether seasonal refractoriness of the SCN to long-duration Mel signals also renders the circadian system of Siberian hamsters unresponsive to Mel. Males manifesting free-running circadian rhythms in constant dim red light were injected with Mel or vehicle for 5 days on a 23.5-h T-cycle beginning at circadian time 10. Mel injections caused significantly larger phase advances in activity onset than did the saline vehicle, but the magnitude of phase shifts to Mel did not differ between photorefractory and photosensitive hamsters. Similarly, when entrained to a 16-h light/8-h dark photocycle, photorefractory and photosensitive hamsters did not differ in their response to Mel injected 4 h before the onset of the dark phase. Activity onset in Mel-injected hamsters was masked by light but was revealed to be significantly earlier than in vehicle-injected hamsters upon transfer to constant dim red light. The acute effects of melatonin on circadian behavioral rhythms are preserved in photorefractory hamsters.     

Entrainment of split circadian activity rhythms in hamsters

Hamsters that showed splitting of their circadian rhythms of wheel-running activity following long-term exposure to constant illumination (LL) were exposed to light-dark (LD) cycles with 2-hr dark segments, and with periods of 24.00, 24.23 or 24.72 hr. For comparison, hamsters showing nonsplit rhythms were also studied. In all cases of split rhythms, at least one of the two split components entrained to the LD cycles. In some animals, the second component continued to free-run until it merged with the entrained component, while in others, the second component also entrained to the LD cycle but maintained a stable phase angle of 6-14.5 hr relative to dark onset. These results were obtained in cases where the period of the LD cycle was shorter than that of the split rhythms and in cases where it was longer, implying that split components can be phase-advanced as well as phase-delayed by 2 hr of darkness. Three hamsters that showed stable entrainment of split rhythms were allowed to free-run in LL. The LD cycles were then reinstated, but instead of overlapping with the first component, as it did before, the dark segment was timed to overlap with the second. The entrainment patterns that ensued were similar to the ones obtained during the first LD exposure, indicating that the two split components respond to darkness in a qualitatively similar fashion. These results are further evidence that the pacemaker system underlying split circadian activity rhythms in hamsters is composed of two mutually coupled populations of oscillators that have similar properties, including a bidirectional phase response curve. Such a dual-oscillator organization may also underlie normal, or nonsplit, activity rhythms, as suggested by Pittendrigh and Daan (1976c), but the data are also compatible with the alternative view that the circadian pacemaker consists of a large number of coupled oscillators, which only dissociate into two separate populations in some animals under conditions of moderate LL intensity.     

Hamsters Running on Time: Is the Lateral Habenula a Part of the Clock?

Previous anatomical and physiological studies have implicated the lateral habenula, and especially its medial division (LHbM), as a candidate component of the circadian timing system in rodents. We assayed lateral habenula rhythmicity in rodents using c‐FOS immunohistochemistry and found a robust rhythm in immunoreactive cell counts in the LHbM, with higher counts during the dark phase of a light‐dark (LD) cycle and during subjective night in constant darkness. We have also observed an obvious asymmetry of c‐FOS expression in the LHbM of behaviorally “split” hamsters in constant light, but only during their active phase (when they were running in wheels). Locomotor activity rhythms appear to be regulated by the suprachiasmatic nucleus (SCN) via multiple output pathways, one of which might be diffusible while the other might be neural, involving the lateral habenula.     

Hamsters running on time: is the lateral habenula a part of the clock?

Previous anatomical and physiological studies have implicated the lateral habenula, and especially its medial division (LHbM), as a candidate component of the circadian timing system in rodents. We assayed lateral habenula rhythmicity in rodents using c-FOS immunohistochemistry and found a robust rhythm in immunoreactive cell counts in the LHbM, with higher counts during the dark phase of a light-dark (LD) cycle and during subjective night in constant darkness. We have also observed an obvious asymmetry of c-FOS expression in the LHbM of behaviorally "split" hamsters in constant light, but only during their active phase (when they were running in wheels). Locomotor activity rhythms appear to be regulated by the suprachiasmatic nucleus (SCN) via multiple output pathways, one of which might be diffusible while the other might be neural, involving the lateral habenula.     

Running activity mediates the phase-advancing effects of dark pulses on hamster circadian rhythms

Summary Pulses of darkness can phase-shift the circadian activity rhythms of hamsters,Mesocricetus auratus, kept in constant light. Dark pulses under these conditions alter photic input to the circadian system, but they also commonly trigger wheel-running activity. This paper investigates the contribution of running activity to the phase-shifting effects of dark pulses. A first experiment showed that running activity by itself can phaseshift rhythms in constant light. Hamsters were induced to run by being confined to a novel wheel for 3–5 h. When this was done at circadian times (CT) 0, 6, and 9, the mean steady-state phase-shifts were 0.6 h, 3.5 h, and 2.3 h, respectively. The latter two values are at least as large as those previously obtained with dark pulses of similar durations and circadian phases. A second experiment showed that restricting the activity of hamsters during 3-h dark pulses at CT 9 reduces the amplitude of the phase-shifts. Unrestrained animals phase-advanced by 1.1 h, but this shift was halved in animals whose wheel was locked, and completely abolished in animals confined to nest boxes during the dark pulse. Activity restriction in itself (without dark pulses) had only minimal phase-delaying effects on free-running rhythms when given between ca. CT 10 and CT 13. These results support the idea that, in hamsters at least, dark pulses affect the circadian system mostly by altering behavioural states rather than by altering photic input to the internal clock.Abbreviations CT circadian time - DD constant darkness - LD light-dark - LL constant light - PRC phase response curve - period of rhythm     

Non-parametric photic entrainment of Djungarian hamsters with different rhythmic phenotypes

To investigate the role of non-parametric light effects in entrainment, Djungarian hamsters of two different circadian phenotypes were exposed to skeleton photoperiods, or to light pulses at different circadian times, to compile phase response curves (PRCs). Wild-type (WT) hamsters show daily rhythms of locomotor activity in accord with the ambient light/dark conditions, with activity onset and offset strongly coupled to light-off and light-on, respectively. Hamsters of the delayed activity onset (DAO) phenotype, in contrast, progressively delay their activity onset, whereas activity offset remains coupled to light-on. The present study was performed to better understand the underlying mechanisms of this phenomenon. Hamsters of DAO and WT phenotypes were kept first under standard housing conditions with a 14:10 h light–dark cycle, and then exposed to skeleton photoperiods (one or two 15-min light pulses of 100 lx at the times of the former light–dark and/or dark–light transitions). In a second experiment, hamsters of both phenotypes were transferred to constant darkness and allowed to free-run until the lengths of the active (α) and resting (ρ) periods were equal (α:ρ = 1). At this point, animals were then exposed to light pulses (100 lx, 15 min) at different circadian times (CTs). Phase and period changes were estimated separately for activity onset and offset. When exposed to skeleton-photoperiods with one or two light pulses, the daily activity patterns of DAO and WT hamsters were similar to those obtained under conditions of a complete 14:10 h light–dark cycle. However, in the case of giving only one light pulse at the time of the former light–dark transition, animals temporarily free-ran until activity offset coincided with the light pulse. These results show that photic entrainment of the circadian activity rhythm is attained primarily via non-parametric mechanisms, with the “morning” light pulse being the essential cue. In the second experiment, typical photic PRCs were obtained with phase delays in the first half of the subjective night, phase advances in the second half, and a dead zone during the subjective day. ANOVA indicated no significant differences between WT and DAO animals despite a significantly longer free-running period (tau) in DAO hamsters. Considering the phase shifts induced around CT0 and the different period lengths, it was possible to model the entrainment patterns of both phenotypes. It was shown that light-induced phase shifts of activity offset were sufficient to compensate for the long tau in WT and DAO hamsters, thus enabling a stable entrainment of their activity offsets to be achieved. With respect to activity onsets, phase shifts were sufficient only in WT animals; in DAO hamsters, activity onset showed increasing delays. The results of the present paper clearly demonstrate that, under laboratory conditions, the non-parametric component of light and dark leads to circadian entrainment in Djungarian hamsters. However, a stable entrainment of activity onset can be achieved only if the free-running period does not exceed a certain value. With longer tau values, hamsters reveal a DAO phenotype. Under field conditions, therefore, non-photic cues/zeitgebers must obviously be involved to enable a proper circadian entrainment.     

Lesions of the thalamic intergeniculate leaflet alter hamster circadian rhythms

We have investigated the effects of destruction of the geniculo-hypothalamic tract (GHT) on the circadian system of golden hamsters. In the first experiment, intact hamsters were housed in constant darkness, and phase shifts in running-wheel activity rhythms were assessed following 15-min light pulses administered at circadian time (CT) 12 (defined as the beginning of activity), CT 14, CT 18, and CT 20. Responses to light pulses at the same CTs were then reassessed after GHT lesions. Hamsters with complete lesions showed decreases in phase advances caused by light pulses at CT 18 and CT 20. Phase delays elicited by light at CT 12 and CT 14 were not altered. In a second study, intact and GHT-ablated hamsters housed in constant light received 6-hr dark pulses at various CTs. Hamsters with complete GHT ablation showed smaller advances than controls to dark pulses centered on CT 8-10. After 110 days in constant light, 7 of 10 intact hamsters showed splitting of their activity rhythms into two components, while only 1 of the 8 similarly treated ablated hamsters displayed dissociated activity components. Ablated hamsters had significantly shorter free-running periods during the first 35 days of exposure to constant light than did the intact hamsters. These results demonstrate that destruction of the GHT in the hamster alters phase shifting in response to periods of light or dark, and they indicate a role for the GHT in mediating several photic effects on the circadian system.     

Circadian Rhythms of Enzymes' Activity in Mice Tissues During Exposure to Continuous Illumination

Activity rhythms of enzymes were determined in various tissues of C57BL/6J male mice. The determinations were carried out on mice which were kept in 14 hr light: 10 hr dark regimen, and on day 2, day 5 and day 21 during exposure to continuous illumination. Locomotor activity rhythms were followed in light: dark and up to the seventh day in constant light. All the activities exhibited a significant circadian rhythm in the light: dark regimen. During the exposure to continuous illumination, the locomotor activity exhibit a free running circadian rhythm with a consistent 24 hr and 40 min, major period component. At the same time recording the rhythms of enzyme activity; enzymes exhibited various formats of response which differed from those of the locomotor activity. The results suggest that rhythms of enzyme activity, as well as the desynchronization of the rhythms, are not enzyme specific.     

Circadian activity pattern in the stable fly, Stomoxys calcitrans

Abstract. Activity rhythms of Stomoxys calcitrans (L.) were assessed under constant conditions using actographs.In LD 12:12 h, activity was almost completely diurnal, had a 24 h period, and peaked 4 h after lights-on.In constant darkness, activity peaked in the early afternoon and the period of the rhythm was c. 26 h, indicating a circadian pacemaker.A delay in feeding shortened the free-running period in constant dark to c. 24 h.In constant light, flies were arrhythmic.Starvation also affected activity, with 24- and 48-h starved flies being as much as 20 times more active than recently fed flies.     

Phase angle difference alters coupling relations of functionally distinct circadian oscillators revealed by rhythm splitting

The interactions (i.e., coupling) between multiple oscillators of a circadian system determine basic properties of the integrated pacemaker. Unfortunately, there are few experimental models to investigate the putative interactions of functionally defined oscillators comprising the mammalian circadian pacemaker. Here the authors induce in hamsters a novel circadian entrainment pattern that is characterized by the daily expression of robust wheel-running activity in each scotophase of a 24-h light:dark:light:dark cycle. The daily activity bouts are mediated by 2 circadian oscillators, here designated "daytime" and "nighttime," that have been temporally dissociated under this light regime. To assess the phase dependence of interactions between oscillatory components, the phase relationship of the 2 daily scotophases was manipulated over a 4-h range, and the timing of activity of the daytime and nighttime components under entrained and probe conditions was examined. The average phase angle of entrainment and the day-to-day variability of activity onset of each activity component depended on the phase relationship of the respective scotophases and not on whether the component occurred in the daytime or the nighttime. Short-term denial of wheel access subsequently influenced amount and duration of wheel running but not timing of its onset, suggesting that only the former measures depend on a homeostatic mechanism sensitive to the time elapsed since prior intense running. Replacement of individual photophases with darkness revealed phase attraction between oscillators that was not dependent on the phase relationship of component oscillators but differed for daytime versus nighttime activity components. Entrainment patterns shown here cannot be accounted for by only nonparametric actions of light. Instead, the phase-dependent interactions of oscillators strongly influence entrainment properties, whereas intrinsic functional differences in dissociated oscillators apparently influence their attraction in darkness. This model system may be ideal for identifying genomic and physiological factors that mediate these interactions and thus contribute importantly to system properties of the mammalian circadian clock.     

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.