GABA(B) receptor activation in the suprachiasmatic nucleus of diurnal and nocturnal rodents

Diurnal (day-active) and nocturnal (night-active) animals have very different daily activity patterns. We recently demonstrated that the suprachiasmatic nucleus (SCN) responds to GABAergic stimulation differently in diurnal and nocturnal animals. Specifically, GABAA receptor activation with muscimol during the subjective day causes phase delays in diurnal grass rats while producing phase advances in nocturnal hamsters. The aim of the following experiments was to determine if diurnal and nocturnal animals differ in their response to GABAB receptor activation in the SCN. Baclofen, a GABAB receptor agonist, was microinjected into the SCN region of grass rats or hamsters under free-running conditions and phase alterations were analyzed. Changes in phase were not detected after baclofen treatment during the subjective day in either grass rats or hamsters. During the night, however, GABAB receptor activation significantly decreased the ability of light to induce phase delays in grass rats. Taken together with previous data from our laboratory, these results demonstrate that, in both hamsters and grass rats, GABAB receptor activation in the SCN significantly affects circadian phase during the night, but not during the day.     

Light and GABA)(A) receptor activation alter period mRNA levels in the SCN of diurnal Nile grass rats

We examined Period (Per) mRNA rhythms in the suprachiasmatic nucleus (SCN) of a diurnal rodent and assessed how phase-shifting stimuli acutely affect SCN Per mRNA using semiquantitative in situ hybridization. First, Per1 and Per2 varied rhythmically in the SCN over the course of one circadian cycle in constant darkness: Per1 mRNA was highest in the early to mid-subjective day, while Per2 mRNA levels peaked in the late subjective day. Second, acute light exposure in the early subjective night significantly increased both Per1 and Per2 mRNA. Third, Per2 but not Per1 levels decreased 1 and 2 h after injection of the gamma-aminobutyric acid (GABA)(A) receptor agonist muscimol into the SCN during the subjective day. Fourth, muscimol also reduced the light-induced Per2 in the early subjective night, but Per1 induction by light was not significantly affected. Consistent with previous studies, these data demonstrate that diurnal and nocturnal animals show very similar daily patterns of Per mRNA and light-induced Per increases in the SCN. As with light, muscimol alters circadian phase, and daytime phase alterations induced by muscimol are associated with significant decreases in Per2 mRNA. In diurnal animals, muscimol-induced decreases in Per are associated with phase delays rather than advances. The direction of the daytime phase shift may be determined by the relative suppression of Per1 vs. Per2 in SCN cells. As in nocturnal animals, changes in Per1 and Per2 mRNA by photic and non-photic stimuli appear to be associated with circadian phase alteration.     

Fos rhythms in the hypothalamus of Rattus and Arvicanthis that exhibit nocturnal and diurnal patterns of rhythmicity

This study compared patterns of Fos expression within the suprachiasmatic nucleus (SCN), the region immediately dorsal to the SCN (the lower subparaventricular zone, LSPV), and the supraoptic nucleus (SON) of grass rats (Arvicanthis niloticus) and lab rats (Rattus norvegicus). Among grass rats we also compared individuals exhibiting nocturnal and diurnal patterns of wheel running. In the SCN of both groups of grass rats, as well as laboratory rats, Fos was elevated during the light compared to the dark portions of the day, and was expressed in 7–12% of cells containing vasoactive intestinal polypeptide (VIP). Fos was higher in the LSPV during the night compared to the day in both forms of grass rats but not in laboratory rats. In the SON, Fos rose from day to night in the diurnal grass rats and in laboratory rats, but not in nocturnal grass rats. These patterns are consistent with the hypothesis that VIP cells in the SCN function similarly in nocturnal and diurnal rodents, but that the SON and the region dorsal to the SCN are associated with intra and interspecific differences in rhythmicity, respectively.     

Suprachiasmatic nucleus projections to the paraventricular thalamic nucleus in nocturnal rats (Rattus norvegicus) and diurnal nile grass rats (Arviacanthis niloticus)

The circadian pacemaker of the suprachiasmatic nucleus (SCN) is likely to control the timing of the sleep–wake cycle in mammals by modulating the daily activity patterns of brain regions important in sleep and wakefulness. One such brain region is the paraventricular nucleus of the thalamus (PVT). In both nocturnal rats and the diurnal rodent Arvicanthis niltoicus (Nile grass rat), expression of Fos (the product of the immediate-early gene c-fos) in the PVT increases at times of day when the animals are most active. To compare the projections of the SCN to the PVT in these two species, the retrograde tracer cholera toxin (β subunit; CTβ) was microinjected into the PVT and the SCN was examined to identify labeled neurons. Further, the PVT-projecting SCN cells containing either arginine vasopressin (AVP) or gastrin releasing peptide (GRP) were also compared between species. In both nocturnal rats and diurnal Nile grass rats, the SCN sends a substantial projection to the PVT. In both species, many PVT-projecting SCN neurons contain AVP, and few contain GRP. Other work has shown that some AVP-containing neurons of the SCN function differently in rats and Nile grass rats. Projections from functionally distinct SCN neurons to the PVT may contribute to the difference in the temporal distribution of sleep and wakefulness seen between these two species.     

Shedding light on circadian clock resetting by dark exposure: differential effects between diurnal and nocturnal rodents

The master circadian clock in mammals, located in the suprachiasmatic nuclei (SCN) of the hypothalamus, is entrained by light and behavioural stimulation. In addition, the SCN can be reset by dark pulses in nocturnal rodents under constant light conditions. Here, the shifting effects of a dark pulse on the SCN clock were detailed at both a behavioural and molecular level in a nocturnal rodent (Syrian hamster), and were compared to those of a diurnal rodent (Arvicanthis ansorgei). Four-hour dark pulses led to phase advances in the circadian rhythm of locomotor activity from subjective midday to dusk in hamsters, but from subjective dusk to midnight in Arvicanthis. Moreover, dark pulses had no resetting effect during the middle of the subjective night in hamsters, while such a dead shifting zone occurred during most of the subjective day in Arvicanthis. The behavioural phase advances in both hamsters and Arvicanthis were most often accompanied by marked downregulation of the clock genes Per1 and/or Per2 in the SCN, and also by changes in the transforming growth factor-alpha expression, a neuropeptide that suppresses daytime activity in nocturnal mammals. Despite that both hamsters and Arvicanthis showed dark-induced phase advances at circadian time-12, Per1 gene and its protein PER1 were downregulated in Arvicanthis but not in hamsters. Altogether these results show that dark resetting of the SCN is always associated with downregulation of Per1 and/or Per2 expression, and mostly occurs during resting. Thus, the circadian window of sensitivity to dark differs between nocturnal and diurnal rodents.     

Serotonergic activation potentiates light resetting of the main circadian clock and alters clock gene expression in a diurnal rodent

The main circadian clock, localized in the suprachiasmatic nuclei (SCN) in mammals, can be synchronized by light and non-photic factors such as serotonergic cues. In nocturnal rodents, injections during the subjective day of the 5-HT1A/7 receptor agonist 8-hydroxy-2-(di-n-propylamino) tetralin (8-OH-DPAT) or its positive enantiomer, induce behavioral phase-advances in correlation with decreased expression of two clock genes, Per1/2. In addition, 8-OH-DPAT and the selective serotonin reuptake inhibitor fluoxetine reduce light-induced phase-shifts during the subjective night. Beside the chronobiotic effects of serotonin, changes of serotonergic activity in humans have been involved in mood disorders, that are often associated with alterations in circadian rhythmicity. To get insights into the circadian role of serotonin in diurnal species, we investigated its modulation of the SCN in Arvicanthis ansorgei housed in constant darkness. In striking contrast to nocturnal rodents, daily serotonin content in Arvicanthis SCN peaked during daytime while the sensitivity window of its SCN to (+)8-OH-DPAT occurred essentially during the subjective night. Moreover, fluoxetine produced behavioral phase-advances at circadian time (CT) 0 and CT12. Expression of Per1/2, Rev-erbalpha/beta and Roralpha/beta in the SCN was not modified after fluoxetine or (+)8-OH-DPAT injection. Furthermore, both treatments enhanced light-induced phase-advances and delays. Light responses of Per1 and Rorbeta expression at CT0 and those of Per2 and Rev-erbalpha at CT12 were markedly altered by serotonergic activation. The present findings demonstrate that the serotonergic modulation of the SCN clock appears to differ between nocturnal species and the diurnal Arvicanthis. The potentiating effects of fluoxetine on light resetting in a diurnal rodent may be clinically relevant.     

Photic regulation of c-fos expression in neural components governing the entrainment of circadian rhythms

The rapid and transient induction of the proto-oncogene c-fos in mature neurons within the brain occurs in response to a variety of extracellular stimuli. To determine whether lighting conditions influence c-fos gene expression in the primary neural structures mediating the photoentrainment and generation of mammalian circadian rhythms, the expression of the c-fos protein (Fos) and related proteins in the retina and suprachiasmatic nuclei (SCN) of the anterior hypothalmus was examined immunohistochemically in rats exposed to a light-dark cycle of 12 h of light and 12 h of darkness (LD 12:12), constant light (LL), or constant dark (DD). The retina exhibited clear light-dark differences in the expression of Fos protein(s), such that immunopositive nuclei were readily evident during exposure to light (i.e., during the day of diurnal lighting or in LL), but were absent during exposure to darkness. In the SCN, the distribution of Fos immunoreactivity within specific subfields was differentially affected by photic conditions. Following exposure to light, a dense population of Fos-immunopositive cells was found in close association with the immunohistochemically distinct cell and fiber populations distinguishing the ventrolateral subfield of the SCN. In dark-exposed animals, Fos-immunoreactive profiles were distributed throughout the SCN in areas coextensive with the immunohistochemical localization of peptidergic neural elements in both the ventrolateral and dorsomedial subfields. As a consequence of this light-dark difference in the distribution of Fos immunoreactivity, the density of labeled cells was increased within the ventrolateral SCN, but was decreased within the dorsomedial subfield, as a result of exposure to light versus darkness. In the absence of photic time cues, temporal variation in the pattern of Fos immunostaining in the SCN, or within specific subfields of the nucleus, was evident only within dorsomedial SCN during exposure to LL, such that the density of immunopositive cells was greater during the subjective day than during the subjective night. These data demonstrate that light stimulation causes an increase in the expression of Fos protein(s) in the retina and within the ventrolateral, but not the dorsomedial, subfield of the SCN. The inductive effect of light on Fos expression within the retina and the ventrolateral or retinorecipient subfield of the SCN suggests that Fos protein(s) may play a role in the transduction of light signals by the primary neural components governing the generation and photoentrainment of circadian rhythms in mammals.     

Electrophysiology of optic nerve input to suprachiasmatic nucleus neurons in rats and degus

Neurons in the mammalian suprachiasmatic nucleus (SCN), the principal pacemaker of the circadian system, receive direct retinal input. Some SCN neurons respond to retinal illumination or optic nerve stimulation with changes in firing rates. In nocturnal rodents, retinal illumination increases firing rates of a large majority and decreases firing rates of a minority of responsive neurons. In two species of diurnal rodent, these proportions are altered or even reversed. Since retinal input to the SCN has been reported to involve release of the excitatory neurotransmitter glutamate, the mechanism mediating suppressions is unknown. We studied responses of neurons in SCN slices from diurnal degus and nocturnal rats to optic nerve stimulation. To test whether suppressions are mediated indirectly by release of the inhibitory neurotransmitter GABA from SCN neurons that are first activated by glutamate release, we attempted to block suppressions by adding to the bath either APV, an antagonist for excitatory glutamate receptors, or bicuculline, a GABA_A receptor antagonist. If glutamate is the only neurotransmitter released by optic nerves in the SCN, APV should prevent both activations and suppressions in response to optic nerve stimulation. We found that APV had little effect on suppressions although it effectively blocked activations. Bicuculline blocked most suppressions. These findings are inconsistent with a model in which the retina provides only excitatory glutamate input to the SCN via NMDA receptors. Since some retinal fibers in adult mammals contain GABA, it is possible that the retinal innervation of the SCN includes both glutamate- and GABA-containing axons.     

Development of the light sensitivity of the clock genes Period1 and Period2, and immediate-early gene c-fos within the rat suprachiasmatic nucleus

The molecular mechanism underlying circadian rhythmicity within the suprachiasmatic nuclei (SCN) of the hypothalamus has two light-sensitive components, namely the clock genes Per1 and Per2 . Besides, light induces the immediate-early gene c-fos . In adult rats, expression of all three genes is induced by light administered during the subjective night but not subjective day. The aim of the present study was to ascertain when and where within the SCN the photic sensitivity of Per1 , Per2 and c-fos develops during early postnatal ontogenesis. The specific aim was to find out when the circadian clock starts to gate photic sensitivity. The effect of a light pulse administered during either the subjective day or the first or second part of the subjective night on gene expression within the rat SCN was determined at postnatal days (P) 1, 3, 5 and 10. Per1 , Per2 and c-fos mRNA levels were assessed 30 min, 1 and 2 h after the start of each light pulse by in situ hybridization histochemistry. Expression of Per1 and c-fos was light responsive from P1, and the responses began to be gated by the circadian clock at P3 and P10, respectively. Expression of Per2 was only slightly light responsive at P3, and the response was not fully gated until P5. These data demonstrate that the light sensitivity of the circadian clock develops gradually during postnatal ontogenesis before the circadian clock starts to control the response. The photoinduction of the clock gene Per2 develops later than that of Per1 .     

Suprachiasmatic nucleus projections to the paraventricular thalamic nucleus in nocturnal rats (Rattus norvegicus) and diurnal nile grass rats (Arviacanthis niloticus)

The circadian pacemaker of the suprachiasmatic nucleus (SCN) is likely to control the timing of the sleep-wake cycle in mammals by modulating the daily activity patterns of brain regions important in sleep and wakefulness. One such brain region is the paraventricular nucleus of the thalamus (PVT). In both nocturnal rats and the diurnal rodent Arvicanthis niltoicus (Nile grass rat), expression of Fos (the product of the immediate-early gene c-fos) in the PVT increases at times of day when the animals are most active. To compare the projections of the SCN to the PVT in these two species, the retrograde tracer cholera toxin (beta subunit; CTbeta) was microinjected into the PVT and the SCN was examined to identify labeled neurons. Further, the PVT-projecting SCN cells containing either arginine vasopressin (AVP) or gastrin releasing peptide (GRP) were also compared between species. In both nocturnal rats and diurnal Nile grass rats, the SCN sends a substantial projection to the PVT. In both species, many PVT-projecting SCN neurons contain AVP, and few contain GRP. Other work has shown that some AVP-containing neurons of the SCN function differently in rats and Nile grass rats. Projections from functionally distinct SCN neurons to the PVT may contribute to the difference in the temporal distribution of sleep and wakefulness seen between these two species.     

Fos rhythms in the hypothalamus of Rattus and Arvicanthis that exhibit nocturnal and diurnal patterns of rhythmicity

This study compared patterns of Fos expression within the suprachiasmatic nucleus (SCN), the region immediately dorsal to the SCN (the lower subparaventricular zone, LSPV), and the supraoptic nucleus (SON) of grass rats (Arvicanthis niloticus) and lab rats (Rattus norvegicus). Among grass rats we also compared individuals exhibiting nocturnal and diurnal patterns of wheel running. In the SCN of both groups of grass rats, as well as laboratory rats, Fos was elevated during the light compared to the dark portions of the day, and was expressed in 7-12% of cells containing vasoactive intestinal polypeptide (VIP). Fos was higher in the LSPV during the night compared to the day in both forms of grass rats but not in laboratory rats. In the SON, Fos rose from day to night in the diurnal grass rats and in laboratory rats, but not in nocturnal grass rats. These patterns are consistent with the hypothesis that VIP cells in the SCN function similarly in nocturnal and diurnal rodents, but that the SON and the region dorsal to the SCN are associated with intra and interspecific differences in rhythmicity, respectively.     

Gastrin-releasing peptide phase-shifts suprachiasmatic nuclei neuronal rhythms in vitro.

The main mammalian circadian pacemaker is located in the suprachiasmatic nuclei (SCN) of the hypothalamus. Gastrin-releasing peptide (GRP) and its receptor (BB(2)) are synthesized by rodent SCN neurons, but the role of GRP in circadian rhythm processes is unknown. In this study, we examined the phase-resetting actions of GRP on the electrical activity rhythms of hamster and rat SCN neurons in vitro. In both rat and hamster SCN slices, GRP treatment during the day did not alter the time of peak SCN firing. In contrast, GRP application early in the subjective night phase-delayed, whereas similar treatment later in the subjective night phase-advanced the firing rate rhythm in rat and hamster SCN slices. These phase shifts were completely blocked by the selective BB(2) receptor antagonist, [d-Phe(6), Des-Met(14)]-bombesin 6-14 ethylamide. We also investigated the temporal changes in the expression of genes for the BB(1) and BB(2) receptors in the rat SCN using a quantitative competitive RT-PCR protocol. The expression of the genes for both receptors was easily detected, but their expression did not vary over the diurnal cycle. These data show that GRP phase-dependently phase resets the rodent SCN circadian pacemaker in vitro apparently via the BB(2) receptor. Because this pattern of phase shifting resembles that of light on rodent behavioral rhythms, these results support the contention that GRP participates in the photic entrainment of the rodent SCN circadian pacemaker.     

Photically responsive neurons in the hypothalamus of a diurnal ground squirrel

The suprachiasmatic nuclei (SCN) function as a circadian pacemaker. Entrainment to the external light-dark cycle is mediated by the retina which gives rise to both direct and indirect projections to the SCN. The hypothalamic targets of the retinohypothalamic tract (RHT) were investigated in thirteen-lined ground squirrels (Spermophilus tridecemlineatus) following WGA-HRP injections. The results indicate that retinal fibers project to the entire SCN. Extracellular single-unit activity was recorded in and near the SCN of thiopental sodium anesthetized squirrels while the eyes were photically stimulated. A small population of hypothalamic cells were responsive to retinal illumination. About half of these cells were activated by light while the others were light-suppressed. The majority of these cells responded in a sustained way to light pulses. Light intensities of at least 1000 lux appeared necessary to induce a sustained response to light. No differences in light responsiveness were observed between visual cells inside and outside the SCN. The visual properties of SCN cells have previously been investigated in hamsters and rats, both nocturnal species. Hypothalamic cells in all 3 species were similar in that they showed predominantly sustained responses to retinal illumination. The diurnal squirrel differed from the other two species in that there was a higher proportion of photically suppressed cells in the squirrel, and in that higher light intensities were required to stimulate photically responsive neurons.     

Light-induced Fos expression in the suprachiasmatic nucleus of the four-striped field mouse,Rhabdomys pumilio: A southern African diurnal rodent

Previous studies have suggested that nocturnal and diurnal species of rodents differ in their circadian responses to light including phase shifts and early gene expression. Rhabdomys pumilio, the four-striped field mouse, is diurnal both in nature and in the laboratory. We studied in this species the response of the suprachiasmatic nucleus (SCN) to light stimuli at different time periods using light-induced expression of Fos as marker of neuronal activity. Fos induction in the SCN was investigated using immunohistochemistry and quantitative image analysis. The animals were exposed to a 15 min light pulse with monochromatic green light at different circadian times throughout a 24-h cycle. Animals maintained in constant darkness served as controls. R. pumilio exhibited an endogenous Fos rhythm in the SCN during constant darkness with highest expression during the subjective day at circadian time (CT) 2 and CT10. Photic stimulation resulted in significant Fos induction in the SCN at CT6, CT14, CT18 and CT22, compared to controls kept in constant darkness, with a peak of expression at CT22, i.e. during late subjective night, mainly due to expression in the ventral SCN. In tract tracing experiments based on the use of cholera toxin subunit B, we found that retinal fibres innervate mainly the contralateral ventral SCN. The intergeniculate leaflet received bilateral retinal innervation with overlap between ipsilateral and contralateral fibres. Altogether the data show that the rodent R. pumilio is a unique diurnal model for chronobiological studies.     

Light-induced Fos expression in the suprachiasmatic nucleus of the four-striped field mouse, Rhabdomys pumilio: A southern African diurnal rodent

Previous studies have suggested that nocturnal and diurnal species of rodents differ in their circadian responses to light including phase shifts and early gene expression. Rhabdomys pumilio, the four-striped field mouse, is diurnal both in nature and in the laboratory. We studied in this species the response of the suprachiasmatic nucleus (SCN) to light stimuli at different time periods using light-induced expression of Fos as marker of neuronal activity. Fos induction in the SCN was investigated using immunohistochemistry and quantitative image analysis. The animals were exposed to a 15 min light pulse with monochromatic green light at different circadian times throughout a 24-h cycle. Animals maintained in constant darkness served as controls. R. pumilio exhibited an endogenous Fos rhythm in the SCN during constant darkness with highest expression during the subjective day at circadian time (CT) 2 and CT10. Photic stimulation resulted in significant Fos induction in the SCN at CT6, CT14, CT18 and CT22, compared to controls kept in constant darkness, with a peak of expression at CT22, i.e. during late subjective night, mainly due to expression in the ventral SCN. In tract tracing experiments based on the use of cholera toxin subunit B, we found that retinal fibres innervate mainly the contralateral ventral SCN. The intergeniculate leaflet received bilateral retinal innervation with overlap between ipsilateral and contralateral fibres. Altogether the data show that the rodent R. pumilio is a unique diurnal model for chronobiological studies.     

Circadian and photic modulation of daily rhythms in diurnal mammals

The temporal niche that an animal occupies includes a coordinated suite of behavioral and physiological processes that set diurnal and nocturnal animals apart. The daily rhythms of the two chronotypes are regulated by both the circadian system and direct responses to light, a process called masking. Here we review the literature on circadian regulations and masking responses in diurnal mammals, focusing on our work using the diurnal Nile grass rat (Arvicanthis niloticus) and comparing our findings with those derived from other diurnal and nocturnal models. There are certainly similarities between the circadian systems of diurnal and nocturnal mammals, especially in the phase and functioning of the principal circadian oscillator within the hypothalamic suprachiasmatic nucleus (SCN). However, the downstream pathways, direct or indirect from the SCN, lead to drastic differences in the phase of extra‐SCN oscillators, with most showing a complete reversal from the phase seen in nocturnal species. This reversal, however, is not universal and in some cases the phases of extra‐SCN oscillators are only a few hours apart between diurnal and nocturnal species. The behavioral masking responses in general are opposite between diurnal and nocturnal species, and are matched by differential responses to light and dark in several retinorecipient sites in their brain. The available anatomical and functional data suggest that diurnal brains are not simply a phase‐reversed version of nocturnal ones, and work with diurnal models contribute significantly to a better understanding of the circadian and photic modulation of daily rhythms in our own diurnal species.     

Effects of intergeniculate leaflet lesions on circadian rhythms in Octodon degus

The intergeniculate leaflet (IGL) modulates photic and nonphotic entrainment of circadian rhythms in nocturnal species, but nothing is known about its role in diurnal species. We investigated the significance of the IGL for circadian rhythm function in the diurnal rodent, Octodon degus, by determining the effects of bilateral electrolytic IGL lesions (IGL(X)) on: (i) photic entrainment; (ii) reentrainment rates to photic cues following a 6-h phase advance of the light-dark (LD) cycle; (iii) reentrainment rates to nonphotic social and photic cues following a 6-h phase advance of the LD cycle; and (iv) the circadian period (tau) of the activity rhythm in constant darkness (DD). IGL(X) significantly lengthened the duration (alpha) of the entrained activity rhythm and produced a significantly earlier phase of activity onset under entrained (LD 12:12) conditions, but did not change phase of activity offset, rhythm amplitude or mean daily activity levels. IGL(X) failed to modify tau of free-running activity rhythms in DD or alter reentrainment rates of circadian rhythms to nonphotic social and photic cues or photic cues alone. Thus, the IGL modulates two parameters of photic entrainment, but is not necessary for reentrainment to either nonphotic social or photic cues. Our results contribute to the growing comparative database on the neural mechanisms controlling circadian rhythms and indicate that the role of the IGL varies across species with no apparent relationship between diurnality-nocturnality and circadian function.     

Vasoactive intestinal polypeptide (VIP) phase-shifts the rat suprachiasmatic nucleus clock in vitro

In mammals, the principal circadian pacemaker is housed in the hypothalamic suprachiasmatic nuclei (SCN). The SCN exhibit high levels of vasoactive intestinal polypeptide (VIP) immunoreactivity and two of the three VIP receptors, VPAC(2) and PAC(1), are found in the rat SCN. However, the role of VIP in the SCN remains unclear. In this study, we examined the phase-resetting actions of VIP and selective VIP receptor agonists on the electrical activity rhythm of rat SCN neurons in vitro. Application of VIP during the subjective day did not shift the peak in the firing rate rhythm. However, VIP treatment during the early or late subjective night evoked a small phase delay or a large phase advance, respectively. The phase-advancing effect of VIP was reproduced by the novel VPAC(2) receptor agonist RO 25-1553, but not by pituitary adenylate cyclase-activating peptide (a potent PAC(1) receptor agonist), or by [K15,R16,L27]VIP(1-7)/GRF(8-27), a novel, selective VPAC(1) receptor agonist. These data show that VIP phase-dependently phase-resets the rodent SCN pacemaker in vitro, presumably via the VPAC(2) receptor. As the pattern of phase-shifting evoked by VIP and RO 25-1553 resembles the phase-resetting actions of light on rodent behavioural rhythms, these data support a role for VIP and the VPAC(2) receptor in photic entrainment of the rodent circadian pacemaker.     

Removal of the olfactory bulbs delays photic reentrainment of circadian activity rhythms and modifies the reproductive axis in male Octodon degus

The diurnal rodent, Octodon degus, exhibits robust sex differences in several circadian measures, including circadian period (τ) and reentrainment rates to photic and nonphotic (social) zeitgebers. The neural substrates underlying such physiological differences remain unknown. In female degus, olfactory bulbectomies (BX) inhibit socially-facilitated reentrainment, but do not alter photic reentrainment, entrained measures, or τ in constant darkness (DD). This experiment investigated the effects of BX in male degus on (i) photic reentrainment rates of circadian rhythms following a 6-h phase advance of the light–dark (LD) cycle; (ii) photic entrainment; (iii) τ of free-running activity rhythms in DD; and (iv) body weight, paired testis weight, and the reproductive hormones, testosterone, androstenedione and follicle stimulating hormone (FSH). BX significantly delayed photic reentrainment rates. They did not, however, modify τ, the phase of activity onset or offset, amplitude or duration (α) of the activity rhythm, mean daily locomotor activity levels, or body weight. FSH, testosterone and androstenedione were unaffected by BX, whereas paired testis weights were significantly greater in BX degus compared with shams. Thus, the olfactory bulbs influence photic reentrainment of circadian rhythms and modestly affect the reproductive axis in male degus. Our results suggest that the olfactory bulbs may be a neural source of observed sex differences in photic reentrainment in degus, and highlight interspecies variation in the olfactory bulbs' effects on entrained and free-running circadian rhythms and on reproduction.     

N-Methyl-D-aspartate microinjected into the suprachiasmatic nucleus mimics the phase-shifting effects of light in the diurnal Nile grass rat (Arvicanthis niloticus)

Mammals exhibit circadian rhythms in behavior generated by the suprachiasmatic nucleus (SCN). Exposure to light synchronizes the circadian clock to the environmental light:dark cycle through the release of glutamate into the SCN. In nocturnal animals such as Syrian hamsters, direct application of NMDA to the SCN results in phase shifts similar to those produced by exposure to light. This study was designed to determine if light phase shifts the circadian pacemaker of diurnal Nile grass rats (Arvicanthis niloticus) housed in constant darkness by acting on NMDA-type glutamate receptors in the suprachiasmatic nucleus (SCN). N-Methyl-D-aspartate (NMDA; 0, 1, 10, 50, and 100 mM) was administered through guide cannulae aimed at the SCN at circadian times when light induces phase shifts. Maximal phase delays were attained with 50 mM NMDA, and maximal phase advances were seen after 100 mM NMDA. A phase-response curve (PRC) for NMDA, determined by administering NMDA at each hour over the circadian cycle, resembled the PRC to light in this species. These data support the hypothesis that NMDA-type glutamate receptors play a critical role in mediating the phase shifting effects of light in diurnal, as well as nocturnal, animals. In addition, these data suggest that diurnal grass rats may be less sensitive to the phase shifting properties of NMDA than nocturnal rodents.