Orexin (hypocretin) mediates light-dependent fluctuation of hippocampal function in a diurnal rodent.

Environmental lighting conditions play a central role in cognitive function, but the underlying mechanisms remain unclear. Utilizing a diurnal rodent model, the Nile grass rat (Arvicanthis niloticus), we previously found that daytime light intensity affects hippocampal function in this species in a manner similar to its effects in humans. Compared to animals housed in a 12:12 h bright light-dark (brLD) cycle, grass rats kept in a 12:12 h dim light-dark (dimLD) cycle showed impaired spatial memory in the Morris water maze (MWM) and reduced CA1 apical dendritic spine density. The present study explored the neural substrates mediating the effects of daylight intensity on hippocampal function focusing on the hypothalamic orexin (hypocretin) system. First, animals housed in dimLD were treated with daily intranasal administration of orexin A peptide over five training days of the MWM task. Compared to vehicle controls, this treatment led to superior spatial memory accompanied by increased phosphorylation of Ca2+ /calmodulin-dependent protein kinase II α and glutamate receptor 1 within the CA1. To assess the role of hippocampal orexinergic signaling, an adeno-associated viral vector (AAV) expressing an orexin receptor 1 (OX1R) shRNA was injected into the dorsal hippocampus targeting the CA1 of animals housed in brLD. AAV-mediated knockdown of OX1R within the hippocampus resulted in deficits in MWM performance and reduced CA1 apical dendritic spine density. These results are consistent with the view that the hypothalamic orexinergic system underlies the modulatory role of daytime illumination on hippocampal function in diurnal mammals.

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.

Daytime Light Intensity Modulates Spatial Learning and Hippocampal Plasticity in Female Nile Grass Rats (Arvicanthis niloticus).

Light has pervasive effects on the physiology and behavior of mammals. Several human studies have shown that light modulates cognitive functions; however, the mechanisms responsible for the effects of light remain unclear. Our previous work using diurnal male Nile grass rats (Arvicanthis niloticus) revealed that reduced illuminance during the day leads to impairments in hippocampal-dependent spatial learning/memory, reduced CA1 dendritic spine density, and attenuated hippocampal brain-derived neurotrophic factor (BDNF) expression in males. The present study examined the impact of ambient light intensity on hippocampal functions in female grass rats and explored sex differences in behavioral and hippocampal responses. Female grass rats were housed in either a 12:12-hr bright light-dark (brLD, 1000 lx) or dim light-dark (dimLD, 50 lx) cycle for four weeks. The dimLD group showed impaired spatial memory in the Morris water maze task and reduced CA1 apical dendritic spine density, similar to prior observations in males. However, the behavioral deficits seen in females were more severe than those seen in males, with dimLD females showing no evidence of long-term retention over the 24-hour periods between training sessions. In contrast to the attenuated hippocampal BDNF expression found in dimLD males, there was no significant difference in the expression of BDNF and of its receptor TrkB between females in brLD and dimLD. The results suggest that, as seen in male grass rats, reduced illuminance during the day impairs hippocampal-dependent spatial memory and hippocampal plasticity in female diurnal grass rats, but the underlying signaling pathways responsible for the effects of light restriction may differ between the sexes.

Light as a modulator of emotion and cognition: Lessons learned from studying a diurnal rodent.

Light profoundly affects the behavior and physiology of almost all animals, including humans. One such effect in humans is that the level of illumination during the day positively contributes to affective well-being and cognitive function. However, the neural mechanisms underlying the effects of daytime light intensity on affect and cognition are poorly understood. One barrier for progress in this area is that almost all laboratory animal models studied are nocturnal. There are substantial differences in how light affects nocturnal and diurnal species, e.g., light induces sleep in nocturnal mammals but wakefulness in diurnal ones, like humans. Therefore, the mechanisms through which light modulates affect and cognition must differ between the chronotypes. To further understand the neural pathways mediating how ambient light modulates affect and cognition, our recent work has developed a diurnal rodent model, the Nile grass rat (Arvicanthis niloticus), in which daytime light intensity is chronically manipulated in grass rats housed under the same 12:12 hour light/dark cycle. This simulates lighting conditions during summer-like bright sunny days vs. winter-like dim cloudy days. Our work has revealed that chronic dim daylight intensity results in higher depression- and anxiety-like behaviors, as well as impaired spatial learning and memory. Furthermore, we have found that hypothalamic orexin is a mediator of these effects. A better understanding of how changes in daytime light intensity impinge upon the neural substrates involved in affect and cognition will lead to novel preventive and therapeutic strategies for seasonal affective disorder, as well as for non-seasonal emotional or cognitive impairments associated with light deficiency.

Distributions of GABAergic and glutamatergic neurons in the brains of a diurnal and nocturnal rodent.

Light influences the daily patterning of activity by both synchronizing internal clocks to environmental light-dark cycles and acutely modulating arousal states, a process known as masking. Masking responses are completely reversed in diurnal and nocturnal species. In nocturnal rodents, masking is mediated through a subset of intrinsically photosensitive retinal ganglion cells (ipRGCs) whose projections are similar in diurnal and nocturnal rodents. This raises the possibility that differences in responsivity to signals that these cells release might underlie chronotype differences in masking. We explored one aspect of this hypothesis by examining the distribution of excitatory and inhibitory neuronal populations in many ipRGC target areas of a diurnal species (Nile grass rat) and a nocturnal one (Norway rat). We discovered that while many of these regions were very similar in these two species, there were striking differences in the ventral lateral geniculate nucleus (vLGN; higher density of glutamate cells in Norway rats) and in the lateral habenula (LHb; GABAeric cells present in grass rats, but not Norway rats). These patterns raise the possibility that the vLGN and LHb contribute to differences in masking and/or circadian regulation of diurnal and nocturnal species.

The Cost of Activity during the Rest Phase: Animal Models and Theoretical Perspectives.

For humans, activity during the night is correlated with multiple pathologies that may reflect a lack of harmony among components of the circadian system; however, it remains difficult to identify causal links between nocturnal activity and different pathologies based on the data available from epidemiological studies. Animal models that use forced activity or timed sleep deprivation provide evidence of circadian disruptions that may be at the core of the health risks faced by human night and shift workers. One valuable insight from that work is the importance of changes in the distribution of food intake as a cause of metabolic imbalances associated with activity during the natural rest phase. Limitations of those models stem from the use of only nocturnal laboratory rodents and the fact that they do not replicate situations in which humans engage in work with high cognitive demands or engage voluntarily in nocturnal activity (i.e., human eveningness). Temporal niche switches by rodents have been observed in the wild and interpreted as adaptive responses to energetic challenges, but possible negative outcomes, similar to those associated with human eveningness, have not been systematically studied. Species in which a proportion of animals shows a switch from a day-active to a night-active (e.g., grass rats) when given access to running wheels provide a unique opportunity to model human eveningness in a diurnal rodent. In particular, the mosaic of phases of brain oscillators in night-active grass rats may provide clues about the circadian challenges faced by humans who show voluntary nocturnal wakefulness.

Light modulates hippocampal function and spatial learning in a diurnal rodent species: A study using male nile grass rat (Arvicanthis niloticus).

The effects of light on cognitive function have been well-documented in human studies, with brighter illumination improving cognitive performance in school children, healthy adults, and patients in early stages of dementia. However, the underlying neural mechanisms are not well understood. The present study examined how ambient light affects hippocampal function using the diurnal Nile grass rats (Arvicanthis niloticus) as the animal model. Grass rats were housed in either a 12:12 h bright light-dark (brLD, 1,000 lux) or dim light-dark (dimLD, 50 lux) cycle. After 4 weeks, the dimLD group showed impaired spatial memory in the Morris Water Maze (MWM) task. The impairment in their MWM performance were reversed when the dimLD group were transferred to the brLD condition for another 4 weeks. The results suggest that lighting conditions influence cognitive function of grass rats in a way similar to that observed in humans, such that bright light is beneficial over dim light for cognitive performance. In addition to the behavioral changes, grass rats in the dimLD condition exhibited reduced expression of brain-derived neurotrophic factor (BDNF) in the hippocampus, most notably in the CA1 subregion. There was also a reduction in dendritic spine density in CA1 apical dendrites in dimLD as compared to the brLD group, and the reduction was mostly in the number of mushroom and stubby spines. When dimLD animals were transferred to the brLD condition for 4 weeks, the hippocampal BDNF and dendritic spine density significantly increased. The results illustrate that not only does light intensity affect cognitive performance, but that it also impacts hippocampal structural plasticity. These studies serve as a starting point to further understand how ambient light modulates neuronal and cognitive functions in diurnal species. A mechanistic understanding of the effects of light on cognition can help to identify risk factors for cognitive decline and contribute to the development of more effective prevention and treatment of cognitive impairment in clinical populations.

Normal behavioral responses to light and darkness and the pupillary light reflex are dependent upon the olivary pretectal nucleus in the diurnal Nile grass rat.

The olivary pretectal nucleus (OPT) is a midbrain structure that receives reciprocal bilateral retinal projections, is involved in the pupillary light reflex, and connects reciprocally with the intergeniculate leaflet (IGL), a retinorecipient brain region that mediates behavioral responses to light pulses (i.e., masking) in diurnal Nile grass rats. Here, we lesioned the OPT and evaluated behavioral responses in grass rats to various lighting conditions, as well as their anxiety-like responses to light exposure. While control grass rats remained diurnal, grass rats with OPT lesions exhibited a more night-active pattern under 12h:12h light-dark (LD) conditions. However, when placed in constant darkness, OPT-lesioned grass rats became more active during their subjective day, suggesting that an exaggerated masking response to light may be responsible for the effect of OPT lesions on locomotor activity in LD. To test this hypothesis, we presented dark and light pulses to controls and grass rats with OPT lesions; controls increased their activity in response to light, whereas those with OPT lesions significantly increased activity in response to darkness. Further, when placed in a 7-h ultradian LD cycle, animals with OPT lesions were more active during darkness than controls. OPT lesions also abolished the pupillary light reflex, but did not affect anxiety-like behaviors. Finally, in animals with OPT lesions, light did not induce Fos expression in the ventrolateral geniculate nucleus, as it did in controls. Altogether, these results suggest that masking responses to light and darkness are dependent upon nuclei within the subcortical visual shell in grass rats.

Suprachiasmatic Nucleus and Subparaventricular Zone Lesions Disrupt Circadian Rhythmicity but Not Light-Induced Masking Behavior in Nile Grass Rats.

The ventral subparaventricular zone (vSPVZ) receives direct retinal input and influences the daily patterning of activity in rodents, making it a likely candidate for the mediation of acute behavioral responses to light (i.e., masking). We performed chemical lesions aimed at the vSPVZ of diurnal grass rats (Arvicanthis niloticus) using N-methyl-D,L-aspartic acid (NMA), a glutamate agonist. Following NMA lesions, we placed grass rats in various lighting conditions (e.g., 12:12 light-dark, constant dark, constant light); presented a series of light pulses at circadian times (CT) 6, 14, 18, and 22; and placed them in a 7-h ultradian cycle to assess behavioral masking. Extensive bilateral lesions of the vSPVZ disrupted the expression of circadian rhythms of activity and abolished the circadian modulation of masking responses to light, without affecting light-induced masking behavior per se. We also found that in diurnal grass rats, NMA was capable of destroying not only neurons of the vSPVZ but also those of the suprachiasmatic nucleus (SCN), even though excitotoxins have been ineffective at destroying cells within the SCN of nocturnal rodents. The vulnerability of the grass rat's SCN to NMA toxicity raises the possibility of a difference in density of receptors for glutamate between nocturnal and diurnal species. In cases in which damage extended to the SCN, masking responses to light were present and similar to those displayed by animals with damage restricted to the vSPVZ. Thus, extensive bilateral lesions of the SCN and vSPVZ disrupted the expression of circadian rhythms without affecting acute responses to light in a diurnal species. Our present and previous results suggest that retinorecipient brain areas other than the SCN or vSPVZ, such as the intergeniculate leaflet or olivary pretectal nucleus, may be responsible for the mediation of masking responses to light in the diurnal grass rat.

Acute effects of light on the brain and behavior of diurnal Arvicanthis niloticus and nocturnal Mus musculus.

Photic cues influence daily patterns of activity via two complementary mechanisms: (1) entraining the internal circadian clock and (2) directly increasing or decreasing activity, a phenomenon referred to as "masking". The direction of this masking response is dependent on the temporal niche an organism occupies, as nocturnal animals often decrease activity when exposed to light, while the opposite response is more likely to be seen in diurnal animals. Little is known about the neural mechanisms underlying these differences. Here, we examined the masking effects of light on behavior and the activation of several brain regions by that light, in diurnal Arvicanthis niloticus (Nile grass rats) and nocturnal Mus musculus (mice). Each species displayed the expected behavioral response to a 1h pulse of light presented 2h after lights-off, with the diurnal grass rats and nocturnal mice increasing and decreasing their activity, respectively. In grass rats light induced an increase in cFOS in all retinorecipient areas examined, which included the suprachiasmatic nucleus (SCN), the ventral subparaventricular zone (vSPZ), intergeniculate leaflet (IGL), lateral habenula (LH), olivary pretectal nucleus (OPT) and the dorsal lateral geniculate (DLG). In mice, light led to an increase in cFOS in one of these regions (SCN), no change in others (vSPZ, IGL and LH) and a decrease in two (OPT and DLG). In addition, light increased cFOS expression in three arousal-related brain regions (the lateral hypothalamus, dorsal raphe, and locus coeruleus) and in one sleep-promoting region (the ventrolateral preoptic area) in grass rats. In mice, light had no effect on cFOS in these four regions. Taken together, these results highlight several brain regions whose responses to light suggest that they may play a role in masking, and that the possibility that they contribute to species-specific patterns of behavioral responses to light should be explored in future.

Circadian modulation of memory and plasticity gene products in a diurnal species.

Cognition is modulated by circadian rhythms, in both nocturnal and diurnal species. Rhythms of clock gene expression occur in brain regions that are outside the master circadian oscillator of the suprachiasmatic nucleus and that control cognitive functions, perhaps by regulating the expression neural-plasticity genes such as brain derived neurotrophic factor (BDNF) and its high affinity receptor, tyrosine kinase B (TrkB). In the diurnal grass rat (Arvicanthis niloticus), the hippocampus shows rhythms of clock genes that are 180° out of phase with those of nocturnal rodents. Here, we examined the hypothesis that this reversal extends to the optimal phase for learning a hippocampal-dependent task and to the phase of hippocampal rhythms in BDNF/TrkB expression. We used the Morris water maze (MWM) to test for time of day differences in reference memory and monitored daily patterns of hippocampal BDNF/TrkB expression in grass rats. Grass rats showed superior long-term retention of the MWM, when the training and testing occurred during the day as compared to the night, at a time when nocturnal laboratory rats show superior retention; acquisition of the MWM was not affected by time of day. BDNF/TrkB expression was rhythmic in the hippocampus of grass rats, and the phase of the rhythms was reversed compared to that of nocturnal rodents. Our findings provide correlational evidence for the claim that the circadian regulation of cognition may involve rhythms of BDNF/TrkB expression in the hippocampus and that their phase may contribute to species differences in the optimal phase for learning.

Intergeniculate leaflet lesions result in differential activation of brain regions following the presentation of photic stimuli in Nile grass rats.

The intergeniculate leaflet (IGL) plays an important role in the entrainment of circadian rhythms and the mediation of acute behavioral responses to light (i.e., masking). Recently, we reported that IGL lesions in diurnal grass rats result in a reversal in masking responses to light as compared to controls. Here, we used Fos as a marker of neural activation to examine the mechanisms by which the IGL may influence this masking effect of light in grass rats. Specifically, we examined the patterns of Fos activation in retinorecipient areas and in brain regions that receive IGL inputs following 1-h light pulses given during the early night in IGL-lesioned and sham-operated grass rats. Three patterns emerged: (1) IGL lesions had no effect on the Fos response to light, (2) IGL lesions resulted in a reversal in Fos responses to light, and (3) IGL lesions resulted in a lack of a Fos response to light. Of specific interest were the suprachiasmatic nucleus (SCN) and the olivary pretectal nucleus (OPT), both of which are retinorecipient and connect reciprocally with the IGL. Light-induced Fos expression in the SCN was unaffected by IGL lesions, whereas the OPT exhibited a significant reduction in Fos expression following a light pulse in animals with IGL lesions. Altogether, our results suggest that the OPT, but not the SCN, exhibits a reversal in Fos responses to light following IGL lesions that reverse masking responses in diurnal grass rats. Our results suggest that interconnections between the IGL and downstream brain areas (e.g., OPT) may play a role in determining the direction of the behavioral response to light.

Behavioral Masking and cFos Responses to Light in Day- and Night-Active Grass Rats.

Light not only entrains the circadian system but also has acute effects on physiology and behavior, a phenomenon known as masking. Behavioral masking responses to bright light differ in diurnal and nocturnal species, such that light increases arousal in the former and decreases it in the latter. Comparisons made within a species that displays both diurnal and nocturnal patterns of behavior may provide insight into how masking differs between chronotypes and the association between mechanisms controlling masking and the circadian drive for activity. Nile grass rats (Arvicanthis niloticus) provide a useful model for studying such issues because when these animals are housed with running wheels, some run primarily during day, while others run at night. Here we compared behavioral masking responses to 2-h pulses of light and darkness given across a 12:12 light/dark cycle in day-active (DA) and night-active (NA) grass rats. Both wheel-running activity (WRA) and general activity (GA) were monitored. Light pulses at night tended to increase both WRA and GA overall in the DA grass rats, while in NA grass rats, light pulses significantly reduced WRA but had no effect on GA. Dark pulses during the day tended to decrease both WRA and GA in the DA grass rats, while in the NA grass rats, they tended to increase WRA in the early day but had no effect on GA overall. Next, we measured cFos expression within 2 brain areas potentially involved in masking, the intergeniculate leaflet (IGL) and the olivary pretectal area (OPT), of DA and NA grass rats either sacrificed on a control night or after a 1-h light pulse at ZT14. In DA grass rats, light at ZT14 induced cFos in the IGL and OPT, whereas in NA grass rats, cFos levels in both structures were high at ZT14 and were not altered by a 1-h light pulse. Overall, these results suggest that masking responses to light and darkness are dependent on the chronotype of the individual and that the responsiveness of the IGL and OPT to light may depend on or contribute to the behavioral response of these animals.

Lesions of the Intergeniculate Leaflet Lead to a Reorganization in Circadian Regulation and a Reversal in Masking Responses to Photic Stimuli in the Nile Grass Rat.

Light influences the daily patterning of behavior by entraining circadian rhythms and through its acute effects on activity levels (masking). Mechanisms of entrainment are quite similar across species, but masking can be very different. Specifically, in diurnal species, light generally increases locomotor activity (positive masking), and in nocturnal ones, it generally suppresses it (negative masking). The intergeniculate leaflet (IGL), a subdivision of the lateral geniculate complex, receives direct retinal input and is reciprocally connected with the primary circadian clock, the suprachiasmatic nucleus (SCN). Here, we evaluated the influence of the IGL on masking and the circadian system in a diurnal rodent, the Nile grass rat (Arvicanthis niloticus), by determining the effects of bilateral IGL lesions on general activity under different lighting conditions. To examine masking responses, light or dark pulses were delivered in the dark or light phase, respectively. Light pulses at Zeitgeber time (ZT) 14 increased activity in control animals but decreased it in animals with IGL lesions. Dark pulses had no effect on controls, but significantly increased activity in lesioned animals at ZT0. Lesions also significantly increased activity, primarily during the dark phase of a 12:12 light/dark cycle, and during the subjective night when animals were kept in constant conditions. Taken together, our results suggest that the IGL plays a vital role in the maintenance of both the species-typical masking responses to light, and the circadian contribution to diurnality in grass rats.

Pregnancy affects FOS rhythms in brain regions regulating sleep/wake state and body temperature in rats.

Circadian rhythms in behavior and physiology change substantially as female mammals undergo the transition from a non-pregnant to a pregnant state. Here, we examined the possibility that site-specific changes in brain regions known to regulate the sleep/wake cycle and body temperature might reflect altered rhythms in these overt functions. Specifically, we compared daily patterns of immunoreactive FOS in early pregnant and diestrous rats in the medial septum (MS), vertical and horizontal diagonal bands of Broca (VDB and HDB), perifornical lateral hypothalamus (LH), and ventrolateral, medial, and median preoptic areas (VLPO, MPA, and MnPO, respectively). In the pregnant animals, FOS expression was reduced and the daily rhythms of expression were lost or attenuated in the MS, VDB, and LH, areas known to support wakefulness, and in the MPA, a brain region that may coordinate sleep/wake patterns with temperature changes. However, despite the well-documented differences in sleep patterns between diestrous and pregnant rats, reproductive state did not affect FOS expression in the VLPO or MnPO, two brain regions in which FOS expression usually correlates with sleep. These data indicate that plasticity in sleep/wake patterns during early pregnancy may be driven by a reduction in wakefulness-promotion by the brain, rather than by an increase in sleep drive.

Site-specific changes in brain extra-SCN oscillators during early pregnancy in the rat.

The influence of circadian timekeeping systems on behavior and physiology can change substantially as female mammals undergo the transition from a nonpregnant to a pregnant state. Here, we examined the possibility that site-specific changes in extra-SCN oscillators and in local rhythms might coincide with the emergence of new patterns of temporal organization among various behavioral and physiological rhythms. Specifically, we compared daily patterns of immunoreactive FOS and PER2 in 3 brain regions of pregnant and diestrous rats. We found that, in the oval nucleus of the bed nucleus of the stria terminalis (BnST-ov), the peak of the PER2 rhythm occurred approximately 12 hours out of phase in pregnant and diestrous females. In contrast, the phase of the rhythm in FOS was the same, but pregnant rats expressed more FOS in the BnST-ov than diestrous ones. In the central amygdala (CEA) of diestrous females, PER2 expression was arrhythmic, but Fos expression was elevated at night. No rhythms were seen in this region of pregnant females, nor were any rhythms seen in the basolateral amygdala of either group. Overall, the patterns in the BnST-ov and the CEA were consistent with the hypothesis that differential changes in daily rhythms within some extra-SCN brain regions might mediate the changes in the temporal organization of several behavioral and endocrine functions that occur during the transition from a nonpregnant to a pregnant state.

Fos expression in arousal and reward areas of the brain in grass rats following induced wakefulness.

In the diurnal grass rat nocturnal voluntary wakefulness induces Fos expression in specific cellular populations of arousal and reward areas of the brain. Here, we evaluated whether involuntary wakefulness would result in similar patterns of Fos expression. We assessed this question using male grass rats that were sleep deprived for 6h by gentle stimulation (SD group), starting 2h before lights off (12:12 LD cycle). Then, we examined expression of Fos in cholinergic cells of the basal forebrain (BF), as well as in dopaminergic cells of the reward system, and compared these results to those obtained from an undisturbed control group. Different from previous results with grass rats that were voluntary awake, the BF of SD animals only showed a significant increase in Fos expression in non-cholinergic neurons of the medial septum (MS). These observations differ from reports for nocturnal rodents that are sleep deprived. Thus, our results show that voluntary and induced wakefulness have different effects on neural systems involved in wakefulness and reward, and that the effects of sleep deprivation are different across species. We also investigated whether other arousal promoting regions and circadian and stress related areas responded to sleep deprivation by changing the level of Fos expression. Among these areas, only the lateral hypothalamus (LH) and the ventro lateral preoptic area showed significant effects of sleep deprivation that dissipated after a 2h period of sleep recovery, as it was also the case for the non-cholinergic MS. In addition, we found that Fos expression in the LH was robustly associated with Fos expression in other arousal and reward areas of the brain. This is consistent with the view that the arousal system of the LH modulates neural activity of other arousal regions of the brain, as described for nocturnal rodents.

Projections of the suprachiasmatic nucleus and ventral subparaventricular zone in the Nile grass rat (Arvicanthis niloticus).

The phases of many circadian rhythms differ between diurnal and nocturnal species. However, rhythms within the hypothalamic suprachiasmatic nucleus (SCN), which contains the central circadian pacemaker, are very similar, suggesting that the mechanisms underlying phase preference lie downstream of the SCN. Rhythms in Fos expression in the ventral subparaventricular zone (vSPVZ), a major target of the SCN, differ substantially between diurnal Nile grass rats and nocturnal lab rats, raising the possibility that the vSPVZ modulates the effects of SCN signals at its targets. To understand better how and where the SCN and vSPVZ communicate circadian signals within the grass rat brain, we mapped their projections using the anterograde tracer biotinylated dextran amine (BDA). Adult female grass rats received unilateral BDA injections directed at the SCN or vSPVZ and their brains were perfusion-fixed several days later. Immunohistochemistry revealed that the distribution patterns of SCN and vSPVZ efferents were very similar. Labeled fibers originating in each region were heavily concentrated in the medial preoptic area, paraventricular thalamic nucleus, the subparaventricular zone, and the hypothalamic paraventricular and dorsomedial nuclei. BDA-labeled fibers from the SCN and vSPVZ formed appositions with orexin neurons and gonadotropin-releasing hormone neurons, two cell populations whose rhythms in Fos expression track temporally reversed patterns of locomotor and reproductive behavior, respectively, in diurnal and nocturnal rodents. These data demonstrate that projections of the SCN and vSPVZ are highly conserved in diurnal and nocturnal rodents, and the vSPVZ projections may enable it to modulate the responsiveness of target cells to signals from the SCN.

Circadian regulation of dailyrhythms in orexinergic neurons in diurnal and nocturnal rodents / Regulación circadiana de los ritmos diarios en neuronas

The work presented here focuses on the differential regulation of circadian rhythmi city by the central nervous systems of the diurnal Arvicanthis niloticus (or grass rat) and the nocturnal Rattus norvegicus (or lab rat). In grass rats, neurons expressing orexin (ORX) showed a significant daily endogenous rhythm in the expression of Fos that correlated with the rhythm in sleep and wakefulness, and was reversed when compared to that seen in lab rats. Ingrass rats ORX-positive neurons received substantial projections from vasoactive intestinal polypeptide (VIP) neurons of the suprachiasmatic nucleus (SCN). In contrast few VIP positive fibers were seen adjacent to ORX positive neurons in lab rats. This species difference suggests a direct control by the SCN on neurons expressing ORX in grass rats and a more indirect regulation in lab rats. These results are consistent with the hypothesis that differences between diurnal and nocturnal species are dueto differences in the functions of targets of the SCNsuch as the ORX neurons and the dorsomedial hypothalamus.(DMH)

El trabajo que se presenta aquí se centra en la regulación diferencial que ejerce el sistema nervioso central sobre ritmos circadianos en una especie diurna, Arvicanthis niloticus, o rata Nile grass y una especie nocturna, Rattus norvegicus, o rata de laboratorio. En Nile grass, las neuronas que expresan orexina (ORX) mostraron un ritmo endógeno diario en la expresión de Fos, ritmo que correlaciona con el ciclo de sueño y vigilia de esta especie y que es opuesto en comparación con el ritmo visto en ratas de laboratorio. En Nile grass las neuronas de ORX reciben proyecciones sustanciales desde neuronas del núcleo supraquiasmático (SCN) que expresan el péptido vasoactivo intestinal (VIP). En contraste, en ratas de laboratorio se encontraron muy pocas fibras positivas para VIP adyacentes a neuronas de ORX. Esta diferencia entre especies sugiere un control directo por parte del SCN sobre neuronas que expresan ORX en Nile grass y una regulación más indirecta en ratas de laboratorio. Estos resultados son consistentes con la hipótesis según la cual las diferencias entre especies diurnas y nocturnas se deben a diferencias en las funciones de regiones que reciben eferencias del SCN tales como las neuronas de ORX y el hipotálamos dorsomedial.(DMH)

Rhythms in expression of PER1 protein in the amygdala and bed nucleus of the stria terminalis of the diurnal grass rat (Arvicanthis niloticus).

In the diurnal rodent Arvicanthis niloticus (grass rats) the pattern of expression of the clock genes and their proteins in the suprachiasmatic nucleus (SCN) is very similar to that seen in nocturnal rodents. Rhythms in clock gene expression have been also documented in several forebrain regions outside the SCN in nocturnal Ratus norvegicus (lab rats). To investigate the neural basis for differences in the circadian systems of diurnal and nocturnal mammals, we examined PER1 expression in the oval nucleus of the bed nucleus of the stria terminalis (BNST-OV), and in the basolateral (BLA) and the central (CEA) amygdala of male grass rats kept in a 12:12 light/dark cycle. In the BNST-OV, peak levels of PER1 expression were seen early in the light phase of the cycle, 12h out of phase with what has been reported for nocturnal lab rats. In the BLA the pattern of PER1 expression featured sustained high levels during the day and low levels at night. PER1 expression in the CEA was also at its highest early in the light phase, but the effect of sampling time was not statistically significant (p<0.06). The results are consistent with the hypothesis that differences between nocturnal and diurnal species are due to differences in neural systems downstream from the SCN.