Grass Rat Research Articles

Compartmentalized expression of light-induced clock genes in the suprachiasmatic nucleus of the diurnal grass rat (Arvicanthis niloticus)

Photic responses of the circadian system are mediated through light-induced clock gene expression in the suprachiasmatic nucleus (SCN). In nocturnal rodents, depending on the timing of light exposure, Per1 and Per2 gene expression shows distinct compartmentalized patterns that correspond to the behavioral responses. Whether the gene- and region-specific induction patterns are unique to nocturnal animals, or are also present in diurnal species is unknown. We explored this question by examining the light-induced Per1 and Per2 gene expression in functionally distinct SCN subregions, using diurnal grass rats Arvicanthis niloticus. Light exposure during nighttime induced Per1 and Per2 expression in the SCN, showing unique spatiotemporal profiles depending on the phase of the light exposure. After a phase delaying light pulse (LP) in the early night, strong Per1 induction was observed in the retinorecipient core region of the SCN, while strong Per2 induction was observed throughout the entire SCN. After a phase advancing LP in the late night, Per1 was first induced in the core and then extended into the whole SCN, accompanied by a weak Per2 induction. This compartmentalized expression pattern is very similar to that observed in nocturnal rodents, suggesting that the same molecular and intercellular pathways underlying acute photic responses are present in both diurnal and nocturnal species. However, after an LP in early subjective day, which induces phase advances in diurnal grass rats, but not in nocturnal rodents, we did not observe any Per1 or Per2 induction in the SCN. This result suggests that in spite of remarkable similarities in the SCN of diurnal and nocturnal rodents, unique mechanisms are involved in mediating the phase shifts of diurnal animals during the subjective day.

From daily behavior to hormonal and neurotransmitters rhythms: Comparison between diurnal and nocturnal rat species

Mammalian species can be defined as diurnal or nocturnal, depending on the temporal niche during which they are active. Even if general activity occurs during nighttime in nocturnal rodents, there is a patchwork of general activity patterns in diurnal rodents, including frequent bimodality (so-called crepuscular pattern, i.e., dawn and dusk peaks of activity) and a switch to a nocturnal pattern under certain circumstances. This raises the question of whether crepuscular species have a bimodal or diurnal – as opposed to nocturnal – physiology. To this end, we investigated several daily behavioral, hormonal and neurochemical rhythms in the diurnal Sudanian grass rat ( Arvicanthis ansorgei) and the nocturnal Long–Evans rat ( Rattus norvegicus). Daily rhythms of general activity, wheel-running activity and body temperature, with or without blocked wheel, were diurnal and bimodal for A. ansorgei, and nocturnal and unimodal for Long–Evans rats. Moreover, A. ansorgei and Long–Evans rats exposed to light–dark cycles were respectively more and less active, compared to conditions of constant darkness. In contrast to other diurnal rodents, wheel availability in A. ansorgei did not switch their general activity pattern. Daily, unimodal rhythm of plasma leptin was in phase-opposition between the two rodent species. In the hippocampus, a daily, unimodal rhythm of serotonin in A. ansorgei occurred 7 h earlier than that in Long–Evans rats, whereas a daily, unimodal rhythm of dopamine was unexpectedly concomitant in both species. Multiparameter analysis demonstrates that in spite of bimodal rhythms linked with locomotor activity, A. ansorgei have a diurnally oriented physiology.

The electroretinogram (ERG) of a diurnal cone-rich laboratory rodent, the Nile grass rat ( Arvicanthis niloticus)

The most widespread models to study blindness, rats and mice, have retinas containing less than 3% cones. The diurnal rodent Arvicanthis niloticus retina has around 35% cones. Using ERG recordings, we studied retina function in this species. Several features differed from that reported in rats and mice: (a) fivefold larger photopic a-wave amplitudes; (b) photopic hill effect in Nile grass rats only; and (c) flicker amplitude plateau between 5 to 35 Hz with fusion beyond 60 Hz in Nile grass rats only. We conclude that A. niloticus might complement rats and mice for studying retinal function and pathologies involving cones.

Breeding patterns of Arvicanthis neumanni in central Tanzania

AbstractThe breeding pattern of the grass rat, Arvicanthis neumanni, was investigated in central Tanzania in 2002/2003. Nine hundred and forty‐seven animals were captured in three age groups: juveniles, sub adults and adults. These age groups were present in the population for an extended duration, but there was an increase in the number of sexually active individuals and juveniles 2–3 months after the onset of the rains (December 2002–May 2003). A peak in reproductive activity occurred between January and April. Litter size ranged between 5.58 ± 0.42 and 6.1 ± 0.26 in two study sites. There were no significant differences in the number of embryos implanted in the right and left horns of the uterus of pregnant females (t22 = 0, P > 0.05 and t36 = 1.68, P > 0.05, respectively). Sex ratio of A. neumanni was not skewed to either males or females. Breeding was seasonal and seemed to be associated with seasonal variations in primary productivity, which relates to rainfall patterns.

Retinal anatomy and visual performance in a diurnal cone‐rich laboratory rodent, the Nile grass rat (Arvicanthis niloticus)

Unlike laboratory rats and mice, muridae of the Arvicanthis family (A. ansorgei and A. niloticus) are adapted to functioning best in daylight. To date, they have been used as experimental models mainly in studies of circadian rhythms. However, recent work aimed at optimizing photoreceptor-directed gene delivery vectors (Khani et al. [2007] Invest Ophthalmol Vis Sci 48:3954-3961) suggests their potential usefulness for studying retinal pathologies and therapies. In the present study we analyzed the retinal anatomy and visual performance of the Nile grass rat (A. niloticus) using immunohistofluorescence and the optokinetic response (OKR). We found that approximately 35-40% of photoreceptors are cones; that many neural features of the inner retina are similar to those in other diurnal mammals; and that spatial acuity, measured by the OKR, is more than two times that of the usual laboratory rodents. These observations are consistent with the known diurnal habits of this animal, and further support its pertinence as a complementary model for studies of structure, function, and pathology in cone-rich mammalian retinae.

Daily rhythms in PER1 within and beyond the suprachiasmatic nucleus of female grass rats (Arvicanthis niloticus)

Although circadian rhythms of males and females are different in a variety of ways in many species, their mechanisms have been primarily studied in males. Furthermore, rhythms are dramatically different in diurnal and nocturnal animals but have been studied predominantly in nocturnal ones. In the present study, we examined rhythms in one element of the circadian oscillator, the PER1 protein, in a variety of cell populations in brains of diurnal female grass rats. Every 4 h five adult female grass rats kept on a 12-h light/dark (LD) cycle were perfused and their brains were processed for immunohistochemical detection of PER1. Numbers of PER1-labeled cells were rhythmic not only within the suprachiasmatic nucleus (SCN), the locus of the primary circadian clock in mammals, but also in the peri-suprachiasmatic region, the oval nucleus of the bed nucleus of the stria terminalis, the central amygdala, and the nucleus accumbens. In addition, rhythms were detected within populations of neuroendocrine cells that contain tyrosine hydroxylase. The phase of the rhythm within the SCN was advanced compared with that seen previously in male grass rats. Rhythms beyond the SCN were varied and different from those seen in most nocturnal species, suggesting that signals originating in the SCN are modified by its direct and/or indirect targets in different ways in nocturnal and diurnal species.

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.

Photic and nonphotic inputs to the diurnal circadian clock

Diurnal animals occupy a different temporal niche from nocturnal animals and are consequently exposed to different amounts of light as well as different dangers. Accordingly, some variation exists in the way that diurnal animals synchronize their internal circadian clock to match the external 24-hour daily cycle. First, though the brain mechanisms underlying photic entrainment are very similar among species with different daily activity patterns, there is evidence that diurnal animals are less sensitive to photic stimuli compared to nocturnal animals. Second, stimuli other than light that synchronize rhythms (i.e. nonphotic stimuli) can also entrain and phase shift daily rhythms. Some of the rules that govern nonphotic entrainment in nocturnal animals as well as the brain mechanisms that control nonphotic influences on rhythms do not appear to apply to diurnal animals, however. Some evidence supports the idea that arousal or activity plays an important role in entraining rhythms in diurnal animals, either during the light (active) or dark (inactive) phases, though no consistent pattern is seen. GABAergic stimulation induces phase shifts during the subjective day in both diurnal and nocturnal animals. In diurnal Arvicanthis niloticus (Nile grass rats), SCN GABAA receptor activation at this time results in phase delays while in nocturnal animals phase advances are induced. It appears that the effect of GABA at this circadian phase results from the inhibition of period gene expression in both diurnal and nocturnal animals. Nonetheless, the resulting phase shifts are in opposite directions. It is not known what stimuli or behaviours ultimately induce changes in GABA activity in the SCN that result in alterations of circadian phase in diurnal grass rats. Taken together, studies such as these suggest that it may be problematic to apply the principles governing nocturnal nonphotic entrainment and its underlying mechanisms to diurnal species including humans.

Rhythms in a diurnal brain

The neural mechanisms governing circadian rhythms generate patterns of behavior and physiology that are very different in diurnal and nocturnal species. Here we review data bearing on the issue of where and how in the brain these differences might be generated. Molecular data from several species now confirm that the central circadian clock, located in the suprachiasmatic nucleus (SCN), is coupled to the light – dark cycle in the same manner in nocturnal and diurnal species, indicating that the fundamental differences arise from mechanisms coupling the clock to effector systems. Major differences in this coupling become apparent only when one steps beyond the SCN to look at brain regions that directly or indirectly receive input from it. This review focuses on our work on brain regions and cell populations to which the SCN projects in the diurnal species Arvicanthis niloticus (Nile grass rats). We have found rhythms in the numbers of cells containing cFos, or PER1, in a number of these regions, and the patterns of these rhythms are always different from those seen in nocturnal laboratory rats. In some areas these rhythms are simply inverted in the two species, but in other extra-SCN regions the phase of the rhythms in these two species differs in less extreme ways. Taken together, these data suggest that there is no single simple switch that causes some animals to be nocturnal and others to be diurnal. Rather, the differences likely emerge through a variety of mechanisms operating within and downstream of the cells to which the SCN projects.

AAV-Mediated Expression Targeting of Rod and Cone Photoreceptors with a Human Rhodopsin Kinase Promoter

Gene therapy for retinal degeneration requires well-defined promoters that drive expression in rod and cone photoreceptors. This study was undertaken to develop short, active derivatives of the human rhodopsin kinase (RK) gene promoter for targeting transgene expression in rods and cones. RK, also known as G protein-coupled receptor kinase 1 (GRK1), is a component of the light adaptation pathway expressed in rods and cones. Human RK (hRK) promoter and its concatemers or derivatives extending into the conserved 5' untranslated region (5'-UTR) were assayed for promoter activity in WERI retinoblastoma or Crx/Sp1-supplemented HEK-293 cells. The derivative displaying the highest activity was linked to a GFP reporter and packaged in a pseudotyped adenoassociated viral vector (AAV2/5). The AAV vector was tested in vivo by subretinal injections in wild-type mice, in the all-cone Nrl(-/-) mice, and in the cone-rich diurnal Nile grass rat (Arvicanthis niloticus). Control eyes received a similar AAV2/5 vector carrying a mouse rod opsin (mOps) promoter-controlled GFP reporter. The hRK promoter with the full 5' untranslated sequence (-112 to +180) was the most active in cell culture. Delivered by the AAV2/5 vector, RK promoter drove GFP expression specifically in photoreceptors. In rods, hRK promoter-mediated expression was as efficient as, but appeared more uniform than, mOps promoter-mediated expression. In cones, the hRK promoter drove expression, whereas the mOps promoter did not. The hRK promoter is active and specific for rod and cone photoreceptors. Because of its small size and proven activity in cones, it is a promoter of choice for somatic gene transfer and gene therapy targeting rods and cones.

Cholinergic projections to the suprachiasmatic nucleus and lower subparaventricular zone of diurnal and nocturnal rodents

In nocturnal species cholinergic agonists alter circadian rhythm phase when injected intraventricularly or directly into the suprachiasmatic nucleus (SCN), but the phase shifts obtained differ depending upon the site being injected. Cholinergic projections reach the SCN of nocturnal laboratory rats, however, nothing is known about these projections in diurnal rodents. The first objective of this study was to evaluate the hypothesis that cholinergic projections to the SCN are only present in nocturnal species. The second objective was to evaluate the hypothesis that the lower part of the subparaventricular zone (LSPV) is a candidate for being a site that mediates the phase shifts observed when cholinergic agonists are injected intraventricularly. These hypotheses were tested in the diurnal unstriped Nile grass rat ( Arvicanthis niloticus) and the nocturnal laboratory rat. Additionally, we evaluated if the light–dark (LD) cycle had an effect on the expression of the vesicular acetylcholine transporter (VAChT) in the SCN, LSPV, and in two control areas. Animals were kept in a 12:12 LD cycle and perfused at six time points. VAChT immunoreactivity was observed in the SCN, LSPV, and in the control areas of both species. The SCN and LSPV showed a differential distribution and density of cholinergic projections between the two species, but similar temporal patterns of VAChT expression were found across species. These results provide evidence for a differential cholinergic stimulation of the SCN between grass rats and laboratory rats that may reflect a rewiring of neural projections brought about by the adoption of a diurnal activity profile.

Tyrosine hydroxylase positive neurons and their contacts with vasoactive intestinal polypeptide-containing fibers in the hypothalamus of the diurnal murid rodent, Arvicanthis niloticus

Diurnal and nocturnal animals differ with respect to the timing of a host of behavioral and physiological events including those associated with neuroendocrine functions, but the neural bases of these differences are poorly understood. In nocturnal species, rhythms in tyrosine hydroxylase-containing (TH+) neurons in the hypothalamus appear to be responsible for rhythms in prolactin secretion. Here we investigated TH+ cells in a diurnal rodent ( Arvicanthis niloticus, the unstriped Nile grass rat), and comparing them with those of a nocturnal rodent ( Rattus norvegicus, Sprague–Dawley rat). We also examined relationships between TH+ cells and fibers containing vasoactive intestinal polypeptide (VIP) that are thought to originate from cells in the suprachiasmatic nucleus (SCN), the site of the primary circadian clock in mammals. The distribution of TH+ neurons was very similar in the two species except for a population of cells in the basal forebrain that was only present in grass rats. Fibers containing VIP appeared to contact neuroendocrine TH+ cells in both species. These data suggest that, though there may be subtle species differences, temporal information is likely to be carried along the same direct pathways from the SCN to the TH+ neurons in day- and night-active species.

A comparative analysis of the distribution of immunoreactive orexin A and B in the brains of nocturnal and diurnal rodents

BackgroundThe orexins (hypocretins) are a family of peptides found primarily in neurons in the lateral hypothalamus. Although the orexinergic system is generally thought to be the same across species, the orexins are involved in behaviors which show considerable interspecific variability. There are few direct cross-species comparisons of the distributions of cells and fibers containing these peptides. Here, we addressed the possibility that there might be important species differences by systematically examining and directly comparing the distribution of orexinergic neurons and fibers within the forebrains of species with very different patterns of sleep-wake behavior.MethodsWe compared the distribution of orexin-immunoreactive cell bodies and fibers in two nocturnal species (the lab rat, Rattus norvegicus and the golden hamster, Mesocricetus auratus) and two diurnal species (the Nile grass rat, Arvicanthis niloticus and the degu, Octodon degus). For each species, tissue from the olfactory bulbs through the brainstem was processed for immunoreactivity for orexin A and orexin B (hypocretin-1 and -2). The distribution of orexin-positive cells was noted for each species. Orexin fiber distribution and density was recorded and analyzed using a principal components factor analysis to aid in evaluating potential species differences.ResultsOrexin-positive cells were observed in the lateral hypothalamic area of each species, though there were differences with respect to distribution within this region. In addition, cells positive for orexin A but not orexin B were observed in the paraventricular nucleus of the lab rat and grass rat, and in the supraoptic nucleus of the lab rat, grass rat and hamster. Although the overall distributions of orexin A and B fibers were similar in the four species, some striking differences were noted, especially in the lateral mammillary nucleus, ventromedial hypothalamic nucleus and flocculus.ConclusionThe orexin cell and fiber distributions observed in this study were largely consistent with those described in previous studies. However, the present study shows significant species differences in the distribution of orexin cell bodies and in the density of orexin-IR fibers in some regions. Finally, we note previously undescribed populations of orexin-positive neurons outside the lateral hypothalamus in three of the four species examined.

Absence of circadian and photoperiodic conservation of energy expenditure in three rodent species

According to a traditional homeostatic view, living beings spend metabolic energy at a constant rate, just like a light bulb spends electrical energy, so that energy expenditure can be expressed in units of watts. However, research conducted during the last half-century has evinced pronounced circadian variation in physiological processes, not only demonstrating circadian rhythmicity in energy expenditure but also raising the hypothesis that energy expenditure may be regulated on a daily (circadian) basis rather than on a constant-rate (homeostatic) basis. In the present study, the hypothesis of circadian (and photoperiodic) conservation of energy expenditure was tested in three rodent species: domestic mice, Nile grass rats, and Syrian hamsters. Two correlates of energy expenditure (running-wheel activity and food intake) and a classic index (oxygen consumption) were used. Changes in energy expenditure were studied in animals maintained under light-dark cycles (LDs) with periods shorter or longer than 24 h as well as in animals maintained under 24-h LDs with short and long photophases. In none of the conditions in any of the species was evidence found in support of the hypothesis of circadian (or photoperiodic) conservation of energy expenditure. Energy expenditure was generally conserved on a homeostatic basis.

Light and GABAA 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.

Associations between behavior, hormones, and Fos responses to novelty differ in pre- and post-pubertal grass rats

As animals progress from one developmental stage to the next, the nature of the challenges they face can change in systematic ways, as do the mechanisms that enable them to deal effectively with them. Here we examined the changes in the behavioral patterns and neuroendocrine mechanisms associated with exposure to a novel environment before and after the transition from a pre- to a post-pubertal stage of development in the unstriped Nile grass rat ( Arvicanthis niloticus), a murid rodent that appears to live in family groups in the wild. We introduced juvenile (28 days old) and adult (60 days old) grass rats to a novel glass aquarium where they were kept for 1 h; controls were maintained in their home cages during this time. Animals were then killed, blood was sampled, and plasma corticosterone and testosterone were measured. Brains were collected and processed for immunohistochemical detection of Fos. Although behavior in the novel environment did not differ as a function of age, corticosterone secretion and Fos expression in a variety of stress-related brain regions were increased by the manipulation to a greater extent in the juveniles compared to the adults. The data suggest a pattern of development in which a novel environment that elicits the same levels of exploratory behaviors in young and adult animals triggers a greater response in stress-related brain regions as well as corticosterone secretion in the more vulnerable young ones.

Temporal and spatial distribution of immunoreactive PER1 and PER2 proteins in the suprachiasmatic nucleus and peri-suprachiasmatic region of the diurnal grass rat ( Arvicanthis niloticus)

The suprachiasmatic nucleus (SCN) of the hypothalamus contains the primary circadian pacemaker in both diurnal and nocturnal mammals. The lower subparaventricular zone (LSPV) immediately dorsal to the SCN may also play an important role in the regulation of circadian rhythms. The SCN contains a multitude of oscillator cells that generate circadian rhythms through transcriptional/translational feedback loops involving a set of clock genes including per1 and per2. Little is known about the temporal and spatial features of the proteins encoded by these genes in day-active mammals. The first objective of this study was to characterize the expression of PER1 and PER2 in the SCN of a diurnal rodent, the unstriped Nile grass rat ( Arvicanthis niloticus). The second objective was to evaluate the hypothesis that a molecular clock could exist in the LSPV, where endogenous rhythms in Fos expression are seen in grass rats but not in laboratory rats. Animals were kept on a 12:12 light/dark cycle and perfused at 4-h intervals, and their brains were processed for immunohistochemical detection of PER1 and PER2. Both proteins were seen in the SCN where they peaked early in the dark phase, providing further evidence that the differences between diurnal and nocturnal patterns of behavior emerge from mechanisms lying downstream from the pacemaker within the SCN. Rhythmic expression of PER1 and PER2 was also seen in the LSPV providing support for the hypothesis that this region might participate in circadian time keeping in the diurnal grass rat. In addition, rhythms were seen lateral to the LSPV and the SCN. Results of this study are discussed in light of similarities and differences in the circadian time-keeping systems of day- and night-active animals.

Variability of diurnality in laboratory rodents

The locomotor activity rhythms of domestic mice, laboratory rats, Syrian hamsters, Siberian hamsters, Mongolian gerbils, degus, and Nile grass rats were compared. Running-wheel activity was monitored under a light-dark cycle with 12 h of light and 12 h of darkness per day. Nile grass rats were found to be reliably diurnal, whereas laboratory rats, Siberian hamsters, domestic mice, and Syrian hamsters were reliably nocturnal. Both diurnal and nocturnal subgroups were observed in Mongolian gerbils and degus. A downward gradient of diurnality was observed from Mongolian gerbils classified as diurnal, degus classified as diurnal, gerbils classified as nocturnal, and degus classified as nocturnal. Nocturnal degus remained nocturnal when tested with an infrared motion detector without running wheels. Thus, although the diurnal-nocturnal dichotomy could be applied to some of the species, it was not appropriate for others. The dichotomy may reflect researchers' needs for systematization more than a natural distinction between species. Through mechanisms as yet poorly understood, the balance between entraining and masking processes seems to generate a gradient of temporal niches that runs from predominantly diurnal species to predominantly nocturnal species with many chronotypes in between, including species that exhibit wide intra-species gradients of temporal niche.

Tu-P7:107 Identifying unique protein kinase B pathways using proteome activity analysis and transcriptome mining of SMC with distinct phenotypes

The same clock-genes, including Period (PER) 1 and 2, that show rhythmic expression in the suprachiasmatic nucleus (SCN) are also rhythmically expressed in other brain regions that serve as extra-SCN oscillators. Outside the hypothalamus, the phase of these extra-SCN oscillators appears to be reversed when diurnal and nocturnal mammals are compared. Based on mRNA data, PER1 protein is expected to peak in the late night in the paraventricular nucleus of the hypothalamus (PVN) of nocturnal laboratory rats, but comparable data are not available for a diurnal species. Here we use the diurnal grass rat (Arvicanthis niloticus) to describe rhythms of PER1 and 2 proteins in the PVN of animals that either show the species-typical day-active (DA) profile, or that adopt a night-active (NA) profile when given access to running wheels. For DA animals housed with or without wheels, significant rhythms of PER1 or PER2 protein expression featured peaks in the late morning; NA animals showed patterns similar to those expected from nocturnal laboratory rats. Since the PVN is part of the circuit that controls pineal rhythms, we also measured circulating levels of melatonin during the day and night in DA animals with and without wheels and in NA wheel runners. All three groups showed elevated levels of melatonin at night, with higher levels during both the day and night being associated with the levels of activity displayed by each group. The differential phase of rhythms in the clock-gene protein in the PVN of diurnal and nocturnal animals presents a possible mechanism for explaining species differences in the phase of autonomic rhythms controlled, in part, by the PVN. The present study suggests that the phase of the oscillator of the PVN does not determine that of the melatonin rhythm in diurnal and nocturnal species or in diurnal and nocturnal chronotypes within a species.

Individual Differences in Rhythms of Behavioral Sleep and Its Neural Substrates in Nile Grass Rats

Laboratory populations of grass rats (Arvicanthis niloticus) housed with a running wheel show considerable variation in patterns of locomotor activity. At the extremes are "day-active" (DA) animals with a monophasic distribution of running throughout the light phase and "night-active" (NA) animals exhibiting a biphasic pattern with an extended peak at the beginning of the dark phase and a brief peak shortly before lights-on. Here, the authors use this intraspecific variation to explore interactions between circadian and homeostatic influences on sleep and the effects of these interactions on the activity of brain regions involved in sleep regulation. Male animals were singly housed with running wheels in a 12:12 LD cycle, videotaped for 24 h, and perfused at ZT 4 or 16. Behavioral sleep was scored from the videotapes, and brains were processed for cFos immunoreactivity (cFos-ir). Sleep duration within the light and dark phases was higher in NA and DA animals, respectively, but these groups did not differ with respect to total sleep. In both groups, sleep bouts were shortest in the light phase and longest between ZT 20 and ZT 23. In the ventrolateral preoptic area (VLPO), cFos-ir was higher at ZT 16 than at ZT 4 in DA but not NA grass rats, and it was correlated with behavioral sleep at ZT 16 but not ZT 4. In OXA neurons, cFos-ir was high at ZT 4 in DA grass rats and at ZT 16 in NA grass rats, and it was correlated with behavioral sleep at both times. In the lower subparaventricular zone (LSPV), cFos-ir was higher at ZT 16 in both DA and NA animals, and it was unrelated to behavioral sleep. Thus, patterns of cFos-ir in the LSPV and OXA neurons were most tightly linked to time and sleep, respectively, whereas cFos-ir in the VLPO was influenced by an interaction between these 2 variables.