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.     

Circadian rhythms of vasopressin content in the suprachiasmatic nucleus of the rat.

The suprachiasmatic nucleus (SCN) of the anterior hypothalamus contains a circadian pacemaker in mammals. We determined the circadian profiles of arginine-vasopressin (AVP), a major peptide in the dorsomedial SCN, in rats under light-dark (LD), constant dark (DD) and constant light (LL) conditions. Under LD conditions, AVP levels in the SCN showed circadian rhythmicity with a peak at early light phase and a broad trough during the dark phase. This rhythm in the AVP contents was maintained even after 14 days of free-running under DD conditions and 3 days under LL conditions. These circadian patterns of AVP are similar to those of somatostatin, another peptide in the dorsomedial SCN. This indicates a common mode of regulation for peptides in this subfield of the SCN.     

Quantitative changes in neuronal and glial cells in the suprachiasmatic nucleus as a function of the lighting conditions during weaning

To examine whether lighting conditions during the development of the rat circadian system affect the morphology of the suprachiasmatic nucleus (SCN), three groups of rats were born and maintained until they were 24 days old under constant light (LL), constant darkness (DD) or 24-h light-dark cycles (LD, 12-h light and 12-h darkness). We applied a stereological method to study whether these conditions lead to alterations in the volume of the SCN and changes in the total number of neurons and glial cells. While lighting conditions did not induce differences in the SCN volume, the number of both neurons and glial cells did differ between groups. The DD rats showed the lowest number of neurons. Glial cells were also lower in this group than in the other two groups; however the number of glial cells in LL rats was lower than in LD rats. Moreover, females had more glial cells than males but males and females showed a similar number of neurons. These findings indicate the plasticity of the SCN in response to lighting conditions during the developmental stage.     

Ontogeny of light-induced Fos-like immunoreactivity in the hamster suprachiasmatic nucleus

Light induction at Fos within the Syrian hamster suprachiasmatic nucleus (SCN) occured first at postnatal day 4. The number of cells with light-induced Fos-like immunoreactivity (Fos-LI) per unit volume of SCN increased with age. Blinding experiments were used to demonstrate that the eye, though possessing an immature retina, appears to be necessary for light induction of Fos. In neonatal hamsters, environemntal cycles (e.g., light and darkness) may be able to reinforce the effect of maternal melatonin in synchronizing the pup's clock.     

Localization of vasoactive intestinal peptide and peptide histidine isoleucine immunoreactivity and mRNA within the rat suprachiasmatic nucleus

The distribution of vasoactive intestinal peptide (VIP) and peptide histidine isoleucine (PHI) mRNA within the suprachiasmatic nucleus (SCN) of rats was evaluated by immunocytochemistry and in situ hybridization. The pattern of VIP and PHI immunoreactivity corresponded closely to the distribution of VIP/PHI mRNA within the ventrolateral SCN. Clear hybridization signal was observed within the SCN of rats killed 5 h after light onset and in rats killed 2 h after the onset of the dark phase of the light-dark cycle. Visual examination of the grain density within the autoradiographs suggested that VIP/PHI mRNA may occur in higher concentrations shortly after the onset of darkness than 5 h after the onset of the light phase.     

Influence of environmental light-dark cycle and enucleation on activity of suprachiasmatic neurons in slice preparations

The influence of environmental light-dark cycle (LD) and bilateral enucleation on single neuronal activity in the suprachiasmatic nucleus (SCN) was examined using a hypothalamic slice preparation. Firstly, we reconfirmed previous results that the discharge rate in slices taken from animals kept on normal LD was higher during the light than during the dark period. Secondly, the day time discharge rate in the ventrolateral part of the SCN was decreased by bilateral enucleation and DD housing, while in the dorsomedial part it was unaffected. Thirdly, LL housing suppressed the discharge rates in both parts during the day and night periods. The present results suggest that the dorsomedial part of the SCN is more important in regulation of the circadian rhythm of SCN neuronal activity than the ventrolateral.     

Spatio-temporal analysis of light-induced Fos expression in the retina of rd mutant mice.

Rd mutant mice are visually blind but they maintain the ability of synchronising their circadian rhythms to the external light-dark cycles. We used immunocytochemical procedures to detect light-induced Fos expression in the rd mice retina. We found that Fos is expressed in the rd retina in an unattenuated pattern through the entire life of the animal. Furthermore, we have found that cells expressing Fos are distributed throughout the whole retina, while opsin expression takes place only in the dorsal half of the retina in the 1-year old rd mice. Finally, we found that light induces Fos expression in the rd retina at the same levels during the subjective day as during the subjective night, whereas in the suprachiasmatic nucleus (SCN), Fos is stimulated by light only during the subjective night. Our results support the hypothesis that new, undiscovered photoreceptors are implicated in light perception for the circadian system.     

The Role of N-Methyl-D-Aspartate-Type Glutamatergic Neurotransmission in the Photic Induction of Immediate-Early Gene Expression in the Suprachiasmatic Nuclei of the Syrian Hamster

This study investigated the role of N-methyl-D-aspartate (NMDA)-type glutamatergic neurotransmission in mediating the photic induction of immediate-early gene expression in the Suprachiasmatic nucleus (SCN) of the Syrian hamster. Activation of c-fos, c-jun and egr-1 was assessed by immunocytochemical detection of their protein products. To characterize the circadian basis to the inductive effects of light, hamsters were allowed to free-run in constant dim red light and received a 1 h light pulse at different circadian phases relative to activity onset (defined as CT 12). In control animals which did not receive light pulses, c-fos and egr-1 expression was absent or restricted to a small area of the dorsolateral region of the SCN, and expression of c-jun could not be detected in the SCN. In hamsters killed after presentation of a light pulse at either CT 14 or CT 20, there was a marked increase in c-fos and egr-1 immunoreactivities throughout the ventrolateral division of the SCN. In contrast, light pulses given at CT4 or CT 8 failed to activate immediate-early gene expression. Light pulses did not induce c-jun immunoreactivity at any circadian phase tested. Staining for c-fos was maximal 1 h after the start of the light pulse and had started to decline by 2 h. At this later time, c-jun expression was still undetectable. To compare the distribution of retinal afferents with that of c-fos induction, hamsters held on a light schedule of 16 h light: 8 h dark received an intraocular injection of cholera toxin-horseradish peroxidase conjugate 3 days before exposure to a 1 h light pulse given 2 h after lights off. Comparison of adjacent sections processed for c-fos immunoreactivity or for cholera toxin-horseradish peroxidase revealed that light-induced c-fos expression was precisely restricted to retinal terminal fields in the SCN. Light pulses also induced c-fos expression in the retinoreceptive ventral lateral geniculate nucleus and intergeniculate leaflet but not in the retinal fields of the dorsal lateral geniculate nucleus, indicating that the expression of cfos in response to light is spatially specific. The aim of the subsequent experiments was to investigate the role of NMDA-type glutamatergic neurotransmission in mediating the effects of light on c-fos expression in the SCN. To determine whether NMDA had the potential to activate c-fos expression in the SCN, hamsters were infused with 2.5 nmol NMDA or vehicle via an intracerebroventricular (icv) cannula positioned adjacent to the nuclei. In contrast to the effects of light, icv NMDA activated c-fos expression at both CT8 and CT 14. The distribution of immunoreactivity was more widespread than that observed after light, extending throughout the SCN and adjacent hypothalamus. To test whether NMDA receptors had a physiological role in the photic response, hamsters were treated systemically with the non-competitive NMDA antagonist MK801 (dose range 0.6 to 6.0 mg/kg body wt, ip) or vehicle prior to exposure to a 1 h light pulse given at CT 14 or CT 20. Expression of c-fos was still detectable in the dorsolateral SCN but MK801 blocked expression in the ventral portion of the retinoreceptive zone of the SCN. MK801 (10 or 100 nmol) delivered centrally (icv) also prevented light-induced c-fos expression in the ventral region of the SCN bordering the optic chiasm, though staining again persisted in the dorsolateral region. The induction of c-fos by icv NMDA, and the partial blockade of light-induced c-fos expression by the antagonist MK801, are consistent with the hypothesis that glutamate mediates the effects of light on SCN activity. However, the persistent photic induction of c-fos expression in a subfield of retinal afferents following treatment with MK801 suggests that other, non-NMDA-type mechanisms may contribute to photic entrainment.     

Nitric oxide synthase inhibitor blocks light-induced phase shifts of the circadian activity rhythm, but not c-fos expression in the suprachiasmatic nucleus of the Syrian hamster

Circadian rhythms in mammals are entrained to the environmental light cycle by daily adjustments in the phase of the circadian pacemaker located in the suprachiasmatic nuclei (SCN) of the hypothalamus. Brief exposure of hamsters maintained under constant darkness to ambient light during subjective nighttime produces both phase shifts of the circadian activity rhythm and characteristic patterns of c-fos protein (Fos) immunoreactivity in the SCN. In this study, we demonstrate that light-induced phase shifts of the circadian activity rhythm are blocked by intracerebroventricular (i.c.v.) injection of the competitive nitric oxide synthase (NOS) inhibitor,N-nitro-l-arginine methyl ester (l-NAME), but not by the inactive isomer,d-NAME. The effects ofl-NAME are reversible and dose-related, and are countered by co-injection of arginine, the natural substrate for NOS. While effects on behavioral rhythms are pronounced, similar treatment does not alter the pattern of light-induced Fos immunoreactivity in the SCN. These results suggest that nitric oxide is a component of the signal transduction pathway that communicates photic information to the SCN circadian pacemaker, and that nitric oxide production is either independent of, or downstream from, pathways involved in induction of c-fos expression.     

Circadian photic regulation of melatonin receptor density in rat suprachiasmatic nuclei: Comparison with light induction of fos-related protein

High-affinity melatonin receptors are present in rat suprachiasmatic nuclei (SCN), and their density exhibits a daily rhythm regulated by the light/dark cycle. In this study we demonstrate that the light regulation of these receptors depends on a circadian mechanism. Pinealectomized rats kept in constant darkness were subjected to 1-hr light pulses delivered across the circadian cycle. The density of melatonin receptors was significantly increased when photic exposure was performed during subjective night, and not different from control animals kept in darkness when the light pulse was applied during subjective day. The protein product (Fos) of the immediate early gene c-fos studied in the same paradigm showed globally the same circadian sensitivity phase. These results clearly show that, although the rhythmic appearance of melatonin receptor density in SCN follows and is directly regulated by the standard light/dark cycle, this light regulation is not passive. As is the case with Fos-like protein, it is only during a precise phase of the circadian cycle that light is able to regulate the density of melatonin receptors in SCN. © 1996 Wiley-Liss,Inc.     

Non-photic manipulations induce expression of Fos protein in the suprachiasmatic nucleus and intergeniculate leaflet in the rat

Expression of Fos protein in the suprachiasmatic nucleus (SCN) and intergeniculate leaflet (IGL) is considered a cellular correlate of light-induced phase-shift of circadian rhythms in rodents. Non-photic stimuli also induce phase shifts, but their effects on Fos expression have not been established. We examined induction of Fos protein in SCN and IGL regions, in response to cage change, intraperitoneal saline injection, and restraint stress. Fos immunoreactivity was observed in SCN and IGL regions, with greater expression observed in IGL during the light phase of the light-dark cycle. Results suggest that cells in SCN and IGL respond to several types of non-photic manipulations and that expression of Fos in these regions is not light-specific.     

Effect of NMDA Receptor Antagonist MK-801 on Light-Induced Fos Expression in the Suprachiasmatic Nuclei and on Melatonin Production in the Syrian Hamster

In mammals, circadian rhythms generated by the suprachiasmatic nuclei (SCN) are daily synchronized by a light-dark cycle. Photic information is transmitted to the SCN mainly through the direct retinohypothalamic tract, the neurotransmitters involved being excitatory amino acids. It is also commonly accepted that photoperiodic information coming from the retina via the SCN is transduced by the pineal into a nocturnal signal, i.e. melatonin production. Light exposure at night induces (1) an inhibition of melatonin synthesis and (2) an expression of c- fos in numerous cells of SCN. To determine the role of the NMDA receptor in these effects, we treated Syrian hamsters with ip injections of MK-801, a noncompetitive NMDA receptor antagonist. Several subpopulations of light-sensitive cells in the SCN are affected by MK-801. According to previous studies, MK-801 inhibits light-induced Fos immunoreactivity mainly in the most ventral part of the SCN. However, we observed that numerous other cells are still activated by light. When light is applied in the middle of the night, MK-801 pretreatment does not reduce Fos-ir in the dorsal SCN. At the beginning of the night, labeled cells in this part of the nucleus appear even more numerous after MK-801. We also found that MK-801 fails to reduce the light-induced inhibition of melatonin synthesis. Moreover, in control animals, which received no light stimulation, ip injection of MK-801 induces by itself a dose-dependent inhibition of melatonin production.     

Involvement of glial fibrillary acidic protein (GFAP) expressed in astroglial cells in circadian rhythm under constant lighting conditions in mice

To clarify the role of glial fibrillary acidic protein (GFAP)-expressed glial cells in the circadian clock, we examined GFAP expression in the suprachiasmatic nucleus (SCN) and the intergeniculate leaflet (IGL) under various lighting conditions in mice. We demonstrated that GFAP expression did not show daily change in the SCN under a light-dark cycle; however, long-term housing under constant lighting conditions led to dramatic changes in GFAP expression, i.e., a decrease in the SCN and an increase in the IGL. Furthermore, mice that had a targeted deletion in the GFAP gene (GFAP mutant mice) showed longer and more arrhythmic circadian activity rhythms in constant lighting conditions than wild-type mice, while GFAP mutant mice exhibited stable circadian rhythms both in a light-dark cycle and constant darkness, and showed normal entrainment to environmental light stimuli. These results suggest that the GFAP-expressed astroglial cells in the SCN and the IGL may have some role in circadian oscillation under constant lighting conditions.     

Effect of photic stimuli disturbing overt circadian rhythms on the dorsomedial and ventrolateral SCN rhythmicity

To ascertain how photic stimuli disturbing overt circadian rhythms affect the endogenous rhythmicity of the suprachiasmatic nucleus (SCN), rats were subjected to constant light (LL) or to a 9-h light pulse encompassing midnight, and rhythms of abundance of the c-Fos-immunoreactive (c-Fos-ir) and the PER1-immunoreactive (PER1-ir) cells were studied during the first 1-2 cycles following release into LL or darkness (DD) within the whole SCN as well as in its ventrolateral (vl) and the dorsomedial (dm) part. LL seemingly abolished the c-Fos rhythm in the whole SCN, while the rhythm persisted in the vl- and dm-SCN. In the dm-SCN, the rhythm of c-Fos-ir was phase-delayed by about 4 h in LL, whereas the rhythm of PER1-ir was affected just slightly. In the vl-SCN, the rhythm of c-Fos photo-induction might be delayed by 5-6 h as compared with the reported rhythm [A. Sumova and H. Illnerova, Am. J. Physiol. 274 (1998) R857-R863], whereas the PER1 profile appeared to be out of phase with that in DD. After a 9-h light pulse encompassing midnight, the rhythm of PER1-ir in the dm-SCN changed just slightly, whereas the PER1 rhythm in the vl-SCN was abolished and there was just an indication of extension of elevated PER1-ir. Altogether, the data indicate that photic stimuli disturbing circadian rhythms affect more dramatically the vl- than the dm-SCN rhythmicity within the first cycles and that in the dm-SCN shifting of the c-Fos rhythm proceeds more rapidly than that of the Per1 rhythm.     

Diurnal and circadian changes of serotonin in the suprachiasmatic nuclei: regulation by light and an endogenous pacemaker

Daily variations of serotonin (5-HT) in the suprachiasmatic nuclei (SNC) were measured in rats kept under various lighting conditions to elucidate the serotonergic contribution to the mechanism underlying SCN function on circadian rhythmicity. Animals kept in 12-h light-12-h dark (LD) cycles showed a peak 5-HT level during the light period and a trough during the dark period. In constant darkness (DD), rhythmic 5-HT variation was out of phase to changes observed in LD. Rats that have been kept in DD and then exposed to constant light (LL) showed transitory increases in 5-HT just after lights on. Taken together, these results show that 5-HT variation in the SCN is generated by an endogenous pacemaker and is also influenced by photic cues.     

Different populations of cells in the suprachiasmatic nuclei express c-fos in association with light-induced phase delays and advances of the free-running activity rhythm in hamsters

Circadian rhythmicity is controlled by a light-entrainable pacemaker located in the suprachiasmatic nuclei (SCN) of the mammalian hypothalamus. Brief light exposure during the subjective night causes phase shifts of the free-running activity rhythm and expression of c-fos-related proteins (Fos) among a population of cells in the hamster SCN. Light exposure (30 lux for 15 min) during the early subjective night (CT13) causes phase delays (-60 +/- 12 min), while exposure at mid-subjective night (CT18) causes phase advances (114 +/- 48 min) of the free-running activity rhythm. Light exposure at mid-subjective day (CT6) does not cause phase alterations of the rhythm. Similarly, only light exposure at CT13 or CT18 induces Fos expression in the SCN. The distribution of Fos-immunoreactive cells in the SCN is more widespread in animals stimulated with light at CT18. In addition, a group of cells located dorsal and anterior to the SCN express Fos only after stimulation at CT18. The data are consistent with the hypothesis that Fos expression represents an event in the signal transduction pathway leading to light-induced alterations in circadian pacemaker function. Furthermore, the data raise the possibility that different populations of cells in the suprachiasmatic hypothalamus may participate in light-induced phase advances and delays of the circadian pacemaker.     

Hypothalamic expression of a novel gene product, VGF: immunocytochemical analysis

VGF is the designation for a new 712 amino acid protein, regulated by nerve growth factor (NGF) in PC12 cells, that has not been previously described in the CNS. Northern blot analysis with a nick-translated VGF cDNA probe revealed a single band of mRNA in the brain with a molecular weight identical to that found in PC12 cells. The current paper presents a series of immunocytochemical studies of VGF expression with a focus on the hypothalamus. Two different antisera were raised against nonoverlapping amino acid sequences of a bacterial-expressed protein from the VGF gene cloned from PC12 cells. VGF immunoreactivity is strongly expressed in the rat suprachiasmatic nucleus (SCN), particularly in the dorsomedial part of the nucleus. The administration of colchicine to block axonal transport facilitates detection of the VGF immunoreactivity also in the ventrolateral suprachiasmatic nucleus. This protein appears to be the first one of limited neuronal distribution which is found in both dorsomedial SCN and ventrolateral SCN. Immunostaining of serial 1 micron SCN sections reveals co-localization of VGF in cells which also contain vasopressin or vasoactive intestinal polypeptide. Weaker immunoreactivity is also found in the magnocellular paraventricular and supraoptic nuclei, where the VGF immunoreactivity co-localizes with oxytocin or vasopressin. Mutant Brattleboro rats which do not express vasopressin showed strong VGF immunoreactivity both in the dorsomedial SCN and in cells of the magnocellular neuronal systems, including cells which normally express vasopressin. When axonal transport of the protein is blocked by colchicine, VGF-immunoreactive cells in the hypothalamic arcuate, parvocellular paraventricular, and tuberomammillary nuclei can also be detected, in addition to weakly immunoreactive scattered cells in the hippocampus, amygdala, thalamus, and cortex. VGF immunoreactivity is strong in the axonal projections of SCN and weak in the axons of the paraventricular and supraoptic nuclei. With ultrastructural studies, VGF immunoreactivity is found in presynaptic boutons in the SCN and in axons in the neurohypophysis. Weak axonal staining is present in some regions of the hypothalamus and in the external and internal zones of the median eminence. Immunoreactivity is absent from the intermediate lobe of the hypophysis. In neonatal rats strong VGF immunoreactivity is found throughout the SCN at postnatal day 4 but not in the adjacent hypothalamus. VGF immunoreactivity is also seen in other areas of the brain in neonatal rats, including the lateral geniculate nucleus; while the staining in the dorsal lateral geniculate disappears in the adult, that in the intergeniculate leaflet, a visual center which projects to the SCN, remains.(ABSTRACT TRUNCATED AT 400 WORDS)     

Temporospatial characteristics of light-induced Fos immunoreactivity in suprachiasmatic nuclei are not modified in Syrian hamsters treated neonatally with monosodium glutamate

Neonatal treatment of rodents by intraperitoneal injections of monosodium glutamate (MSG) destroys many retinal ganglion cells whose neurons belong to the circadian system; howertheless, adults always synchronize their locomotor activity rhythm (LAR) to the light/dark cycle. Recent studies have shown that light-induced phase shifts of LAR are associated with the c-fos induction in suprachiasmatic nuclei (SCN) of nocturnal rodents. In this study, the circadian system was analyzed in treated and control hamsters maintained in constant darkness and exposed to light at circadian times (CTs) 13 and 18 during subjective night, 1 and 6 h after the onset of LAR. The period of the LAR and delay (CT13) and advance (CT18) phase shifts of LAR were not significantly different between MSG-treated and control hamsters. Temporospatial variations of Fos induction after light exposure were similar in both MSG-treated and control hamsters although the total number of Fos immunoreactive (Fos-ir) nuclei in the SCN was always lower in treated hamsters. However, the decrease in Fos-ir was significant only for the caudal third of the SCN of treated hamsters, the part where retinal afferents are most dense. The effect of light exposure on Fos expression in SCN of MSG-treated and control hamsters was the same at CT13 and CT18: (1) Fos-ir nuclei were significantly more numerous at CT18 than at CT13 in the rostral SCN; (2) dorsal Fos-ir cells were observed in the SCN only at CT18; (3) a ventral subgroup expressed Fos protein in intermediate SCN only at CT13. This study demonstrates that MSG-treatment does not significantly modify the phase-shifting effects of light on either the LAR or the associated cellular activation.     

Serotonergic Modulation of Photically Induced Increase in Melatonin Receptor Density and Fos Immunoreactivity in the Suprachiasmatic Nuclei of the Rat

The mammalian suprachiasmatic nuclei (SCN) contain a circadian clock which is regulated by neuronal photic and non-photic afferences. Among these, the serotonergic input originating from the dorsal raphe nucleus (DRN) is extremely important. In rats, a light pulse administered during the dark period is known to induce the expression of the immediate early gene c-fos and to increase melatonin receptor density in the SCN. The aim of this study was to assess whether, in rats, these two phenomena were regulated by serotonin, acting via 5-HT_1A receptors. Three days after pinealectomy, 4 groups of rats were injected i.p.  90  min before sacrifice with respectively: (1) vehicle, (2) the 5-HT_1A-agonist 8-OH-DPAT (5  mg/kg), (3) the 5-HT_1A-antagonist NAN-190 (10  mg/kg) or (4) NAN-190 and then 8-OH-DPAT. Half of the animals from each group were exposed to light for 60  min before sacrifice and the other half remained in darkness. Sacrifice took place 5 to 6  h after lights off. Our results show that the antagonist NAN-190: (1) completely blocked the photically-induced increase of melatonin receptor density in the SCN, with an IC50=0.352±0.103  mg/kg, and (2) partially blocked (30%) the photic induction of Fos (the protein product of c-fos) in the ventrolateral subdivision of the SCN. The agonist 8-OH-DPAT enhanced the photically-induced increase of melatonin receptors by 10% and decreased the photically-induced increase in Fos by 18%. Both drugs were devoid of any effect in non-light-exposed animals. From these results we may suggest that, in rats, there is a serotonergic control of the neuronal path driving photic information to the SCN. This regulation seems to occur through 5-HT_1A or 5-HT_1A-like receptors.