Intergeniculate Leaflet Research Articles

The Thalamic Intergeniculate Leaflet Mediates Locomotor Activity-Induced Reversal of Phenotype in Photoperiod Nonresponsive Siberian Hamsters

The role of the intergeniculate leaflet of the thalamus (IGL) in photoperiod responsiveness was examined in a laboratory-selected line of photoperiod nonresponsive (NR) Siberian hamsters. NR hamsters fail to exhibit typical winter-type responses (i.e., gonadal regression and development of winter-type pelage) when exposed to short day lengths (e.g., 10 h of light/day). Earlier studies revealed that NR hamsters will exhibit winter-type responses when exposed to short photoperiod if they are given free access to a running wheel. The present study tested the hypothesis that this locomotor activity-induced reversal of phenotype is dependent on the IGL. Male NR hamsters underwent destruction of the IGL prior to being housed in short day lengths in cages equipped with running wheels. Activity rhythms were monitored for 8 weeks, after which time pelage response and paired testes weights were obtained. In contrast to sham-operated NR animals given access to running wheels, IGL-ablated animals showed no increase in the duration of nocturnal running wheel activity and became active later in the night than sham-lesioned animals. Lesioned animals also failed to exhibit the typical short photoperiod-induced gonadal regression and pelage molt. The results implicate the IGL in the mechanism by which running wheel activity can influence photoperiodic responses.

Central projections of melanopsin‐expressing retinal ganglion cells in the mouse

A rare type of ganglion cell in mammalian retina is directly photosensitive. These novel retinal photoreceptors express the photopigment melanopsin. They send axons directly to the suprachiasmatic nucleus (SCN), intergeniculate leaflet (IGL), and olivary pretectal nucleus (OPN), thereby contributing to photic synchronization of circadian rhythms and the pupillary light reflex. Here, we sought to characterize more fully the projections of these cells to the brain. By targeting tau-lacZ to the melanopsin gene locus in mice, ganglion cells that would normally express melanopsin were induced to express, instead, the marker enzyme beta-galactosidase. Their axons were visualized by X-gal histochemistry or anti-beta-galactosidase immunofluorescence. Established targets were confirmed, including the SCN, IGL, OPN, ventral division of the lateral geniculate nucleus (LGv), and preoptic area, but the overall projections were more widespread than previously recognized. Targets included the lateral nucleus, peri-supraoptic nucleus, and subparaventricular zone of the hypothalamus, medial amygdala, margin of the lateral habenula, posterior limitans nucleus, superior colliculus, and periaqueductal gray. There were also weak projections to the margins of the dorsal lateral geniculate nucleus. Co-staining with the cholera toxin B subunit to label all retinal afferents showed that melanopsin ganglion cells provide most of the retinal input to the SCN, IGL, and lateral habenula and much of that to the OPN, but that other ganglion cells do contribute at least some retinal input to these targets. Staining patterns after monocular enucleation revealed that the projections of these cells are overwhelmingly crossed except for the projection to the SCN, which is bilaterally symmetrical.

Intergeniculate leaflet: Contributions to photic and non-photic responsiveness of the hamster circadian system

The circadian visual system is able to integrate light energy over time, enabling phase response and Fos induction in the suprachiasmatic nucleus to increase in proportion to the total energy of the photic stimulus. In the present studies, the contribution of the intergeniculate leaflet to light energy integration by the hamster circadian rhythm system was evaluated. Fos protein is induced in intergeniculate leaflet neurons at much lower irradiance levels than seen in suprachiasmatic nucleus neurons. Bilateral N-methyl- d-aspartate lesions of the intergeniculate leaflet decreased phase response of the circadian locomotor rhythm to high irradiance and, in animals exposed to long duration light stimuli, reduced Fos induction in the suprachiasmatic nucleus. Normal photon integration, as indicated by attenuated rhythm phase shifts and Fos induction in suprachiasmatic nucleus cells in response to the energy in light stimuli, does not occur in the absence of the intergeniculate leaflet and is likely to be a property of the circadian rhythm system, rather than solely of the suprachiasmatic nucleus. Anatomical analysis showed that virtually no intergeniculate leaflet neurons projecting to the suprachiasmatic nucleus contain Fos induced by either light or locomotion in a novel wheel. However, cells projecting to the pretectum were found to contain novel-wheel induced Fos. The intergeniculate leaflet is implicated in the normal assessment of light by the circadian rhythm system, but the circuitry by which either photic or non-photic information gains access to the suprachiasmatic nucleus may be more complex than previously thought.

Potentiation of the resetting effects of light on circadian rhythms of hamsters using serotonin and neuropeptide Y receptor antagonists

Circadian rhythms are entrained by light/dark cycles. In hamsters, the effects of light on circadian rhythms can be modulated by serotonergic input to the suprachiasmatic nucleus from the raphe nuclei and by neuropeptide Y containing afferents to the suprachiasmatic nucleus from the intergeniculate leaflet in the thalamus. In this study we measured effects of compounds acting on serotonergic 1A and neuropeptide Y Y5 receptors to determine if combined serotonergic-neuropeptide Y inhibition could synergistically potentiate effects of light on rhythms. We used mixed serotonergic agonist/antagonists BMY 7378 or NAN-190 as well as a neuropeptide Y Y5 antagonist CP-760,542. Both BMY 7378 and NAN-190 are thought to block serotonin release via acting as agonists at the 5-hydroxytryptamine 1A (5-HT1A) autoreceptors on cells in the raphe, and also block response of target cells by acting as antagonists at post-synaptic 5-HT1A receptors, for example, in the suprachiasmatic nuclei or the intergeniculate leaflet. Replicating prior work, we found that pretreatment with either drug alone increased the phase shift to light at circadian time 19. The combined effect of BMY 7378 and CP-760,542 given prior to light at circadian time 19 was to further potentiate the subsequent phase shift in wheel-running rhythms (the phase shift was 317% of controls; light alone: 1.35 h phase shift vs. BMY 7378, CP-760,542, and light: 4.27 h phase shift). Combined treatment with NAN-190 and CP-760,542 produced a light-induced phase shift 576% of controls (phase shift to light alone: 1.23 h vs. NAN-190, CP-760,542, and light: 7.1 h phase shift). These results suggest that the resetting effects of light on circadian rhythms can be greatly potentiated in hamsters by using pharmacological treatments that block both serotonergic and neuropeptide Y afferents to the suprachiasmatic nuclei.

Modifications of local cerebral glucose utilization during circadian food-anticipatory activity

Food-anticipatory activity that animals express before a daily timed meal is considered as the behavioral output of a feeding-entrainable oscillator whose functional neuroanatomy is still unknown. In order to identify the possible brain areas involved in that timing mechanism, we investigated local cerebral metabolic rate for glucose during food-anticipatory activity produced either by a 4-h daily access to food starting 4 h after light onset or by a hypocaloric feeding provided at the same time. Local cerebral metabolic rate for glucose measured by the labeled 2-[ 14C]-deoxyglucose technique was quantified in 40 structures. In both groups of food-restricted rats, three brain regions (the nucleus of the solitary tract, the cerebellar cortex and the medial preoptic area) showed a decrease in local cerebral metabolic rate for glucose, compared with control ad libitum animals. In addition, only one structure, the paraventricular thalamic nucleus, was affected by temporal restricted feeding, and not by hypocaloric feeding, compared with ad libitum rats. By contrast, three brain regions, i.e. the intergeniculate leaflets, the paraventricular hypothalamic and the arcuate nuclei, showed specifically metabolic decreases during anticipation of hypocaloric feeding, and not during anticipation of temporal restricted feeding, compared with the ad libitum group. Expression of food-anticipatory activity appears to be regulated by an integrated neural circuit of brainstem and hypothalamic pathways, with hypocaloric feeding involving more extensive forebrain areas than temporal restricted feeding.

Pinealectomy Reduces Optic Nerve But Not Intergeniculate Leaflet Input to the Suprachiasmatic Nucleus at Night

The suprachiasmatic nucleus (SCN) of the hypothalamus regulates circadian rhythms in mammals. It receives, among others, direct inputs from the retina and from the thalamic intergeniculate leaflet (IGL). The former sends photic signals to the SCN, whereas the latter probably integrates photic and nonphotic information. To characterise these inputs in vivo, extracellular single-unit recordings were made from the SCN of rats under urethane anaesthesia during electrical stimulation of the optic nerve (OptN) or the IGL region. Cell responses were evaluated by creating peri-stimulus time histograms. Because humoral signals such as melatonin might modulate the activity of the SCN in addition to neural inputs, recordings were also made using pinealectomised (Px) rats to test for a possible role of this hormone in regulating inputs to the SCN. A significantly greater number of cells responded to IGL (60 of 90, 67%) than to OptN (35 of 75, 47%) stimulation in intact animals (chi(2) = 5.905, P = 0.015). The same was true when Px animals were tested (IGL, 82 of 131, 63%; OptN, 31 of 111, 28%; chi(2) = 27.637, P < 0.001). In intact animals, the proportion of cells responsive to IGL stimulation during the day and during the night was not significantly different from the proportion responsive in Px animals. The same was true for OptN stimulation during the day. However, during the night, the proportion of cells responsive to OptN stimulation in intact animals was significantly greater than the proportion responsive in Px animals (chi(2) = 7.127, P = 0.008). Our findings suggest that a lack of melatonin modulates OptN but not IGL inputs to the SCN.

A daily palatable meal without food deprivation entrains the suprachiasmatic nucleus of rats

Food is considered a potent Zeitgeber for peripheral oscillators but not for the suprachiasmatic nucleus (SCN), which is entrained principally by the light-dark cycle. However, when food attains relevant properties in quantity and quality, it can be a potent Zeitgeber even for the SCN. Here we evaluated the entrainment influence of a daily palatable meal, without regular food deprivation, on the circadian rhythm of locomotor activity and the c-Fos and PER-1 protein expression in the SCN. Rats fed ad libitum, in constant darkness, received a palatable meal for 6 weeks starting in the middle of the subjective day. Locomotor activity showed entrainment when the offset of activity coincided with the palatable meal-time. In the SCN, the peak expression of c-Fos was observed at palatable meal-time and PER-1 showed a peak during the onset of subjective night, as predicted according to the behavioural entrained pattern. In addition, c-Fos and PER-1 expression in the paraventricular thalamic nucleus (PVT) showed increased expression at palatable meal-time, while the intergeniculate leaflet did not, suggesting that the PVT may be involved as an input pathway of palatable food-entrainment to the SCN. These results demonstrate that daily access to a palatable meal can entrain the SCN; several stimuli can be implicated in this process, including motivation and arousal.

The selective tachykinin neurokinin 1 (NK 1) receptor antagonist, GR 205,171, stereospecifically inhibits light-induced phase advances of hamster circadian activity rhythms

Circadian rhythms in mammals are generated by master pacemaker cells located within the suprachiasmatic nucleus of the hypothalamus. In hamsters, the suprachiasmatic nucleus contains a small collection of cells immunoreactive for substance P, the endogenous ligand of tachykinin neurokinin 1 (NK 1) receptors. In addition, two other nuclei which form part of the circadian system, the intergeniculate leaflet of the thalamus and the raphe nuclei, also contain fibers and/or cell bodies immunoreactive for substance P. In light of these observations, we evaluated the influence of the selective tachykinin NK 1 receptor antagonist, GR 205,171, upon circadian activity rhythms in the hamster. Systemic injection of GR 205,171 dose-dependently (2.5–40.0 mg/kg, i.p.) inhibited light-induced phase advances in hamster circadian wheel running activity rhythms by approximately 50%. In contrast, GR 226,206, the less active enantiomer of GR 205,171, failed to affect light-induced phase advances. In addition, we examined the potential ability of GR 205,171 to induce non-photic phase shifts in hamster wheel running rhythms when injected at mid-day to late night circadian times. However, GR 205,171 (40 mg/kg) did not elicit non-photic phase shifts at these times indicating that tachykinin NK 1 receptor antagonists are only effective when a light stimulus is applied to the pacemaker. Although GR 205,171 may, in theory, activate several sites within the circadian system, we suggest that GR 205,171 acts in the raphe nuclei to increase inhibitory serotonergic input to pacemaker cells in the suprachiasmatic nuclei, thereby suppressing photic modulation of the pacemaker. These findings have important implications for the use of tachykinin NK 1 receptor antagonists in the treatment of depression and other central nervous system disorders.

Juxtacellular Recording/Labeling Analysis of Physiological and Anatomical Characteristics of Rat Intergeniculate Leaflet Neurons

The thalamic intergeniculate leaflet (IGL) is involved in mediating effects of both photic and nonphotic stimuli on mammalian circadian rhythms. IGL neurons containing neuropeptide Y (NPY) have been implicated in mediating nonphotic effects, but little is known about those involved in photic entrainment. We used juxtacellular recording/labeling in rats to characterize both photic responses and neurochemical phenotypes of neurons in the lateral geniculate area, focusing on the IGL and ventral lateral geniculate (VLG). Single neurons were recorded to characterize photic responsiveness and were labeled with Neurobiotin (Nb); tissue was stained for Nb, NPY, and in some cases for orexin A. Three classes of neurons were identified in the IGL/VLG. Type I neurons lacked NPY and showed sustained activations during retinal illumination and moderate firing rates in darkness. Type II neurons contained large amounts of NPY throughout the soma and showed varied responses to illumination: suppression, complex responses, or no response. Type III neurons had patches of NPY both on the external soma surface and within the soma, apparently representing internalization of NPY. Type III neurons resembled type I cells in their sustained activation by illumination but were virtually silent during the intervening dark period. These neurons appear to receive NPY input, presumably from other IGL cells, which may suppress their activity during darkness. These results demonstrate the presence of several classes of neurons in the IGL defined by their functional and anatomical features and reinforce the role of the IGL/VLG complex in integrating photic and nonphotic inputs to the circadian system.

Tissue‐specific expression of tryptophan hydroxylase mRNAs in the rat midbrain: anatomical evidence and daily profiles

Serotonin (5-HT) is involved in both photic and non-photic synchronization of the mammalian biological clock located in the suprachiasmatic nuclei (SCN). We have previously demonstrated that tryptophan hydroxylase protein (TPH), the rate-limiting enzyme of 5-HT synthesis, shows circadian rhythmicity in the pathways projecting from the raphe nuclei to the intergeniculate leaflets of the thalamus on one hand, and to the SCN on the other hand. In this study, we investigate whether the circadian rhythmicity in TPH protein could result from the rhythmic expression of tph gene in the raphe nuclei. We thus cloned specific tph1 and tph2 partial cDNAs and assessed the daily profiles of TPH mRNA levels by in situ hybridization in the rat raphe nuclei. Our results demonstrate that: (i) tph2 gene is exclusively expressed in the raphe nuclei, whereas tph1 gene is expressed in the pineal gland; (ii) under light-dark cycle (LD), TPH2 mRNA levels present daily variation within both median and dorsal raphe nuclei; (iii) under constant darkness TPH2 mRNA levels in both nuclei exhibit the same variation reported under LD cycle. These data show that the circadian 5-HT synthesis within the serotonergic neurons projecting to the circadian system might be explained by the rhythmic transcription of the tph2 gene in raphe nuclei. Taking our result with previous data into consideration, we further suggest that 5-HT synthesis and release within the circadian system could be directly or indirectly under the control of the SCN.

Orexin fibers form appositions with Fos expressing neuropeptide-Y cells in the grass rat intergeniculate leaflet

Neuropeptide-Y (NPY) cells in the intergeniculate leaflet (IGL) are known to modulate effects of arousal on the mammalian circadian system. However, the route through which this information reaches the IGL has not been established. Here, we provide evidence that the orexins (hypocretins) are uniquely positioned as a potential source of activity state feedback to the IGL in the grass rat, Arvicanthis niloticus. Specifically, many NPY cells in the grass rat IGL exhibit orexin-A (OXA) fiber appositions. Furthermore, NPY cells contacted by OXA fibers are significantly more likely to express Fos during nocturnal wheel running than are NPY cells without such contacts ( P < 0.001).

Dark pulse suppression of P‐ERK and c‐Fos in the hamster suprachiasmatic nuclei

It is well-established that light pulses regulate components of the extracellular signal-regulated kinases I/II (ERK) cascade in the suprachiasmatic nuclei (SCN) circadian clock. These events are important for photic-resetting of the circadian clock. The SCN circadian clock is also reset by pulses of dark, but it is unknown if this stimulus alters the activity of ERK, the transcription factor Elk-1 or expression of the immediate early gene c-fos in the SCN. Using Syrian hamsters free-running in constant light, we determined the effects of dark pulses on these factors in the SCN. In constant light, levels of phosphorylated ERK (P-ERK) showed significant circadian variation in the Syrian hamster SCN, while levels of c-Fos or phosphorylated Elk-1 (P-Elk-1) did not. A 6-h dark pulse beginning at circadian time (CT) 8 down-regulated expression of P-ERK and c-Fos, but not P-Elk-1, in the SCN. Following termination of the pulse, levels of c-Fos increased above time-matched control values, while P-ERK expression did not. When given at the beginning of the subjective night (CT13), a 6-h dark pulse did not phase-shift behavioural rhythms and failed to alter the expression of c-Fos, P-ERK, or P-Elk-1 in the SCN. At the level of the visual thalamus, expression of c-Fos in the intergeniculate leaflet was higher during the subjective night as compared to the subjective day, although dark pulses had no robust effects on expression of c-Fos or P-ELK-1 in this structure. We conclude that dark-pulse resetting of the circadian clock is complex and involves both non-photic and photic components.

Descending projections of the hamster intergeniculate leaflet: Relationship to the sleep/arousal and visuomotor systems

The intergeniculate leaflet (IGL), homolog of the primate pregeniculate nucleus, modulates circadian rhythms. However, its extensive anatomical connections suggest that it may regulate other systems, particularly those for visuomotor function and sleep/arousal. Here, descending IGL-efferent pathways are identified with the anterograde tracer, Phaseolus vulgaris leucoagglutinin, with projections to over 50 brain stem nuclei. Projections of the ventral lateral geniculate are similar, but more limited. Many of the nuclei with IGL afferents contribute to circuitry governing visuomotor function. These include the oculomotor, trochlear, anterior pretectal, Edinger-Westphal, and the terminal nuclei; all layers of the superior colliculus, interstitial nucleus of the medial longitudinal fasciculus, supraoculomotor periaqueductal gray, nucleus of the optic tract, the inferior olive, and raphe interpositus. Other target nuclei are known to be involved in the regulation of sleep, including the lateral dorsal and pedunculopontine tegmentum. The dorsal raphe also receives projections from the IGL and may contribute to both sleep/arousal and visuomotor function. However, the locus coeruleus and medial vestibular nucleus, which contribute to sleep and eye movement regulation and which send projections to the IGL, do not receive reciprocal projections from it. The potential involvement of the IGL with the sleep/arousal system is further buttressed by existing evidence showing IGL-efferent projections to the ventrolateral preoptic area, dorsomedial, and medial tuberal hypothalamus. In addition, the great majority of all regions receiving IGL projections also receive input from the orexin/hypocretin system, suggesting that this system contributes not only to the regulation of sleep, but to eye movement control as well.

Neurons expressing relaxin 3/INSL 7 in the nucleus incertus respond to stress

Relaxin 3/INSL 7 has recently been identified as a new member of the insulin/relaxin superfamily. Although it was reported to be dominantly expressed in the brain, its detailed distribution and function in the central nervous system are still obscure. In the present study we demonstrated that in the rat relaxin 3 was mainly expressed in neurons of the nucleus incertus (NI) of the median dorsal tegmental pons. Other relaxin 3-expressing neurons were scattered in the pontine raphe nucleus, the periaqueductal gray and dorsal area to the substantia nigra in the midbrain reticular formation. Relaxin 3-immunoreactive fibers projected particularly densely in the septum, hippocampus, lateral hypothalamus and intergeniculate leaflet of the thalamus. Ultrastructural examination revealed that relaxin 3 was localized in the dense-cored vesicles in the perikarya and was also observed in the synaptic terminals of axons. As almost all relaxin 3-containing neurons express corticotropin-releasing factor (CRF) type 1 receptor in the NI, we examined the response of relaxin 3 neurons to intracerebroventricular administration of CRF; 65% of relaxin 3 neurons expressed c-Fos 2 h after intracerebroventricular administration of 1 microg CRF. We then confirmed that c-Fos was induced in 60% of relaxin 3 neurons in the NI and the expression of relaxin 3 mRNA increased significantly in the NI after water-restraint stress. Collectively, these results suggest that relaxin 3 produced in the NI is released from nerve endings and is involved in the regulation of the stress response.

Hypothalamic and zona incerta neurons expressing hypocretin, but not melanin concentrating hormone, project to the hamster intergeniculate leaflet

The hypocretins (Hcrt; also known as orexins) and melanin-concentrating hormone comprise distinct families of neuropeptides synthesized in cells located in the lateral hypothalamus and adjacent areas. The Hcrts are thought to modulate food intake and sleep/wake patterns in mammals. Melanin-concentrating hormone has a well-documented role in energy metabolism. A moderate plexus of Hcrt immunoreactive terminals has been described in the hamster intergeniculate leaflet, part of the circadian rhythm system. This study investigated the origin of Hcrt-immunoreactive terminals in the intergeniculate leaflet and determined whether melanin-concentrating hormone neurons also project to the intergeniculate leaflet. The tracer, cholera toxin β-subunit, was injected into the intergeniculate leaflet of the golden hamster. Double-label fluorescent immunohistochemistry for cholera toxin β-subunit and Hcrt or melanin-concentrating hormone was then performed to identify retrogradely labeled cells also containing immunoreactive peptide. Most cholera toxin β-subunit-labeled cells were detected in the medial zona incerta and sub-incertal zone, with few observed in the lateral hypothalamus. Hcrt-immunoreactive cells were abundant and found largely in the lateral hypothalamus and adjacent nuclei. Melanin-concentrating hormone cells were also abundant in the medial zona incerta, in close proximity to cholera toxin β-subunit-labeled cells, but ventral to them. Cells containing both cholera toxin β-subunit and Hcrt-immunoreactive, were present in the dorsal aspect of the lateral hypothalamus. The number observed was small, ≤1% of the total number of Hcrt cells counted in the hamster. No cholera toxin β-subunit-immunoreactive cells also contained melanin-concentrating hormone and no melanin-concentrating hormone-immunoreactive processes were evident in the intergeniculate leaflet. The results show that a small number of lateral hypothalamus cells containing Hcrt-immunoreactivity project to the intergeniculate leaflet, but they are scattered rather than collected into a discrete group. At the present time there is no information regarding the function of these cells, although they may contribute to the regulation of sleep/arousal, circadian rhythmicity, or vestibulo-oculomotor function.

Involvement of the retinohypothalamic tract in the photic-like effects of the serotonin agonist quipazine in the rat

Light is the major synchronizer of the mammalian circadian pacemaker located in the suprachiasmatic nucleus. Photic information is perceived by the retina and conveyed to the suprachiasmatic nucleus either directly by the retinohypothalamic tract or indirectly by the intergeniculate leaflet and the geniculohypothalamic tract. In addition, serotonin has been shown to affect the suprachiasmatic nucleus by both direct and indirect serotonin projections from the raphe nuclei. Indeed, systemic as well as local administrations of the serotonin agonist quipazine in the region of the suprachiasmatic nucleus mimic the effects of light on the circadian system of rats, i.e. they induce phase-advances of the locomotor activity rhythm as well as c-FOS expression in the suprachiasmatic nucleus during late subjective night. The aim of this study was to localize the site(s) of action mediating those effects. Phase shifts of the locomotor activity rhythm as well as c-FOS expression in the suprachiasmatic nucleus after s.c. injection of quipazine (10 mg/kg) were assessed in Lewis rats, which had received either radio-frequency lesions of the intergeniculate leaflet or infusions of the serotonin neurotoxin 5,7-dihydroxytryptamine into the suprachiasmatic nucleus (25 μg) or bilateral enucleation. Lesions of intergeniculate leaflet and serotonin afferents to the suprachiasmatic nucleus did not reduce the photic-like effects of quipazine, whereas bilateral enucleation and the subsequent degeneration of the retinohypothalamic tract abolished both the phase-shifting and the FOS-inducing effects of quipazine. The results indicate that photic-like effects of quipazine are mediated via the retinohypothalamic tract.

Photostimulation alters c-Fos expression in the dorsal raphe nucleus

Retinal afferents to the dorsal raphe nucleus (DRN) have been described in a number of species, including Mongolian gerbils, but functional correlates of this optic pathway are unknown at present. To determine whether temporally modulated photostimulation can affect c-Fos expression in the gerbil DRN, quantitative analysis of c-Fos-immunoreactive (c-Fos-ir) neurons was conducted following 60-min exposure to pulsed (2 Hz) photostimulation at selected times over the 12:12 h light/dark cycle. For comparison, c-Fos expression was also analyzed in the subnuclei of the lateral geniculate complex and in the suprachiasmatic nucleus (SCN). In the DRN, a substantial reduction was observed in the number of c-Fos immunoreactive (c-Fos-ir) neurons during the light period and early dark period in photostimulated vs. control animals. Similar results were obtained in the intergeniculate leaflet (IGL) and ventral lateral geniculate (VLG). However, no significant changes were observed in the number of c-Fos-ir neurons in the dorsal lateral geniculate nucleus or suprachiasmatic nucleus (SCN) following photostimulation, except for an increase in the middle of the dark period. These findings indicate that photic stimulation can lead to a suppression or down-regulation of c-Fos expression in the DRN that is probably mediated via the direct retinal pathway to the DRN in this species. The similarity between c-Fos expression profiles in the DRN and IGL/VGL suggest that efferent projections from the DRN may modulate c-Fos expression to visual stimulation in these subnuclei of the lateral geniculate complex.

Neuropeptide Y does not reset the circadian clock in NPY Y2−/− mice

Mammalian circadian rhythms are modulated by neuropeptide Y (NPY), a peptide contained in the projection from the intergeniculate leaflet to the suprachiasmatic nuclei of the circadian pacemaker. NPY resets the circadian clock during the subjective day, mediating non-photic inputs. Previous studies using receptor-selective agonists have indicated that this action of NPY is mediated by the Y2 receptor in hamsters. The present study determined if NPY applied to the suprachiasmatic nuclei in the mid-subjective day can phase-advance the rhythm of spontaneous firing rate of Y2−/− mice. We observed that NPY did reset the rhythm of control mice but did not significantly shift the phase of this rhythm in the Y2−/− mice. These results provide strong evidence for the role of the Y2 receptor mediating neuropeptide Y subjective day phase-advance shifts in mice.

Neuronal distribution of melanin-concentrating hormone, cocaine- and amphetamine-regulated transcript and orexin B in the brain of the Djungarian hamster ( Phodopus sungorus)

The distribution of melanin-concentrating hormone-, cocaine- and amphetamine-regulated transcript- and orexin B-immunoreactive elements as well as their morphological relationships in selected brain structures harbouring the neuroendocrine pathways controlling energy balance and circadian rhythmicity in the Djungarian hamster ( Phodopus sungorus) were studied. Cocaine- and amphetamine-regulated transcript-(55–102)-immunoreactive perikarya co-expressed melanin-concentrating hormone-immunoreactivity in the lateral hypothalamic area, dorsomedial hypothalamic nucleus, zona incerta and posterior hypothalamic area. In addition, arcuate nucleus, hypothalamic periventricular nucleus, Edinger–Westphal nucleus, and the rostral aspect of the dorsal raphe nucleus contained cocaine- and amphetamine-regulated transcript-immunoreactive cell profiles. Orexin B-immunoreactive perikarya were distributed in the lateral hypothalamic area, dorsomedial hypothalamic nucleus and retrochiasmatic area. Cells immunoreactive for orexin B did not co-express melanin-concentrating hormone-immunoreactivity, but orexin B-immunoreactive fibers had close apposition to many melanin-concentrating hormone-immunoreactive cells. Whereas immunoreactivity for all examined peptides was absent in the suprachiasmatic nucleus, dense and large orexin B-immunoreactive fibers and to a lesser extent melanin-concentrating hormone- and cocaine- and amphetamine-regulated transcript-immunoreactive fibers of smaller size were present in the intergeniculate leaflet and raphe nucleus. These observations in Djungarian hamsters indicate that the neuronal distribution of the examined peptides is strongly conserved between species. In addition, the presence of fibers within the neuronal components of the circadian timing system suggests that they may indirectly influence circadian rhythms.

Differential vulnerability of the rat retina, suprachiasmatic nucleus and intergeniculate leaflet to malnutrition induced during brain development

We investigated in young rats the effects of malnutrition on the main structures of the circadian timing system: retina, hypothalamic suprachiasmatic nuclei (SCN), thalamic intergeniculate leaflet, retinohypothalamic- and geniculohypothalamic tracts. Control rats were born from mothers fed a commercial diet since gestation, and malnourished rats from mothers fed a multideficient diet since gestation (GLA group) or lactation (LA group). After weaning, pups received the same diet as their mothers, and were analysed at postnatal days 27, 30–33 and 60–63. Brain sections were processed to visualise in the SCN neuropeptide Y immunoreactivity and terminal labeling after intraocular tracer injections. Nissl staining was used to assess cytoarchitectonic boundaries of the SCN and cell features in retinal whole mounts. Cell counts, morphometric and densitometric analysis were performed. Compared with controls, the total retinal surface was reduced and the topographical distribution of retinal ganglion cells was altered in malnourished rats, with changes in their density. Alterations were also detected in the SCN dimensions in the GLA and LA groups at one and two postnatal months, as well as in the SCN portion occupied by the retinal input in the GLA group at days 30–33, but not in the NPY-containing geniculohypothalamic tract. The present data point to subtle changes, with a low and differential vulnerability to early malnutrition, of structures involved in circadian timing regulation. Furthermore, the present findings suggest that the altered circadian rhythmicity previously documented in malnourished rats cannot be ascribed to impaired development of the retino- and geniculohypothalamic projections to the SCN.