Nucleus Reuniens Research Articles

Evolutionary significance of different neurochemical organisation of the internal and external regions of auditory centres in the reptilian brain: an immunocytochemical and reduced NADPH-diaphorase histochemical study in turtles

An immunocytochemical and histochemical study was undertaken of the torus semicircularis and nucleus reuniens, the mesencephalic and diencephalic auditory centres, in two chelonian species, Testudo horsfieldi and Emys orbicularis. The nucleus centralis of the torus semicircularis receives few 5-HT-, TH-, substance P-, and menkephalin-immunoreactive fibres and terminals, in marked contrast to the external nucleus laminaris of the torus semicircularis, in which 5-HT-, TH-, substance P-, and menkephalin-immunoreactive elements and cell bodies show a laminar distribution. Dense NPY-positive terminal-like profiles and cell bodies were observed in both the nuclei centralis and laminaris, and many NADPH-d-positive cell bodies were observed in the cell layers of the latter. In the nucleus reuniens, the distribution of 5-HT-, TH-, substance P-, and menkephalin-immunolabelling resembles that seen in the torus semicircularis, but at a lower density. The dorsorostral regions of the nucleus reuniens, as in the nucleus centralis, is insignificantly labelled, in contrast to the ventrocaudal regions in which labelled elements abound. NPY-positive elements are uniformly distributed throughout the nucleus, but no labelled cell bodies were observed. NADPH-d-positive fibres and terminals were observed in both dorsal and ventral regions of the nucleus reuniens, but the few labelled cell bodies to be observed were located in the peripheral regions of the nucleus. These findings are discussed in terms of the evolution of the core-and-belt organisation of sensory nuclei observed in other vertebrate species.

Suprachiasmatic nucleus projections to the paraventricular thalamic nucleus of the rat

The suprachiasmatic nucleus (SCN) projections to the midline and intralaminar thalamic nuclei were examined in the rat. Stereotaxic injections of the retrograde tracer cholera toxin-β subunit (CTb) were made in 12 different thalamic sites. These included individual midline thalamic nuclei (anterior, middle, and posterior portions of the paraventricular thalamic nucleus (PVT), intermediodorsal, paratenial, rhomboid, or reuniens nuclei) and intralaminar thalamic nuclei (lateral parafascicular, central lateral, or central medial nuclei) as well as the mediodorsal and anteroventral thalamic nuclei. After 10–14 days survival, the brains from these animals were processed histochemically and the distribution of retrogradely-labeled neurons was mapped throughout the rostralcaudal extent of the SCN. Within this collective group of midline and intralaminar thalamic nuclei, the only region innervated by the SCN was the PVT. Approximately 80% of this projection arose from the dorsomedial SCN, and the remaining projection originated from the ventrolateral SCN which targeted mainly the anterior division of the PVT. Virtually no SCN neurons were labeled after CTb injections centered in any of the other midline thalamic nuclei, which includes the intermediodorsal, mediodorsal, paratenial, rhomboid, or reuniens thalamic nuclei. Similarly, no evidence for a SCN projection to the intralaminar thalamic nuclei was found. The discussion focuses on the role of SCN → PVT pathway in modulating cerebral cortical functions.

Suprachiasmatic nucleus projections to the paraventricular thalamic nucleus of the rat

The suprachiasmatic nucleus (SCN) projections to the midline and intralaminar thalamic nuclei were examined in the rat. Stereotaxic injections of the retrograde tracer cholera toxin-β subunit (CTb) were made in 12 different thalamic sites. These included individual midline thalamic nuclei (anterior, middle, and posterior portions of the paraventricular thalamic nucleus (PVT), intermediodorsal, paratenial, rhomboid, or reuniens nuclei) and intralaminar thalamic nuclei (lateral parafascicular, central lateral, or central medial nuclei) as well as the mediodorsal and anteroventral thalamic nuclei. After 10–14 days survival, the brains from these animals were processed histochemically and the distribution of retrogradely-labeled neurons was mapped throughout the rostralcaudal extent of the SCN. Within this collective group of midline and intralaminar thalamic nuclei, the only region innervated by the SCN was the PVT. Approximately 80% of this projection arose from the dorsomedial SCN, and the remaining projection originated from the ventrolateral SCN which targeted mainly the anterior division of the PVT. Virtually no SCN neurons were labeled after CTb injections centered in any of the other midline thalamic nuclei, which includes the intermediodorsal, mediodorsal, paratenial, rhomboid, or reuniens thalamic nuclei. Similarly, no evidence for a SCN projection to the intralaminar thalamic nuclei was found. The discussion focuses on the role of SCN→PVT pathway in modulating cerebral cortical functions.

Distribution of an orphan G-protein coupled receptor (JP05) mRNA in the human brain

JP05, also called GPR72 or GIR, is an orphan G-protein-coupled receptor, GPCR, showing significant structural similarity to the tachykinin receptors. The anatomical distribution of JP05 mRNA was first described in the central nervous system of the mouse, and recently the human JP05 orphan receptor gene has been cloned. In the present study the distribution of JP05 mRNA was examined in the human forebrain using in situ hybridization analysis. The results revealed a wide but discrete distribution of the transcript with strongly JP05 mRNA expressing cells, presumably neurons, present in the cerebral cortex (layer II), hippocampus (pyramidal CA3 neurons and granule cells), amygdala (basal and periamygdaloid cortical nuclei), in the endopiriform nucleus, diagonal band of Broca, thalamus (nucleus reuniens, parafascicular nucleus) and hypothalamus (posterior, dorsal, and around the medial mammillary). Weaker signals were detected in the deeper cortical layers and throughout the striatum. A few positive cells were evident in the raphe but not in the substantia nigra or pontine nuclei. The results indicate significant similarities between human and mouse brain with regard to JP05 mRNA expression. The distribution patterns of JP05 mRNA in the human brain suggest involvement in control of emotions and of neuroendocrine, cognitive and motor functions.

Photoresponsive Fischer 344 Rats are reproductively inhibited by melatonin and differ in 2-[125I] lodomelatonin binding from nonphotoresponsive Sprague-Dawley rats.

Many temperate-zone species use photoperiod as an environmental cue to regulate reproductive timing. Strains of laboratory rats differ in their responsiveness to photoperiod, with the Fischer 344 (F344) strain being the most responsive known. F344 rats and closely related strains that differ in photoresponsiveness may be useful models to study the mechanisms and genetic basis for photoresponsiveness. We tested two hypotheses: (i) that melatonin mediates photoresponsiveness in F344 rats, as is the case in all other mammals tested, and (ii) that the location, abundance, or affinity of melatonin receptors, as estimated by the amount and location of binding of the radioligand 2-[125I]-iodomelatonin (IMEL) in the brain, might cause variation in photoresponsiveness among rat strains. Melatonin injections 1 h before lights off in a stimulatory photoperiod (L14 : D10) induced reproductive inhibition and reduced weight gain in a manner similar to short days of L8 : D16, while injections of ethanolic saline vehicle did not. Interestingly, melatonin injections administered during an inhibitory photoperiod (L10 : D14) caused greater inhibition of both reproduction and weight gain than short photoperiod alone. Pinealectomized F344 rats implanted subcutaneously with melatonin in a silastic capsule did not differ in testis size or body weight from controls with blank implants. The brains and pars tuberalis of the pituitary from photoresponsive F344 rats and nonphotoresponsive Harlan Sprague-Dawley (HSD) rats were processed for autoradiography using IMEL. We found significantly higher specific IMEL binding in the anterior and posterior regions of the paraventricular nucleus of the thalamus (PVNt) and reuniens nucleus of the thalamus of F344 rats than in the same areas in HSD rats. There were no differences between strains in specific IMEL binding in the medial PVNt, anteroventral and anterodorsal nucleus of the thalamus, suprachiasmatic nucleus, or the pars tuberalis. These results indicate that melatonin mediates photoresponsiveness in F344 rats. In addition, they provide support for the hypothesis that F344 rats may be photoresponsive due to differences from other strains in the location, density, or affinity of melatonin receptors.

Thalamic distribution of zinc-rich terminal fields and neurons of origin in the rat

Several cortico-cortical and limbic-related circuits are enriched in zinc, which is considered as an important modulator of glutamatergic transmission. While heavy metals have been detected in the thalamus, the specific presence of zinc has not been examined in this region. We have used two highly sensitive variations of the Timm method to study the zinc-rich innervation in the rat thalamus, which was compared to the distribution of acetylcholinesterase activity. The origin of some of these zinc-rich projections was also investigated by means of retrograde transport after intracerebral infusions of sodium selenium (Na 2SeO 3). The overall zinc staining in the thalamus was much lower than in the neocortex, striatum or basal forebrain; however, densely stained terminal fields were observed in the dorsal tip of the reticular thalamic nucleus, the anterodorsal and lateral dorsal thalamic nuclei and the zona incerta. In addition, moderately stained zinc-rich terminal fields were found in the rostral intralaminar nuclei, nucleus reuniens and lateral habenula. Intracerebral infusions of Na 2SeO 3 in the lateral dorsal nucleus resulted in retrogradely labeled neurons that were located in the postsubiculum, and also in the pre- and parasubiculum. These results are the first to establish the existence of a zinc-rich subicular-thalamic projection. Similar infusions in either the intralaminar nuclei or the zona incerta resulted in labeling of neurons in several brainstem structures related to the reticular formation. Our results provide morphological evidence for zinc modulation of glutamatergic inputs to highly selective thalamic nuclei, arising differentially from either cortical limbic areas or from brainstem ascending activation systems.

Fos expression in the rostral thalamic nuclei and associated cortical regions in response to different spatial memory tests

Using the quantification of the Fos protein as an indicator of neuronal activation, we studied the involvement of the rostral thalamic nuclei and associated structures in different spatial memory tasks in two experiments. In both experiments, tasks were matched for sensorimotor factors but differed in their spatial and mnemonic demands. In Experiment 1, matched groups of rats either ran in a standard eight-arm radial maze or ran up and down just one arm of the maze while the number of runs and rewards were matched across pairs of rats. In Experiment 2, both groups were trained on the eight-arm radial maze but in different rooms. On the test day, one group was moved so that both groups now performed the same radial-maze task in the same room but for one group the extramaze cues were novel. There were significant increases in Fos in all three of the anterior thalamic nuclei (anterodorsal, anteroventral and anteromedial) as well as the adjacent nucleus reuniens and rostral reticular thalamic nucleus, in both the eight-arm versus one-arm condition (Experiment 1) and the novel room versus familiar room condition (Experiment 2). There were no significant differences in the mediodorsal thalamic nucleus in either experiment. The more spatially demanding task in each experiment also resulted in increased Fos expression in the subicular complex (postsubiculum, presubiculum and parasubiculum), as well as in the prelimbic cortex. Performing the standard radial-arm maze task also produced significant Fos increases in both rostral and caudal levels of the retrosplenial cortex when compared to rats running up and down a single arm in the same maze (Experiment 1); performing the task in a novel room did not, however, result in any further Fos increases in this region (Experiment 2). The specificity of the changes in levels of Fos was shown by a lack of any consistent difference in levels in six control sites. The present results reveal a group of anatomically related structures that work together in the intact rat brain during tasks that tax allocentric spatial working memory.

Co‐Expression of Melatonin (MEL1a) Receptor and Arginine Vasopressin mRNAs in the Siberian Hamster Suprachiasmatic Nucleus

Durational melatonin signals, cued by the photoperiod and generated by the pineal gland, are processed in the brain to induce seasonally appropriate physiological and behavioural adaptations. The melatonin receptor subtype MEL1a (also known as mt1) appears to regulate seasonal responses. Single label in situ hybridization for MEL1a receptor mRNA revealed labelled cells in several brain regions of Siberian hamsters, including the suprachiasmatic nucleus, the paraventricular nucleus of the thalamus, and the reuniens nucleus of the thalamus. To characterize suprachiasmatic nucleus cells containing MEL1a receptor mRNA, we used 35S-labelled cRNA probes for MEL1a receptor mRNA in combination with digoxigenin-labelled cRNA probes for vasopressin, somatostatin, or orphan retinoid Z receptor beta (RZRbeta; a putative nuclear melatonin receptor). Cells in the suprachiasmatic nucleus that contained MEL1a receptor mRNA also contained mRNAs for vasopressin and RZRbeta, but not for somatostatin. These data suggest that suprachiasmatic nucleus vasopressin cells may respond to melatonin signals, raising the possibility that suprachiasmatic nucleus vasopressin output mediates some of the effects of melatonin on seasonal or circadian responses.

Distribution of luteinizing hormone-releasing hormone in the upper brainstem and diencephalon of the cat: An immunocytochemical study

The distribution of luteinizing hormone-releasing hormone (LH-RH)-immunostained cell bodies and fibres was studied in the brainstem and diencephalon of the cat using an indirect immunoperoxidase technique. The brainstem and the thalamus were devoid of immunostained cell bodies, whereas in the hypothalamus immunopositive perikarya were observed in the supraoptic nucleus, the anterior hypothalamus, the preoptic region and in the arcuate nucleus. Our findings also showed that the hypothalamus is richer in immunostained fibres, and that in this region such fibres are more widely distributed than in the thalamus and upper brainstem. No immunopositive fibres were observed in the lower brainstem. Our results point to a more widespread distribution of LH-RH-immunostained perikarya in the cat hypothalamus than that previously reported in the cat; a similar distribution to that found in the rat, and a more restricted distribution than in primates. Additionally, our study shows a more widespread distribution of immunostained fibres in the cat brainstem and diencephalon than that previously described for other mammals. In this context, our results describe for the first time in the mammals central nervous system fibres containing LH-RH located in the stria medullaris of the thalamus, the supramammillary decussation, the laterodorsal and lateroposterior thalamic nuclei, the nucleus reuniens, the supraoptic nucleus, and the optic chiasm. Thus, our findings reveal that LH-RH-immunostained structures are widely distributed in the upper brainstem and in the diencephalon of the cat, suggesting that the peptide may be involved in several physiological functions.

Differential distribution of metabotropic glutamate receptor subtype mRNAs in the thalamus of the rat

l-Glutamate ( l-Glu) is present in most excitatory synapses of the mammalian brain, acting on several receptor subtypes. Height different genes encoding metabotropic glutamate receptors (mGluRs) subtypes have been described (mGluR1–8), having a distinct distribution in the brain. In the present study, the distribution of mGluR1, 3, 4, 5 and 7 mRNAs was determined in 20 thalamic nuclei of adult rats by performing in situ hybridisation with subtype-specific 35 S -labelled oligonucleotide probes. High expression of mGluR1 mRNA mainly occurred in midline nuclei such as the centromedial/centrolateral (CM/CL) nuclei, parafascicular and submedius nuclei, and in the ventroposteromedial (VPM) and posterior (Po) nuclei. In contrast, mGluR5 mRNA was more uniformly distributed at weak to moderate levels, except in the reuniens nucleus where a strong signal was detected. The mGluR3 mRNA was highly expressed in the reticular thalamic nucleus and almost not detectable in any other thalamic region. Additionally, mGluR3 mRNA was found not only in neurones but also in putative glial cells. The mGluR4 mRNA was abundant in most thalamic nuclei, with prominent expression in the CM/CL, Po and ventrobasal complex (VPM and ventroposterolateral, VPL). Finally, mGluR7 transcripts were found evenly distributed throughout the thalamus at moderate levels, the highest signal being detected in the paraventricular thalamic nucleus, VPM, VPL and Po. This differential distribution of mGluR subtypes in the rat thalamus may contribute to the heterogeneity of glutamate effects on thalamic neurones. The mGluR1, mGluR4 and mGluR7 receptors may be involved in the processing of somatosensory information because they are expressed in nuclei that receive direct sensory input.

Nucleus reuniens thalami innervates γ aminobutyric acid positive cells in hippocampal field CA1 of the rat

The aim of the present study was to investigate whether the nucleus reuniens thalami (RE) innervates inhibitory cells in hippocampal field CA1. Therefore, we examined the RE-CA1 input at the ultrastructural level. RE axons were anterogradely labeled with biotin-conjugated dextran amine (BDA), in combination with pre-embedding γ aminobutyric acid (GABA)-immunolabeling of neurons in CA1. We observed that part of the BDA-labeled axons formed asymmetrical (i.e. excitatory) synapses on GABA-positive dendrites. Based on these data, which are in line with our previously published electrophysiological observations (Dolleman-Van der Weel, M.J., Lopes da Silva, F.H. and Witter, M.P., Nucleus reuniens thalami modulates activity in hippocampal field CA1 through excitatory and inhibitory mechanisms. J. Neurosci., 17 (1997) 5640–5650), we propose that RE-CA1 input partially influences hippocampal transmission through activation of local inhibitory interneurons.

Neurons Containing Hypocretin (Orexin) Project to Multiple Neuronal Systems

The novel neuropeptides called hypocretins (orexins) have recently been identified as being localized exclusively in cell bodies in a subregion of the tuberal part of the hypothalamus. The structure of the hypocretins, their accumulation in vesicles of axon terminals, and their excitatory effect on cultured hypothalamic neurons suggest that the hypocretins function in intercellular communication. To characterize these peptides further and to help understand what physiological functions they may serve, we undertook an immunohistochemical study to examine the distribution of preprohypocretin-immunoreactive neurons and fibers in the rat brain. Preprohypocretin-positive neurons were found in the perifornical nucleus and in the dorsal and lateral hypothalamic areas. These cells were distinct from those that express melanin-concentrating hormone. Although they represent a restricted group of cells, their projections were widely distributed in the brain. We observed labeled fibers throughout the hypothalamus. The densest extrahypothalamic projection was found in the locus coeruleus. Fibers were also seen in the septal nuclei, the bed nucleus of the stria terminalis, the paraventricular and reuniens nuclei of the thalamus, the zona incerta, the subthalamic nucleus, the central gray, the substantia nigra, the raphe nuclei, the parabrachial area, the medullary reticular formation, and the nucleus of the solitary tract. Less prominent projections were found in cortical regions, central and anterior amygdaloid nuclei, and the olfactory bulb. These results suggest that hypocretins are likely to have a role in physiological functions in addition to food intake such as regulation of blood pressure, the neuroendocrine system, body temperature, and the sleep-waking cycle.

Single midline thalamic neurons projecting to both the ventral striatum and the prefrontal cortex in the rat

The midline thalamic nuclei have been known to send projection fibres to the ventral striatum and the autonomic/limbic-associated areas of the prefrontal cortex. In the present study, we sought to determine whether or not single midline thalamic neurons project both to the ventral striatum and to the cerebral cortical areas. Experiments were performed on chloral hydrate-anaesthetized male Sprague–Dawley rats; two fluorescent retrograde tracers were centred on the medial or lateral part of the nucleus accumbens—the major part of the ventral striatum—and the medial or lateral prefrontal viscerolimbic cortex. Our retrograde double-labelling study revealed that a subset of midline thalamic neurons send projection fibres to both the nucleus accumbens and the cerebral cortex. Such neurons projecting to both targets were principally identified in the paraventricular thalamic nucleus. The majority of the dually-labelled neurons in the paraventricular thalamic nucleus projected to the lateral part of the nucleus accumbens and the medial wall of the prefrontal cortex. Dually-labelled neurons were additionally found in other midline nuclei, including the paratenial, intermediodorsal, rhomboid, and reuniens nuclei, as well as in the medial part of the parafascicular thalamic nucleus. Dually-projecting neurons identified in the present study may represent a potential link between the limbic striatum and the viscerolimbic-associated cortex, thus suggesting that non-discriminative information relayed to the prefrontal cortex might exert an influence through the same neurons on the nucleus accumbens implicated in affective behaviour.

Adrenergic innervation of the monkey thalamus: an immunohistochemical study

The distribution and function of the neurotransmitter adrenaline in the primate brain are poorly understood. Biochemical studies have shown the presence of adrenaline or its biosynthetic enzyme, phenylethanolamine- N-methyltransferase, in the rat and human thalamus. However, the distribution of the adrenergic fibres in the thalamus has only been demonstrated in rats. We study the adrenergic innervation of the macaque monkey thalamus using immunohistochemistry against phenyletanolamine- N-methyltransferase. The distribution of phenyletanolamine- N-methyltransferase-immunoreactive fibres is markedly heterogeneous and principally restricted to those nuclei, or their portions, that are located in or close to the midline, with the highest density being found in the paraventricular, parafascicular and mediodorsal nuclei. The paraventricular nucleus is densely innervated by adrenergic axons throughout, while the densest innervation of the parafascicular nucleus is located in its medial part and the strongest mediodorsal nuclear immunolabelling is found in its most posterior and medial region. Moderate or low concentrations of phenyletanolamine- N-methyltransferase-immunopositive fibres are present in the paratenial nucleus, and all parts of the central nucleus, nucleus reuniens, central medial nucleus, centromedian nucleus, medial geniculate body and medial pulvinar nucleus, while only scattered immunoreactive axons are found in other thalamic nuclei. The morphology of the phenyletanolamine- N-methyltransferase-immunoreactive axons is quite diverse, as they have different diameters and most are endowed with diversely-shaped varicosities. These findings are the first morphological evidence for the presence of adrenergic innervation in the primate thalamus and reveal that this innervation is highly selective, heterogeneous and more widely distributed in primates than in rats. The thalamic nuclei innervated by adrenaline are connected to widespread limbic and associative cortical areas as well as to subcortical structures, in particular the neostriatum and amygdala. We hypothesize that thalamic adrenaline may be implicated in emotional, social and attentional mechanisms through its facilitation of co-ordinated action by these brain regions.

Calretinin gene expression in the human thalamus

The localization and levels of expression of the calcium-binding protein calretinin (CR) in the human thalamus was studied with an in situ hybridization method applied to formalin-fixed postmortem material from normal individuals. The riboprobe used was generated from a specific fragment of human CR cDNA. As visualized on X-ray film autoradiographs, high levels of CR gene transcript occurred in several thalamic nuclei, including the reticular nucleus, mediodorsal nucleus, rostral intralaminar nuclei (paracentral, central medial and central lateral) and several midline nuclei (paraventricular, reuniens and medioventral nuclei). In the reticular nucleus, neurons expressing CR mRNA were few in number but formed dense and widely distributed clusters. In contrast, virtually all neurons in the rostral intralaminar and midline nuclei expressed very high levels of CR mRNA and formed a prominent rim around the mediodorsal nucleus, which contained scattered clusters of labeled neurons. The caudal intralaminar nuclei, principally the centromedian nucleus, displayed very few neurons expressing CR mRNA. Only the medial part of the parafascicular nucleus expressed moderate levels of CR mRNA. The nuclei of the ventral group (ventral anterior, lateral and posterior nuclei) were virtually devoid of CR gene transcript. This highly heterogeneous pattern of mRNA expression suggests that CR may be heavily involved in the function of the so-called non-specific nuclei, but not in that of the specific relay nuclei of the human thalamus. The data also demonstrate that the presence of CR gene transcript can easily be detected on formalin-fixed sections of the human brain.

Dorsocaudal SCN Microknife-Cuts Do Not Block Short Day Responses in Siberian Hamsters Given Melatonin Infusions

Siberian hamsters ( Phodopus sungorus sungorus ) undergo photoperiod-induced physiological and behavioral adaptations. These adaptations, including changes in reproductive and metabolic status, are triggered by the pineal gland through the nocturnal secretion of its principal hormone, melatonin. The possible CNS sites of melatonin action determined through radiolabeled melatonin binding include the paraventricular and reuniens nuclei of the thalamus and the suprachiasmatic nucleus (SCN). However, we do not know the mechanisms and circuitry involved in the transmission of melatonin signals. Bilateral electrolytic lesions of the SCN (SCNx) block the responses to short day–like (long duration) melatonin signals delivered daily via the timed infusion paradigm, suggesting that the SCN receives and transmits short-day melatonin signals. The purpose of the present experiment was to answer the following question: are short-day melatonin signals transmitted to other brain structures from the SCN through its dorsomedial/dorsocaudal fiber projections? Pinealectomized adult male hamsters given horizontal knife cuts (kc) just dorsocaudal to the SCN (SCN-kc), sham-kc, or SCNx were given daily subcutaneous short day–like melatonin infusions via the timed infusion paradigm for 6 weeks. Only the hamsters given SCNx exhibited long day–like gonadal, epididymal fat pad, and body masses. Therefore, short day melatonin signals received by the SCN were not transmitted to other areas of the central nervous system through SCN efferents projecting dorsomedially or dorsocaudally.

Reciprocal regional changes in brain NPY receptor density during dietary restriction and dietary-induced obesity in the rat

Neuropeptide Y (NPY) potently induces feeding, reduces thermogenesis and induces obesity in rats when injected into the cerebral ventricles. Groups of male Wistar rats were either restricted to 60% of their normal daily food intake over 10 days or made obese by presenting them with a high-calorie diet rich in sugars and fat over 6 weeks. Food restricted rats lost up to 20% of their body weight, compared with control rats and had large reductions in their body fat mass. By contrast, rats with dietary-induced obesity weighed 26% more than controls due mainly to increased body fat mass. Quantitative receptor autoradiography demonstrated reduced [ 125I]PYY binding in the hypothalamic lateral (perifornical) and dorsal areas, hypothalamic ventromedial, arcuate and dorsomedial nuclei, hippocampal CA3 region, centromedial amygdaloid nucleus and thalamic paraventricular and reuniens nuclei in dietary restricted rats compared with controls. By contrast, regional [ 125I]PYY binding was significantly increased in hypothalamic lateral and dorsal areas, hypothalamic arcuate and dorsomedial nuclei, amygdaloid medial and centromedial nuclei, thalamic centromedial and paraventricular nuclei of dietary obese rats versus controls. Masking NPY Y1 receptors with 1 μM BIBP3226, a selective Y1 receptor antagonist, revealed that the changes in [ 125I]PYY binding in brains of food-restricted and dietary-obese rats were due to changes in BIBP3226-insensitive binding sites, presumably Y2 or Y5 NPY receptors. These data suggest that dietary-restriction stimulates NPY release resulting in down-regulation of NPY Y5 `feeding' and/or Y2 receptors and reduced BAT thermogenesis thereby providing an increased drive to eat to restore normal caloric intake whilst reducing thermogenesis in order to conserve fat reserves. By contrast, the up-regulation of NPY Y5 and/or Y2 receptors in dietary-induced obesity is consistent with inhibition of NPY release in the hypothalamus, amygdala and thalamus. Overall, we suggest that there is a regional increase in NPY release during negative energy balance, such as during food-restriction and a reduced regional release of NPY in positive energy balance, such as during hyperphagia associated with the development of obesity.

Nucleus Reuniens Thalami Modulates Activity in Hippocampal Field CA1 through Excitatory and Inhibitory Mechanisms

The nucleus reuniens thalami (RE) originates dense projections to CA1, forming asymmetrical synapses on spines (50%) and dendrites (50%). The hypothesis that RE input modulates transmission in CA1 through excitation of both pyramidal cells and interneurons was tested using electrophysiological methods in the anesthetized rat. The RE-CA1 afferents were selectively stimulated at their origin; evoked field potentials and unit activity were recorded in CA1. RE-evoked depth profiles showed a prominent negative deflection in the stratum lacunosum-moleculare and a positive one in the stratum radiatum. The lacunosum-moleculare sink-radiatum source configuration is compatible with RE-elicited depolarization of apical dendrites of pyramidal cells. Despite a consistent and robust paired pulse facilitation of RE-evoked field potentials, population spikes in the stratum pyramidale were not detected at any tested condition. This indicates the inability of RE-CA1 input to discharge pyramidal cells. However, stimulation of RE-elicited spiking of extracellularly recorded units in strata oriens/alveus and distal radiatum, indicative of the activation of local interneurons. Thus, RE seems to modulate transmission in CA1 through a (subthreshold) depolarization of pyramidal cells and a suprathreshold excitation of putative inhibitory oriens/alveus and radiatum interneurons. RE-evoked monosynaptic or disynaptic field potentials were associated with stimulation of rostral or caudal RE, respectively. Anatomically, a projection from caudal to rostral RE was demonstrated that can account for the disynaptic RE-CA1 input. Because caudal RE receives input from the hippocampus via the subiculum, we propose the existence of a closed RE-hippocampal circuit that allows RE to modulate the activity in CA1, depending on hippocampal output.

Discrete thalamic lesions attenuate winter adaptations and increase body weight.

The midline thalamus (e.g., the paraventricular thalamic nuclei and the reuniens nucleus) of Siberian hamsters and other mammals has been reported to contain specific binding sites for melatonin, a hormone that is essential for photoperiodically induced winter adaptations such as reproductive quiescence, loss of body weight, daily torpor, and the winter molt. The first experiment investigated whether the midline thalamus is necessary for these winter adaptations. Adult Siberian hamsters received discrete neurotoxic lesions of the paraventricular thalamic nuclei or the reuniens nucleus while under pentobarbital sodium-induced anesthesia. After recovery, the hamsters were monitored for winter adaptations while they were exposed to short photoperiods (10 h light/day) for 12 wk at 22 degrees C then for 60 days at 7 degrees C. Lesions of the reuniens nucleus, but not of the paraventricular thalamic nuclei, significantly inhibited short photoperiod-induced loss of body weight and tended to increase food consumption and decrease daily torpor. The second experiment showed that lesions of the reuniens nucleus increased body weight gain compared with that in controls during exposure to long photoperiods at 22 degrees C for 16 wk. In summary, these findings show that the reuniens nucleus is an important site for regulation of body weight and suggest that lesions of the reuniens nucleus may attenuate winter metabolic adaptations by causing an increase in body weight.

Increased expression of the N-methyl- d-aspartate receptor subunit, NR1, in immunohistochemically identified magnocellular hypothalamic neurons during dehydration

N-Methyl- d-aspartate receptors are thought to be involved in synaptic signaling within the hypothalamo-neurohypophysial system, but the extent and nature of their involvement has not been determined. In this study, in the rat, we evaluated the effect of hyperosmotic stimulation on the N-methyl- d-aspartate receptor subunit, NR1, which confers function to N-methyl- d-aspartate receptor heteromers. Co-localization of immunoreactivity for NR1 and vasopressin- or oxytocin-associated neurophysin in magnocellular neurons of the supraoptic and paraventricular hypothalamic nuclei was accomplished using double-label immunohistochemistry. Our results show that vasopressin- and oxytocin–neurophysin-positive populations contained detectable levels of NR1 labeling. Using NR1 labeling as a measure of N-methyl- d-aspartate receptor density, we examined the effect of dehydration in these nuclei. Using computer-assisted densitometry, we found significantly greater NR1 labeling densities in the magnocellular regions of both the supraoptic and paraventricular nuclei of saline-treated rats than of control rats. This increase was not due to methodological factors, since no changes in NR1 labeling density were found in a nearby nucleus, the nucleus reuniens. Western blot analysis showed similar selective increases in NR1 labeling in homogenates from the supraoptic nucleus, paraventricular nucleus and in some cases from the anterior hypothalamic area. In both immunohistochemical and western blotting experiments we did not observe a dehydration-induced increase in NR1 in other brain areas examined. Our results showing an up-regulation of NR1-containing N-methyl- d-aspartate receptors during dehydration suggest that these receptors are involved in the regulation of body water and may represent an adaptive physiological response following activation of the hypothalamo-neurohypophysial axis. In addition, these results suggest that the functional expression of N-methyl- d-aspartate receptors is dynamic and may be modified according to the physiological state of the animal.