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Parasympathetic and sympathetic control of the pancreas: a role for the suprachiasmatic nucleus and other hypothalamic centers that are involved in the regulation of food intake.

To reveal brain regions and transmitter systems involved in control of pancreatic hormone secretion, specific vagal and sympathetic denervation were combined with injection of a retrograde transsynaptic tracer, pseudorabies virus (PRV), into the pancreas. After sympathetic or vagal transsection first-order neurons were revealed in the dorsal motor nucleus of the vagus (DMV) or in preganglionic spinal cord neurons (SPN), respectively. Careful timing of the survival of the animals allowed the detection of cell groups in immediate control of these DMV or SPN neurons. A far larger number of cell groups is involved in the control of DMV than of SPN neurons. Examples are given of a high level of interaction between the sympathetic and parasympathetic nervous system. Several cell groups project to both branches of the autonomic nervous system, sometimes even the same neurotransmitter is used, e.g., oxytocin neurons in the paraventricular nucleus and melanin-concentrating hormone and orexin neurons in the lateral hypothalamus project to both the DMV and SPN neurons. Moreover, the appearance of third-order neurons located in the sympathetic SPN after complete sympathectomy and in the DMV after complete vagotomy illustrates the possibility that motor neurons of the sympathetic and parasympathetic system may exchange information by means of interneurons. The presence of second-order neurons in prefrontal, gustatory, and piriform cortex may provide an anatomic basis for the involvement of these cortices in the cephalic insulin response. The observation that second-order neurons in both vagal and sympathetic control of the pancreas contain neuropeptides that are known to play a role in food intake indicates a direct association between behavioral and autonomic functions. Finally, the observation of third-order neurons in the suprachiasmatic nucleus and ventromedial hypothalamus shows the modulatory action of the time of the day and metabolic state, respectively.

Animals↗

Identification of an inward rectifier potassium channel gene expressed in mouse cortical astrocytes.

These experiments identify an inward rectifier K+ (Kir) channel expressed in mouse cortical and white matter astrocytes at the molecular level. Messenger RNA for one of the known Kir channel genes, Kir4.1, is present at much higher levels in cortical astrocytes in primary culture than the other known Kir family members. In culture, the level of Kir4.1 mRNA is lower in proliferating cells and in cells cultured for 16 h under hypoxic conditions, compared to confluent cells. Partial differentiation of the astrocytes with dibutyryl cAMP or by coculture with neurons has no effect on the Kir4.1 mRNA level. In situ hybridization experiments show that Kir4.1 mRNA is broadly distributed in the adult brain, including the neocortex, the stratum pyrimadale of the hippocampus, and the piriform cortex. Immunostaining confirms that the Kir4.1 protein is expressed by cultured astrocytes and also by cocultured cortical neurons. Astrocytes and neurons display a patchy pattern of immunostaining, raising the possibility that the channels sort themselves in clusters in the plasma membrane. Stellate cells in the neocortex and white matter are immunoreactive for Kir4.1, and double immunofluorescence experiments show colocalization of Kir4.1 and glial acidic fibrillary protein (GFAP) on stellate cells in the white matter. The cloned mouse Kir4.1 cDNA, when expressed heterologously in HEK cells, gives rise to inactivating Kir channels similar to those recorded from cultured astrocytes. These results indicate that the Kir4.1 gene product forms a Kir channel, or is a subunit of the channel, in mouse cortical astrocytes both in culture and in vivo.

Animals↗

Olfactory evolution and behavioral ecology in primates.

Primates are usually thought of as "visual" mammals, and several comparative studies have emphasized the role of vision in primate neural and sociocognitive specialization. Here I explore the role of olfactory systems, using phylogenetic analysis of comparative volumetric data. The relative sizes of the main olfactory bulb (MOB) and accessory olfactory bulb (AOB) tend to show different evolutionary patterns in accordance with their different functions. Although there is some evidence of correlated evolution of the two systems, this is apparent in only one clade (the strepsirhines). As predicted, the MOBs correlate predominantly with ecological factors (activity period and diet), while the AOBs correlate with social and mating systems. Related olfactory structures (i.e., the piriform cortex and amygdala) exhibit correlated evolution with the AOBs but not with the MOBs, and the corticobasolateral part of the amygdala exhibits a correlation with social group size in platyrrhines similar to that observed for the AOB. These social system correlations support the idea that there is an olfactory dimension to the concept of the social brain.

Animal Communication↗

Serial MRI after experimental febrile seizures: altered T2 signal without neuronal death.

Whereas most febrile seizures (FSs) carry a benign outcome, a subpopulation of individuals with prolonged FSs are at risk for later temporal lobe epilepsy. Signal changes on magnetic resonance imaging (MRI) may provide early markers for changes in neuronal integrity that may promote epileptogenesis in such individuals. Here, we used serial MRIs, obtained before and at several time points after experimental prolonged FSs, to determine the prevalence and distribution of signal changes on T2-weighted images and to investigate the pathological substrates leading to these changes. Seventy-five percent of immature rats with experimental prolonged FSs had abnormal T2 signal enhancement at 24 hours, and 87.5% at 8 days after the seizures. The altered T2 values involved the dorsal hippocampus (75%), the piriform cortex (87.5%), and the amygdala (25%). However, these changes were not accompanied by evidence of neuronal injury or death in these regions, as assessed using the Fluoro-Jade method. Thus, experimental prolonged FSs lead to relatively frequent abnormal MRI signal in "temporal lobe" structures. Although these changes do not signify cell death, they may denote pathological cellular processes that promote epileptogenesis. .

Animals↗

Thyrotropin-releasing hormone gene expression and receptors are differentially modified in limbic foci by seizures.

Previous studies using two seizure paradigms, electroconvulsive shock and kindling, suggested potential sites of endogenous thyrotropin-releasing hormone (TRH) action in specific epileptogenic areas. We studied TRH gene expression and TRH receptors in rat limbic areas using the kindling model of epilepsy. Immunoassayable TRH increased 4- to 20-fold over control levels in specific subregions of the hippocampus 24 hours after a single stage 5 seizure. Concurrently, TRH receptor binding was significantly reduced in hippocampal (23-39%) and amygdaloid (21-22%) membranes. Dramatic temporal and spatial changes in prepro-TRH messenger RNA were visualized by in situ hybridization histochemistry in the hippocampal dentate gyrus, the piriform cortex, and the amygdala. Peak hybridization occurred 6 and 12 hours postictally in these loci and returned toward basal levels by 24 hours. These results are consistent with the hypothesis that TRH may have an important role in the pathophysiology epilepsy by modulating excitatory processes.

Animals↗

Anatomical distribution of NPY-like immunoreactivity in the domestic chick brain (Gallus domesticus).

Neuropeptide Y-immunoreactive (NPY-ir) fibers and neurons in the brain of the domestic chick (Gallus domesticus) were described using an immunohistochemical technique. NPY-ir neurons were seen in the lobus parolfactorius; hyperstriatum, neostriatum, paleostriatum, and archistriatum; hippocampal and parahippocampal areas; dorsolateral corticoid area; piriform cortex; two thalamic areas contiguous to the n. rotundus; n. dorsolateralis anterior thalami, pars lateralis, and pars magnocellularis; n. periventricularis hypothalami; n. paraventricularis magnocellularis; regio lateralis hypothalami; n. infundibuli; inner zone of the median eminence; dorsal and lateral portions of the n. opticus basalis; n. raphes; and n. reticularis paramedianus. NPY-ir fibers were seen throughout the entire chick brain, but were more abundant in the hypothalamus where they formed networks and pathways. They were also observed in some circumventricular organs. The anatomical data of the present study regarding the distribution of NPY ir in the chick brain, together with the physiological findings of other studies, suggest that NPY plays a key role in the regulation of the neuroendocrine, vegetative, and sensory systems of birds by acting as a neuromodulator and/or neurotransmitter.

Animals↗

Central connections of the olfactory bulb in the American opossum (Didelphys virginiana): a light microscopic degeneration study.

Destructive lesions were made in the right olfactory bulb of 16 adult opossums. Following postoperative survival periods of 4 to 31 days, the animals were sacrificed and perfused with 10% Formalin. Frozen sections of the brain were cut in either the coronal, horizontal, or sagittal plane and processed by the Fink-Heimer II method. Degenerating axons of olfactory bulb neurons were traced caudally in the ipsilateral lateral olfactory tract (LOT). Small lesions revealed a topographic representation of the olfactory bulb within the LOT. The dorsal, lateral, and ventral parts of the bulb were, respectively, represented in the dorsal, intermediate, and ventral parts of the LOT. Terminal degeneration was observed in the superficial half of the molecular layer ipsilaterally in the following structures: anterior olfactory nucleus, anterior hippocampal rudiment, olfactory tubercle, piriform cortex, ventrolateral frontal neocortex, lateral entorhinal cortex, nucleus of the LOT, and the lateral aspect of the cortical amygdaloid nucleus. No degeneration was observed in the anterior limb of the anterior commissure. Dorsal and lateral parts of the olfactory bulb projected to the anterolateral aspect of the olfactory tubercle, whereas the ventral part projected heavily to the entire tubercle. There was no evidence of topographic projections to other olfactory structures. The observations of the present investigation indicated that the olfactory bulb projections in the opossum, a primitive mammal, are essentially comparable with those of placental mammals.

Animals↗

Expression pattern of alpha CaMKII in the mouse main olfactory bulb.

Multifunctional Ca(2+)/calmodulin-dependent protein kinase II (CaMKII) is highly enriched at synapses and has been implicated in regulating the formation and function of several sensory systems, including the visual and the somatosensory systems. Although there is evidence for CaMKII expression in the olfactory system, the cellular localization of CaMKII has not been well studied and its function remains unknown. In this study, we examined the normal expression patterns of the predominant alpha CaMKII in the mouse main olfactory bulb. We showed that alpha CaMKII expression levels were high in the olfactory bulb and were developmentally regulated. Immunoreactivity to alpha CaMKII was heavy in the external plexiform layer and the granule cell layer but minimal in the olfactory nerve layer and the glomerular layer. At the cellular level, alpha CaMKII was selectively expressed in the gamma-aminobutyric acid (GABA)ergic granule cells but not in the GABAergic periglomerular cells. Unexpectedly, alpha CaMKII was not detected in the glutamatergic mitral/tufted cells. At the ultrastructural level, alpha CaMKII immunoreactivity was positive in granule cell spines and dendrites, but negative in mitral/tufted cell dendrites. In contrast, in the piriform cortex, as in the majority of cortical regions, alpha CaMKII was expressed in the glutamatergic neurons but not in the GABAergic neurons. Our results set the stage for ongoing investigations of the roles of CaMKII in the formation and function of the olfactory system.

Animals↗

Immunohistochemical localization of candidates for vesicular glutamate transporters in the rat brain.

Vesicular glutamate transporter 1 (VGluT1) is one of the best markers for glutamatergic neurons, because it accumulates transmitter glutamate into synaptic vesicles. Differentiation-associated Na(+)-dependent inorganic phosphate cotransporter (DNPI) shows 82% amino acid identity to VGluT1, and is another candidate for vesicular glutamate transporters. Here, we report the immunocytochemical localization of DNPI and compare it with that of VGluT1 in the adult rat brain. Both DNPI and VGluT1 immunoreactivities were found mostly in neuropil, presumably in axon terminals, throughout the brain. In the telencephalic regions, intense DNPI immunoreactivity was observed in the glomeruli of the olfactory bulb, layer IV of the neocortex, granular layer of the dentate gyrus, presubiculum, and postsubiculum. In contrast, VGluT1 immunoreactivity was intense in the olfactory tubercle, layers I-III of the neocortex, piriform cortex, entorhinal cortex, hippocampus, dentate gyrus, and subiculum. In the thalamic nuclei, DNPI-immunoreactive terminal-like profiles were much larger than VGluT1-immunoreactive ones, suggesting that DNPI immunoreactivity was subcortical in origin. DNPI immunoreactivity was much more intense than VGluT1 immunoreactivity in many brainstem and spinal cord regions, except the pontine nuclei, interpeduncular nucleus, cochlear nuclei, and external cuneate nucleus. In the molecular layer of the cerebellar cortex, climbing-like fibers showed intense DNPI immunoreactivity, whereas neuropil contained dense VGluT1-immnoreactive deposits. Both DNPI and VGluT1 immunoreactivities were observed as mossy fiber terminal-like profiles in the cerebellar granular layer. DNPI and VGluT1 immunoreactivities appeared associated with synaptic vesicles in the axon terminals forming asymmetric synapses in several regions examined electron microscopically. The present results indicate that DNPI and VGluT1 are used by different neural components in most, if not all, brain regions, suggesting the complementary functions of DNPI and VGluT1.

Animals↗

Distribution of NTS3 receptor/sortilin mRNA and protein in the rat central nervous system.

The neurotensin (NT) receptor, NTS3, originally identified as the intracellular sorting protein sortilin, is a member of a recently discovered family of receptors characterized by a single transmembrane domain. The present study provides the first comprehensive description of the distribution of NTS3/sortilin mRNA and protein in adult rat brain using in situ hybridization and immunocytochemistry. Both NTS3/sortilin mRNA and immunoreactivity displayed a widespread distribution throughout the brain. High levels of NTS3/sortilin expression and immunoreactivity were found in neuronal cell bodies and dendrites of allocortical areas such as the piriform cortex and hippocampus. Regions expressing both high levels of NTS3/sortilin mRNA and protein also included several neocortical areas, the islands of Calleja, medial and lateral septal nuclei, amygdaloid nuclei, thalamic nuclei, the supraoptic nucleus, the substantia nigra, and the Purkinje cell layer of the cerebellar cortex. In the brainstem, all cranial nerve motor nuclei were strongly labeled. NTS3/sortilin mRNA and immunoreactivity were also detected over oligodendrocytes in major fiber tracts. Subcellularly, NTS3/sortilin was predominantly concentrated over intracytoplasmic membrane-bound organelles. Many of the areas exhibiting high levels of NTS3/sortilin (e.g., olfactory cortex, medial septum, and periaqueductal gray) have been documented to contain high concentrations of NT nerve cell bodies and axons, supporting the concept that NTS3/sortilin may play a role in NT sorting and/or signaling. Other areas (e.g., hippocampal CA fields, cerebellar cortex, and cranial nerve motor nuclei), however, are NT-negative, suggesting that NTS3/sortilin also exerts functions unrelated to NT signaling.

Adaptor Proteins, Vesicular Transport↗

Efferent connections of septal nuclei of the domestic chick (Gallus domesticus): an anterograde pathway tracing study with a bearing on functional circuits.

Small iontophoretic injections of the anterograde tracer Phaseolus vulgaris leucoagglutinin were placed in different subregions of the septum of domestic chicks. The main targets of septal projections comprised the ipsi- and contralateral septal nuclei, including the nucleus of the diagonal band, basal ganglia, including the ventral paleostriatum, lobus parolfactorius, nucleus accumbens, and olfactory tubercle, archistriatum, piriform cortex, and anterior neostriatum. Further diencephalic and mesencephalic septal projections were observed in the ipsilateral preoptic region, hypothalamus (the main regions of afferentation comprising the lateral hypothalamic nuclei, ventromedial, paraventricular and periventricular nuclei, and the mammillary region), dorsal thalamus, medial habenular and subhabenular nuclei, midbrain central gray, and ventral tegmental area. Contralateral projections were also encountered in the septal nuclei, ventral paleostriatum, periventricular and anteromedial hypothalamic nuclei, suprachiasmatic nucleus, and the lateral hypothalamic area. Avian septal efferents are largely similar to those of mammals, the main differences being a relatively modest hippocampal projection arising mainly from the nucleus of the diagonal band (as confirmed by a specific experiment with the retrograde pathway tracer True blue), the lack of interpeduncular projection, and a greater contingent of amygdalar efferents arising from the lateral septum rather than the nucleus of the diagonal band. This pattern of connectivity is likely to reflect an important role of the avian septal nuclei in the coordination of limbic circuits and the integration of a wide variety of information sources modulating the appropriate behavioral responses: attention and arousal level, memory formation, hormonally mediated behaviors, and their affective components (such as ingestive, reproductive, and parental behaviors), social interaction, locomotor modulation, and circadian rhythm.

Animals↗

Ontogeny of angiotensin II type 1 receptor mRNAs in fetal and neonatal rat brain.

Studies have demonstrated a specific function of the angiotensin II (Ang II) type 1 receptor (AT(1)) in regulation of adult central cardiovascular, fluid, and pituitary hormone release and a predominant role of the renin-angiotensin system in fetal and neonatal cardiovascular homeostasis. The pattern of brain AT(1) mRNA expression during fetal and neonatal development is currently unknown. We used radiolabeled cRNA probes for in situ hybridization histochemistry to determine the ontogenic development of the two AT(1) subtypes (AT(1a) and AT(1b)) mRNA in rat brain, from 11 days of gestation (E11) to 28 days after birth (P28). No AT(1b) mRNA was detected in the developing brain, whereas AT(1a) mRNA was first detected at E19. The age at which AT(1a) mRNA is first detected varied among different brain areas and expression predominates in areas involved in fluid homeostasis, pituitary hormone release, and cardiovascular regulation, where it persists until P28. AT(1a) mRNA expression is present from E19 onward in the median preoptic nucleus, the vascular organ of the lamina terminalis, the paraventricular nucleus, the periaqueductal gray, the nucleus raphe pallidus, the motor facial nucleus, and very weakly in the nucleus of the solitary tract and the ambiguous nucleus, and at E21 in the subfornical organ, the anterior olfactory nucleus and the piriform cortex. AT(1a) mRNA expression is present after birth in many regions, including the preoptic and lateral hypothalamic areas, the area postrema and medullary reticular nuclei. In conclusion, during brain development, expression of AT(1a) mRNA, appears in late gestation at E19, predominantly in forebrain areas involved in fluid homeostasis and cardiovascular regulation. In contrast, AT(1a) mRNA expression is absent or present only in very small amounts until after birth in many medullary nuclei, known to play an important role in cardiovascular modulation. Our results suggest that, in perinatal life, AT(1a) is involved in fluid and perhaps cardiovascular homeostasis and that the role of Ang II in modulating medullary cardiovascular centers matures later in postnatal life.

Aging↗

Efferent connections of the nucleus of the lateral olfactory tract in the rat.

The efferent connections of the nucleus of the lateral olfactory tract (LOT) were examined in the rat with the Phaseolus vulgaris leucoagglutinin (PHA-L) technique. Our observations reveal that layers II and III of LOT have largely segregated outputs. Layer II projects chiefly ipsilaterally to the olfactory bulb and anterior olfactory nucleus, bilaterally to the anterior piriform cortex, dwarf cell cap regions of the olfactory tubercle and lateral shell of the accumbens, and contralaterally to the lateral part of the interstitial nucleus of the posterior limb of the anterior commissure. Layer III sends strong bilateral projections to the rostral basolateral amygdaloid complex, which are topographically organized, and provides bilateral inputs to the core of the accumbens, caudate-putamen, and agranular insular cortex (dorsal and posterior divisions). Layer II projects also to itself and to layers I and II of the contralateral LOT, whereas layer III projects to itself, to ipsilateral layer II, and to contralateral layer III of LOT. In double retrograde labeling experiments using Fluorogold and cholera toxin subunit b tracers, LOT neurons from layers II and III were found to provide collateral projections to homonymous structures on both sides of the brain. Unlike other parts of the olfactory amygdala, LOT neither projects directly to the extended amygdala nor to the hypothalamus. Thus, LOT seemingly influences nonpheromonal olfactory-guided behaviors, especially feeding, by acting on the olfactory bulb and on ventral striatal and basolateral amygdaloid districts that are tightly linked to lateral prefrontal cortical operations.

Animals↗

Afferent and efferent connections of the dorsolateral corticoid area and a comparison with connections of the temporo-parieto-occipital area in the pigeon (Columba livia).

The dorsolateral corticoid area (CDL) in the pigeon telencephalon is a thin, superficial part of the caudal pallium adjoining the medially situated hippocampal formation. To determine the connectivity of CDL, and to distinguish CDL from the rostrally adjacent temporo-parieto-occipital area (TPO), injections of neural tracers were made into the caudal superficial pallium at various rostrocaudal levels. The results showed that injections caudal to A 6.75 (Karten and Hodos [1967] Baltimore: Johns Hopkins University Press) gave rise to reciprocal connections with subdivisions of the hippocampal formation, TPO, piriform cortex, posterior pallial amygdala, caudoventral nidopallium, densocellular part of the hyperpallium, lateral hyperpallium, frontolateral nidopallium, and lateral intermediate nidopallium. Of these, the hippocampal formation showed very strong connectivity with CDL, and projection fibers from CDL clearly separated the dorsomedial region of the hippocampal formation into lateral and medial portions. CDL projected directly to the olfactory bulb, but did not receive projections from it. In the diencephalon, CDL received efferents from a dorsal region of the medial part of the anterior dorsolateral nucleus of the thalamus, subrotundal nucleus, and internal paramedian nucleus of the thalamus. These findings suggest that CDL in the pigeon belongs to the limbic pallium and that in some respects it may be comparable to the mammalian cingulate cortex. In contrast, injections of tracers into the pallial surface at or rostral to A 7.00 showed marked differences in the pattern of both anterograde and retrograde labeling from that resulting from injections caudal to A 6.50, thereby indicating the approximate level of transition from CDL to TPO.

Afferent Pathways↗

Co-localization of phosphorylated extracellular signal-regulated protein kinases 1/2 (ERK1/2) and phosphorylated eukaryotic initiation factor 2alpha (eIF2alpha) in response to a threonine-devoid diet.

The anterior piriform cortex (APC) has been shown to be an essential brain structure for the detection of dietary indispensable amino acid (IAA) deficiency, but little has been known about possible molecular detection mechanisms. Increased phosphorylation of the alpha-subunit of the eukaryotic initiation factor 2alpha (eIF2alpha) has been directly linked to amino acid deficiency in yeast. Recently, we have shown increased phosphorylation of eIF2alpha (p-eIF2alpha) in the rat APC 20 minutes after ingestion of an IAA-deficient meal. We suggest that if phosphorylation of eIF2alpha is an important mechanism in detection of IAA deficiency, then APC neurons that show p-eIF2alpha should also show molecular evidence of potentiation. The present research demonstrates increased expression and co-localization of p-eIF2alpha and phosphorylated extracellular signal-regulated protein kinase 1/2 (p-ERK1/2) in APC neurons, but not in the primary motor or agranular insular cortices in response to an IAA-deficient diet. ERK1/2 is an element of the mitogen-activated protein kinase cascade, an intraneuronal signaling mechanism associated with neuronal activation. The region of the APC that responds to IAA deficiency with increased p-eIF2alpha and p-ERK1/2 labeling ranges from 3.1 to 2.5 mm rostral of bregma. Within this region, only a few neurons respond to IAA deficiency with co-localization of abundant p-eIF2alpha and p-ERK1/2. These chemosensory neurons probably detect IAA deficiency and generate neuronal signaling to other portions of the brain, changing feeding behavior.

Animal Feed↗

A study of the rat neuropeptide B/neuropeptide W system using in situ techniques.

In the rat, the neuropeptide B/neuropeptide W (NPB/NPW) system is composed of two ligands, neuropeptide B (NPB) and neuropeptide W (NPW), and one receptor, GPR7. Although preliminary analyses show roles in feeding, hormone secretion, and analgesia, the lack of a detailed anatomical map impairs our understanding of the NPB/NPW system. We demonstrate in this report the expression patterns of GPR7, NPB, and NPW precursor messenger ribonucleic acid (mRNA) in the rat brain by using in situ hybridization and in situ binding experiments. The amygdala expresses the highest levels of GPR7 mRNA and binding signals. Other nuclei with high levels of expression and binding are the suprachiasmatic and the ventral tuberomamillary nuclei. Moderate levels are seen in the dorsal endopiriform, dorsal tenia tecta, bed nucleus, and the red nucleus. Low levels are in the olfactory bulb, parastrial nucleus, hypothalamus, laterodorsal tegmentum, superior colliculus, locus coeruleus, and the nucleus of the solitary tract. Although the NPB precursor is mostly expressed at low levels in the brain, moderate expression is seen in anterior olfactory nucleus, piriform cortex, median preoptic nucleus, basolateral amygdala, hippocampus, medial tuberal nucleus, substantia nigra, dorsal raphe nucleus, Edinger-Westphal nucleus, and the locus coeruleus. To our surprise, the expression of NPW precursor was not detected. Our study greatly expands the preliminary in situ data previously reported. With this map of the NPB/NPW system in the rat brain, a better understanding of the functional implications of the system in various behavioral paradigms is now possible.

Animals↗

Differences in chemo- and cytoarchitectural features within pars principalis of the rat anterior olfactory nucleus suggest functional specialization.

The anterior olfactory nucleus (AON) lies between the olfactory bulb and piriform cortex and is the first bilaterally innervated structure in the olfactory system. It is typically divided into two subregions: pars externa and pars principalis. We examined the cytoarchitecture of pars principalis, the largest cellular area of the region, to determine whether it is homogeneously organized. Quantitative Nissl studies indicated that large cells (cell body area >2 standard deviations (SD) larger than the mean cell size) are densest in lateral and dorsolateral regions, while small cells (>1 SD smaller than the mean) are more numerous in medial and ventral areas. Further evidence for regional differences in the organization of the AON were obtained with immunohistochemistry for calbindin (CALB), parvalbumin (PARV), glutamic acid decarboxylase (GAD), and choline transporter (CHT). Cells immunopositive for CALB (CALB+) were denser in the deep portion of Layer II, although homogeneously dispersed throughout the circumference of the AON. PARV+ cells were located in the superficial half of Layer II and were sparse in ventral and medial regions. CHT+ and GAD+ fibers were denser in lateral versus medial regions. No regional differences were found in GAD+ somata, or in norepinephrine transporter or serotonin transporter immunoreactivity. The observed regional differences in cyto- and chemoarchitectural features may reflect functional heterogeneity within the AON.

Animals↗

Distribution of neuropeptide S receptor mRNA and neurochemical characteristics of neuropeptide S-expressing neurons in the rat brain.

Neuropeptide S (NPS) and its receptor (NPSR) constitute a novel neuropeptide system that is involved in regulating arousal and anxiety. The NPS precursor mRNA is highly expressed in a previously undescribed group of neurons located between the locus coeruleus (LC) and Barrington's nucleus. We report here that the majority of NPS-expressing neurons in the LC area and the principal sensory trigeminal nucleus are glutamatergic neurons, whereas many NPS-positive neurons in the lateral parabrachial nucleus coexpress corticotropin-releasing factor (CRF). In addition, we describe a comprehensive map of NPSR mRNA expression in the rat brain. High levels of expression are found in areas involved in olfactory processing, including the anterior olfactory nucleus, the endopiriform nucleus, and the piriform cortex. NPSR mRNA is expressed in several regions mediating anxiety responses, including the amygdaloid complex and the paraventricular hypothalamic nucleus. NPSR mRNA is also found in multiple key regions of sleep neurocircuitries, such as the thalamus, the hypothalamus, and the preoptic region. In addition, NPSR mRNA is strongly expressed in major output and input regions of hippocampus, including the parahippocampal regions, the lateral entorhinal cortex, and the retrosplenial agranular cortex. Multiple hypothalamic nuclei, including the dorsomedial and the ventromedial hypothalamic nucleus and the posterior arcuate nucleus, express high levels of NPSR mRNA, indicating that NPS may regulate energy homeostasis. These data suggest that the NPS system may play a key role in modulating a variety of physiological functions, especially arousal, anxiety, learning and memory, and energy balance.

Animals↗