Organization of adrenergic inputs to the paraventricular and supraoptic nuclei of the hypothalamus in the rat

Anterograde transport, retrograde transport, and immunohistochemical techniques were used to characterize the organization of neural inputs to the paraventricular (PVH) and supraoptic (SO) nuclei from the C1, C2, and C3 adrenergic cell groups in the rostral medulla. The results are as follows: 1) Phenylethanolamine-N-methyltransferase-immunoreactive (PNMT-IR) fibers and terminals were distributed to all parts of the parvicellular division of the PVH; the dorsal and dorsal medial subdivisions received the most prominent inputs, the lateral and ventral medial parts the least. Sparse terminal fields were found consistently in the magnocellular division of the PVH and in the SO. 2) A combined retrograde transport-immunohistochemical method was used to estimate the number and proportion of cells in the regions of the C1, C2, and C3 cell groups that contribute to the PNMT-IR innervation of the PVH. On average, 232 +/- 37 retrogradely labeled cells in the C1 cell group, 73 +/- 32 in the C2 cell group, and 96 +/- 26 in the C3 group stained positively for PNMT-IR. These values comprised 70%, 84%, and 89%, respectively, of all retrogradely labeled neurons in these regions. 3) Fibers and terminals arising from the regions of each of the three adrenergic cell groups were labeled by local injections of the anterogradely transported plant lectin PHA-L. Each component projection was found to distribute in a very similar fashion and to mimic the overall distribution of PNMT-IR; differential projection patterns within the PVH or SO were not seen consistently following deposits in any of the individual adrenergic cell groups or at different rostrocaudal levels of any individual cell group. 4) A dual anterograde tracing (PHA-L)-immunohistochemical (PNMT) labeling method revealed an appreciable number of varicosities arising from the regions of C1, C2, and C3 cell groups to contain PNMT-IR. These results suggest that adrenergic inputs to the PVH and SO, while arising from distinct medullary cell groups and presumably relaying different types of sensory information, are in a position to influence similar groups of parvicellular neurosecretory and/or autonomic-related projection neurons.     

Anatomical specificity of noradrenergic inputs to the paraventricular and supraoptic nuclei of the rat hypothalamus

The distribution of neural inputs to the paraventricular (PVH) and supraoptic (SO) nuclei from the regions of the A1, the A2, and the A6 (locus coeruleus) noradrenergic cell groups was investigated by using a plant lectin, Phaseolus vulgaris leucoagglutinin (PHA-L), as an anterogradely transported tracer. An immunofluorescence double-labeling procedure was used to determine the extent to which individual anterogradely labeled fibers and terminals in the PVH and the SO also displayed immunoreactive dopamine-beta-hydroxylase (DBH), a marker for catecholaminergic neurons. The results may be summarized as follows: (1) Projections from the A1 region were found primarily, and in some experiments almost exclusively, in those parts of the magnocellular division of the PVH and the SO known to contain vasopressinergic neurons. (2) Projections from the A2 region were distributed primarily throughout the parvicellular division of the PVH and were most dense in the dorsal medial part, a region known to contain a prominent population of corticotropin-releasing factor (CRF)-immunoreactive neurons. In addition, a less-dense projection to the magnocellular division of the PVH and to the SO was consistently found. (3) Fibers originating from the locus coeruleus were distributed almost exclusively to the parvicellular division of the PVH, with the most prominent input localized to the periventricular zone, a part of the PVH known to contain dopamine-, somatostatin-, and thyrotropin-releasing-hormone-containing neurons. We found no evidence for a projection from A6 to the SO. (4) The majority of fibers originating from the A1, the A2 or the A6 regions contained DBH immunoreactivity, although an appreciable number did not. These results suggest that each of the three brainstem noradrenergic cell groups that contribute to the innervation of the PVH and/or the SO is in a position to modulate the activity of anatomically and chemically distinct groups of neurosecretory neurons.     

Prolactin and oxytocin interaction in the paraventricular and supraoptic nuclei: effects on oxytocin mRNA and nitric oxide synthase

We investigated the contribution of prolactin and oxytocin to the increase in staining for NADPH-d and oxytocin mRNA in the paraventricular nucleus (PVN) and supraoptic nucleus (SON) observed at the end of pregnancy, or following a steroid-priming regimen that mimics the hormonal profile of late pregnant females. Ovariectomized rats received chronic implants of silastic capsules containing oestrogen and progesterone followed by progesterone removal. In experiment 1, oxytocin antagonist (OTA) was administered to rats to investigate whether intranuclear oxytocin release was necessary for NADPH-d staining. In experiments 2a and b, rats received concurrent treatment with bromocryptine (0.5 mg/day) to suppress endogenous prolactin release, and either systemic prolactin (0.5 mg once daily), or prolactin (2 micro g/ micro l), or vehicle infused twice a day into the third ventricle, or chronic oxytocin infusion (24 ng/day) for 3 days following progesterone removal. Brains were then processed for NADPH-d histochemistry. In experiment 3, the interaction of prolactin and oxytocin on oxytocin mRNA within the SON and PVN was examined. NADPH-d staining in the SON and PVN was reduced by the highest dose of the OTA, and by bromocryptine treatment. Central prolactin and oxytocin replacement completely restored NADPH-d staining in bromocryptine-treated rats. Finally, both bromocryptine and the OTA suppressed oxytocin mRNA expression and prolactin replacement restored expression levels to that of controls. Together, these data suggest that the increased capacity to produce nitric oxide in the SON and PVN during late pregnancy is dependent on prolactin stimulating oxytocin gene mRNA and hence intranuclear oxytocin release.     

Localization of kappa opioid receptors in oxytocin magnocellular neurons in the paraventricular and supraoptic nuclei

It has been demonstrated previously that kappa opioid receptor agonists, such as dynorphin, inhibit oxytocin secretion in the rat. To determine whether kappa agonists act directly on oxytocin-containing magnocellular neurons to inhibit hormone secretion, we utilized immunofluorescence to examine the cellular localization of kappa opioid receptors in the rat paraventricular and supraoptic nuclei. kappa Opioid receptor immunoreactivity co-localized with oxytocin-containing cell bodies, their axons and axon terminals. Thus, our results suggest that kappa opioid receptor agonists can exert direct inhibitory actions on oxytocin magnocellular neurons.     

Effects of lesions in the hypothalamic paraventricular, supraoptic and suprachiasmatic nuclei on vasopressin and oxytocin in rat brain and spinal cord

The content of arginine vasopressin and oxytocin in various extrahypothalamic sites of the rat brain and spinal cord was determined by specific radioimmunoassays after lesions had been made in either the paraventricular (PVN), supraoptic (SON) or suprachiasmatic nuclei (SCN). In some animals all 3 nuclei were destroyed together. The PVN provided a considerable amount of the vasopressin innervation of the solitary tract nucleus, and most of that in the spinal cord. Oxytocin was removed from some areas after lesions of the PVN and, again, most of this peptide was lost from the spinal cord. Lesions of the SCN did not appear to be followed by significant quantitative changes in either hormone in any of the areas studied. Lesions of the SON resulted in loss of oxytocin, particularly in the periventricular grey and some other areas, suggesting that extrahypothalamic projections from this nucleus may be more important than was previously assumed. Lesions of all 3 nuclei which included destruction of accessory hypothalamic nuclei resulted in a much more widespread loss of vasopressin and oxytocin, but there was preservation of both peptides in the dorsal raphe nucleus and much of those present in the locus coeruleus. It is concluded that the contribution of the classical hypothalamic nuclei to the extrahypothalamic content of vasopressin and oxytocin in rat brain is less than was originally believed, and that there are areas of the brain such as the locus coeruleus and dorsal raphe nucleus in which the source of these peptides may be outside the hypothalamus.     

Localization of κ opioid receptors in oxytocin magnocellular neurons in the paraventricular and supraoptic nuclei

It has been demonstrated previously that κ opioid receptor agonists, such as dynorphin, inhibit oxytocin secretion in the rat. To determine whether κ agonists act directly on oxytocin-containing magnocellular neurons to inhibit hormone secretion, we utilized immunofluorescence to examine the cellular localization of κ opioid receptors in the rat paraventricular and supraoptic nuclei. κ Opioid receptor immunoreactivity co-localized with oxytocin-containing cell bodies, their axons and axon terminals. Thus, our results suggest that κ opioid receptor agonists can exert direct inhibitory actions on oxytocin magnocellular neurons.     

The neurotensinergic synaptic innervation of vasopressin containing neurons in the rat hypothalamic paraventricular nucleus

A recent physiological report suggested that neurotensin could inhibit the vasopressin releasing from vasopressin-producing neurons in the hypothalamic paraventricular nucleus but not in the supraoptic nucleus. In the present study, the synaptic relationship between the neurotensin-like immunoreactive and vasopressin-like immunoreactive neurons has been examined using a pre-embedding double immunostaining technique in the rat hypothalamic paraventricular nucleus. At the light microscopic level, many neurotensin-like immunoreactive fibers were found near the vasopressin-like immunoreactive neurons. At the electron microscopic level, the neurotensin-like immunoreactive fibers were identified as axon terminals that made many synapses on the vasopressin-like immunoreactive perikarya and dendrites. The synapses were both asymmetrical and symmetrical. These findings of the present study suggest that the inhibitory effect of neurotensin on the vasopressin neurons in the hypothalamic paraventricular nucleus may be due to the direct synapses made by neurotensin-like immunoreactive axon terminals on the vasopressin-like immunoreactive neurons.     

Magnocellular neurosecretory neurons with ferritin-like immunoreactivity in the hypothalamic supraoptic and paraventricular nuclei of the rat.

Immunohistochemistry for rat liver ferritin (FRT) revealed an intensive labeling in some structures of the rat brain. In the supraoptic (SON) and paraventricular (PVN) hypothalamic nuclei, almost all neurosecretory neurons with vasopressin (AVP)-like immunoreactivity were immunostained with FRT. After water deprivation, a marked enlargement of cell body and an immunoreactivity to transferrin receptors were found in AVP-, FRT- and double (AVP+FRT)-labeled neurons in the SON and PVN.     

Ultrastructural localization of inhibin β- and somatostatin-28-immunoreactivities in the paraventricular and supraoptic nuclei

Recent studies have supported the existence of projections to the paraventricular and supraoptic nuclei of the hypothalamus that arise from non-catecholaminergic neurons in the nucleus of the solitary tract, whose terminal distribution is suggestive of interactions with both parvocellular and magnocellular neurosecretory neurons. Pre-embedding immunolabeling methods were used to compare and characterize the termination patterns of axons immunoreactive for two putative markers for this projection system, inhibin β and somatostatin-28, at the ultrastructural level. Axon terminal profiles stained fro either peptide were found to form symmetric or asymmetric junctions predominantly with the shafts of unlabeled dendrites of varying caliber. A small percentage of peptidergic terminals was found in both hypothalamic nuclei to engage in so-called ‘shared synapses’, where a single terminal profile contacted two postsynaptic elements. Axo-somatic terminations were relatively rarely seen in the supraoptic nucleus, but were somewhat more abundant in the paraventricular nucleus. These comprised principally symmetric junctions onto the somatic membranes of an ostensibly mixed population of cells, some of which bore apparent neurosecretory specializations. Combined immunoperoxidase and immuno-autoradiographic staining methods were used to estimate the extent to which either terminal type interacts with oxytocin neurons. Oxytocin stained elements comprised a minority of the postsynaptic targets of both peptidergic terminal types in the paraventricular nucleus, and a scant majority of those in the supraoptic nucleus. These results support the view that peptidergic neurons in the caudal nucleus of the solitary tract interact synaptically with multiple cell types in the parvocellular division of the paraventricular nucleus, and preferentially with oxytocinergic elements in the magnocellular neurosecretory system.     

Central administration of neuropeptide FF causes activation of oxytocin paraventricular hypothalamic neurones that project to the brainstem

Neuropeptide FF (NPFF), a morphine modulatory peptide, is emerging as an important neuromodulator in the context of central autonomic and neuroendocrine regulation. NPFF immunoreactivity and receptors have been identified in discrete autonomic regions within the brain and spinal cord, including the hypothalamic paraventricular nucleus (PVN). In this study, we examined the effects of intracerebroventricular (i.c.v.) administration of NPFF on activation of chemically identified PVN neurones that project to the brainstem nucleus of the solitary tract (NTS). In conscious rats, i.c.v. NPFF at a dose of 10 micro g, but not 8 micro g, caused an increase in arterial blood pressure. Immunohistochemical analysis revealed a dose-dependent increase in activated (Fos positive) PVN neurones following i.c.v. NPFF administration compared to controls receiving i.c.v. saline. Activated PVN neurones were located predominantly in the parvocellular compartment of the nucleus with relatively few Fos positive cells in the magnocellular subdivision. Chemical identification of activated neurones revealed significant number of activated cells to be oxytocin positive, whereas only few vasopressin, tyrosine hydroxylase (TH) or corticotrophin-releasing factor (CRF) neurones were double-labelled. Injection of the retrograde tracer fluorogold into the NTS resulted in labelling of significant numbers of parvocellular oxytocin, but not vasopressin, TH or CRF, PVN neurones. We conclude that centrally administered NPFF stimulates brainstem-projecting oxytocin PVN neurones. Oxytocin released from terminals within the NTS oxytocin thus modulate the activity of ascending visceral autonomic pathways that synapse initially within the NTS.     

Glutamate agonists activate the hypothalamic-pituitary-adrenal axis through hypothalamic paraventricular nucleus but not through vasopressinerg neurons

The hypothalamic-pituitary-adrenal (HPA) axis plays a crucial role in the stress processes. The nucleus paraventricularis hypothalami (PVN) with corticotropin-releasing hormone (CRH)-containing and arginine vasopressin (AVP)-containing neurons is the main hypothalamic component of the HPA. The glutamate, a well-known excitatory neurotransmitter, can activate the HPA inducing adrenocorticotropin hormone (ACTH) elevation. The aim of our study was to examine the involvement of PVN and especially AVP in glutamate-induced HPA activation using agonists of the N-methyl-d-aspartate (NMDA) and kainate receptors. Two approaches were used: in male Wistar rats the PVN was lesioned, and AVP-deficient homozygous Brattleboro rats were also studied. Blood samples were taken through indwelling cannula and ACTH, and corticosterone (CS) levels were measured by radioimmunoassay. The i.v. administered NMDA (5 mg/kg) or kainate (2.5 mg/kg) elevated the ACTH and CS levels already at 5 min in control (sham-operated Wistar or heterozygous Brattleboro) rats. The PVN lesion had no influence on basal ACTH and CS secretion but blocked the NMDA- or kainate-induced ACTH and CS elevations. The lack of AVP in the Brattleboro animals had no significant influence on the basal or glutamate-agonists-induced ACTH and CS elevations. Our results suggest that NMDA and kainate may activate the HPA axis at central (PVN) level and not at the level of pituitary or adrenal gland and that AVP has minor role in glutamate-HPA axis interaction. The time course of the ACTH secretion was different with NMDA or kainate. If we compared the two curves, the results were not coherent with the general view that NMDA activation requires previous kainate activation. Although it has to be mentioned that the conclusion which can be drawn is limited, the bioavailability of the compounds could be different as well.     

Forced swimming stimulates the expression of vasopressin and oxytocin in magnocellular neurons of the rat hypothalamic paraventricular nucleus.

Previous studies have shown that a 10-min forced swimming session triggers the release of both vasopressin and oxytocin into the extracellular fluid of the hypothalamic paraventricular (PVN) and supraoptic nuclei (SON) in rats. At the same time oxytocin, but not vasopressin, was released from the axon terminals into the blood. Here we combined forced swimming with in situ hybridization to investigate whether (i) the stressor-induced release of vasopressin and oxytocin within the PVN originates from parvo- or magnocellular neurons of the nucleus, and (ii) central release with or without concomitant peripheral secretion is followed by changes in the synthesis of vasopressin and/or oxytocin. Adult male Wistar rats were killed 2, 4 or 8 h after a 10-min forced swimming session and their brains processed for in situ hybridization using 35S-labelled oligonucleotide probes. As measured on photo-emulsion-coated slides, cellular vasopressin mRNA concentration increased in magnocellular PVN neurons 2 and 4 h after swimming (P < 0.05). Similarly, oxytocin mRNA concentration was significantly increased in magnocellular neurons of the PVN at 2 and 8 h (P < 0.05). We failed to observe significant effects on vasopressin and oxytocin mRNA levels in the parvocellular PVN and in the SON. Taken together with results from previous studies, our data suggest that magnocellular neurons are the predominant source of vasopressin and oxytocin released within PVN in response to forced swimming. Furthermore, in the case of vasopressin, central release in the absence of peripheral secretion is followed by increased mRNA levels, implying a refill of depleted somato-dendritic vasopressin stores. Within the SON, however, mRNA levels are poor indicators of the secretory activity of magnocellular neurons during stress.     

Branching projections of catecholaminergic brainstem neurons to the paraventricular hypothalamic nucleus and the central nucleus of the amygdala in the rat

In this study, we have employed triple fluorescent-labelling to reveal the distribution of catecholaminergic neurons within three brainstem areas which supply branching collateral input to the central nucleus of the amygdala (CNA) and the hypothalamic paraventricular nucleus (PVN): the ventrolateral medulla (VLM), the nucleus of the solitary tract (NTS) and the locus coeruleus (LC). The catecholaminergic identity of the neurons was revealed by immunocytochemical detection of the biosynthetic enzyme, tyrosine hydroxylase. The projections were defined by injections of two retrograde tracers, rhodamine- and fluorescein-labelled latex microspheres, in the CNA and PVN, respectively. In the VLM and NTS, the greatest incidence of neurons which contained both retrograde tracers was found at the level of the area postrema. These neurons were mainly located within the confines of the A1/C1 (VLM) and A2 (NTS) catecholaminergic neuronal groups. Double-projecting neurons in the LC (A6) were distributed randomly within the nucleus. It was found that 15% in the VLM, 10% in the NTS and 5% in the LC of the retrogradely labelled cells projected via branching collaterals to the PVN and CNA. One half of these neurons in the VLM and NTS were catecholaminergic, in contrast to the LC where virtually all double-retrogradely labelled neurons revealed tyrosine hydroxylase immunoreactivity. In the other brainstem catecholaminergic cell groups (A5, A7, C3), no catecholaminergic neurons were found that supplied branching collaterals to the CNA and PVN. Our results indicate that brainstem neurons may be involved in the simultaneous transmission of autonomic-related signals to the CNA and the PVN. Catecholamines are involved in these pathways as chemical messengers. Brainstem catecholaminergic and non-catecholaminergic neurons, through collateral branching inputs may provide coordinated signalling of visceral input to rostral forebrain sites. This may lead to a synchronized response of the CNA and PVN for the maintenance of homeostasis.     

Colocalization of calretinin and calbindin-D28k with oxytocin and vasopressin in rat supraoptic nucleus neurons: A quantitative study

Recent electrophysiological experiments, in which purified calbindin-D_28k (calbindin) and calretinin antibodies were diffused into these neurons, showed that Ca²⁺-dependent membrane potentials and firing patterns were profoundly and predictably affected by Ca²⁺-binding proteins (CaBPs). The present study used quantitative analyses of a dual-labeling immunofluorescence method to investigate the colocalization of the CaBPs, calbindin and calretinin in oxytocin (OT)- and (VP)-containing neurons of the supraoptic nucleus. Analyses of tissue immunostained with two different dilutions of each CaBP antibody used, revealed that 84% and 72% of the OT neurons were positive for calbindin immunoreactivity (-ir) at the higher and lower antibody concentrations, respectively. 52% and 50% of OT neurons were positive for calretinin-ir; thus, many OT neurons express both calbindin and calretinin. In contrast, only 25% and 18% of VP neurons showed calbindin-ir, and they were virtually devoid of calretinin-ir. These results provide evidence that CaBP expression in OT neurons is both greater and more diverse than in VP neurons, and are consistent with the hypothesis that Ca²⁺ buffering capacity contributes to the control of intrinsic firing patterns.     

The hypothalamic supraoptic and paraventricular nuclei of the echidna and platypus

The monotremes are an intriguing group of mammals that have major differences in their reproductive physiology and lactation from therian mammals. Monotreme young hatch from leathery skinned eggs and are nourished by milk secreted onto areolae rather than through nipples. Parturition and lactation are in part controlled through the paraventricular and supraoptic nuclei of the hypothalamus. We have used Nissl staining, enzyme histochemistry, immunohistochemistry for tyrosine hydroxylase, calbindin, oxytocin, neurophysin and non-phosphorylated neurofilament protein, and carbocyanine dye tracing techniques to examine the supraoptic and paraventricular nuclei and the course of the hypothalamo-neurohypophysial tract in two monotremes: the short-beaked echidna (Tachyglossus aculeatus) and the platypus (Ornithorhynchus anatinus). In both monotremes, the supraoptic nucleus consisted of loosely packed neurons, mainly in the retrochiasmatic position. In the echidna, the paraventricular nucleus was quite small, but had similar chemoarchitectural features to therians. In the platypus, the paraventricular nucleus was larger and appeared to be part of a stream of magnocellular neurons extending from the paraventricular nucleus to the retrochiasmatic supraoptic nucleus. Immunohistochemistry for non-phosphorylated neurofilament protein and carbocyanine dye tracing suggested that hypothalamo-neurohypophysial tract neurons in the echidna lie mainly in the retrochiasmatic supraoptic and lateral hypothalamic regions, but most neurophysin and oxytocin immunoreactive neurons in the echidna were found in the paraventricular, lateral hypothalamus and supraoptic nuclei and most oxytocinergic neurons in the platypus were distributed in a band from the paraventricular nucleus to the retrochiasmatic supraoptic nucleus. The small size of the supraoptic nucleus in the two monotremes might reflect functional aspects of monotreme lactation.     

Hypovolemia induces Fos-like immunoreactivity in neurons of the rat supraoptic and paraventricular nuclei.

Immunoreactivities to Fos proteins were detected in numerous neurons in the supraoptic, paraventricular and accessory neurosecretory nuclei 1 h following withdrawal of 4-5 cc of blood from the rat femoral arteries. Few or no positive cells were observed in the same nuclei in sham-operated or control animals. It is concluded that hypovolemia induces c-fos expression in hypothalamic neurons known to be associated with blood volume/pressure regulation.     

Convergent influence of the central nucleus of the amygdala and the paraventricular hypothalamic nucleus upon brainstem autonomic neurons as revealed by c-fos expression and anatomical tracing

Combinations of anatomical tracing with detection of Fos (the protein product of the immediate early gene c-fos) consequent to the stimulation of the central nucleus of the amygdala were used to explore the possibility that the hypothalamic paraventricular nucleus participates in the activation of brainstem neurons in the nucleus of the solitary tract and ventrolateral medulla. After injections of the anterograde tracer Phaseolus vulgaris leucoagglutinin in the paraventricular nucleus, labeled fibers and varicosities were found to impinge on catecholaminergic and non-catecholaminergic Fos-positive neurons in the brainstem. After injections of a retrograde tracer in the nucleus of the solitary tract or ventrolateral medulla, we observed that some of the Fos-positive neurons within the parvocellular paraventricular nucleus that project to the brainstem were catecholaminergic or oxytocinergic. The results indicate that direct and indirect inputs from the amygdala may influence the activity of autonomic neurons in the brainstem. The paraventricular nucleus, via its direct projections onto catecholaminergic and non-catecholaminergic neurons, may participate in activation of brainstem neurons. Activated catecholaminergic and oxytocinergic parvocellular neurons in the paraventricular nucleus may be involved in the transmission of autonomic signals from the amygdala toward the brainstem. ©1995 Wiley-Liss, Inc.     

Cyto- and chemoarchitecture of the hypothalamic paraventricular nucleus in the C57BL/6J male mouse: a study of immunostaining and multiple fluorescent tract tracing

The paraventricular nucleus of the hypothalamus (PVH) plays a critical role in the regulation of autonomic, neuroendocrine, and behavioral activities. This understanding has come from extensive characterization of the PVH in rats, and for this mammalian species we now have a robust model of basic PVH neuroanatomy and function. However, in mice, whose use as a model research animal has burgeoned with the increasing sophistication of tools for genetic manipulation, a comparable level of PVH characterization has not been achieved. To address this, we employed a variety of fluorescent tract tracing and immunostaining techniques in several different combinations to determine the neuronal connections and cyto- and chemoarchitecture of the PVH in the commonly used C57BL/6J male mouse. Our findings reveal a distinct organization in the mouse PVH that is substantially different from the PVH of male rats. The differences are particularly evident with respect to the spatial relations of two principal neuroendocrine divisions (magnocellular and parvicellular) and three descending preautonomic populations in the PVH. We discuss these data in relation to what is known about PVH function and provide the work as a resource for further studies of the neuronal architecture and function of the mouse PVH.     

Activation of neurons projecting to the paraventricular hypothalamic nucleus by intravenous lipopolysaccharide

The central nervous system interacts with the immune system to coordinate several components of the acute phase response, although the specific neuroanatomical pathways that mediate these responses are still uncharacterized. However, neurons in both the autonomic and endocrine components of the paraventricular hypothalamic nucleus (PVH) are characteristically activated in different models of immune stimulation. In the current study, we have used intravenous administration of lipopolysaccharide (LPS; 5 or 125 μg/kg) to induce the acute phase response. We subsequently coupled immunohistochemistry for Fos (as a marker of neuronal activation) with retrograde transport of the neuroanatomical tracer cholera toxin-b from the PVH. Several of the activated cell groups directly projected to the paraventricular nucleus, including the visceromotor (infralimbic) cortex, median preoptic nucleus, ventrome-dial preoptic area, bed nucleus of the stria terminalis, parabrachial nucleus, ventrolateral medulla, and nucleus of the solitary tract. These findings indicate that immune system stimulation activates cell groups from multiple nervous system levels that project to the paraventricular nucleus. We hypothesize that the activation of specific autonomic and endocrine elements of the PVH may be due to the activity of distinct afferents that converge on the PVH from multiple components of the central autonomic control system. Our results are consistent with the hypothesis that the PVH plays a key role in integrating diverse physiological cues into the varied manifestations that constitute the cerebral component of the acute phase response. © 1996 Wiley-Liss, Inc.