Larry W. Swanson

Organization of Ovine Corticotropin-Releasing Factor Immunoreactive Cells and Fibers in the Rat Brain: An Immunohistochemical Study

The distribution of corticotropin-releasing factor (CRF)-immunoreactive cells and fibers has been examined in the brains of normal adult rats, and in the brains of animals that had been pretreated with intraventricular injections of colchicine, or had been adrenalectomized 3-60 days before perfusion. The results suggest that CRF immunoreactivity is localized in at least three functionally distinct systems. First, most of the CRF-stained fibers in the neurohemal zone of the median eminence, which presumably modulate the release of ACTH and beta-endorphin from the pituitary, appear to arise in the paraventricular nucleus of the hypothalamus (PVH). About 2,000 CRF-stained cells are distributed throughout all eight parts of the PVH, although a majority (80%) of the cells are concentrated in the parvocellular division, and a smaller number (about 15%) are found in parts of the magnocellular division in which oxytocinergic cells predominate. This appears to be the only CRF-stained pathway in the brain that is affected (increased staining intensity) by adrenalectomy. Second, a series of cell groups in the basal telencephalon, hypothalamus, and brain stem that are known to play a role in the mediation of autonomic responses contain CRF-stained neurons. These areas, which are interconnected by stained fibers in the medial forebrain bundle and the periventricular system, include the central nucleus of the amygdala, substantia innominata, bed nucleus of the stria terminalis, medial and lateral preoptic areas, lateral hypothalamic area, central gray, laterodorsal tegmental nucleus, locus ceruleus, parabrachial nucleus, dorsal vagal complex, and regions containing the A1 and A5 catecholamine cell groups. And third, scattered CRF-stained cells are found throughout most areas of the cerebral cortex. Most such cells are confined to layers II and III in the neocortex, and their bipolar shape suggests that they are interneurons. These cells are most common in limbic regions including prefrontal areas, the cingulate gyrus, and areas bordering the rhinal fissure. Scattered immunoreactive cells are also found in dorsal parts of the dentate gyrus and Ammons horn. These results suggest that the PVH plays a critical role in the modulation of ACTH and beta-endorphin release from the pituitary, and that CRF-containing pathways in the brain are involved in the mediation of autonomic responses.

The organization of noradrenergic pathways from the brainstem to the paraventricular and supraoptic nuclei in the rat

Axonal transport and immunohistochemical methods have been used to clarify the organization of pathways from noradrenergic and adrenergic cell groups in the brainstem to the paraventricular (PVH) and supraoptic (SO) nuclei of the hypothalamus. First, the location of such cells was determined with a combined retrograde tracer-immunofluorescence method. The fluorescent tracer, True Blue, was injected into the PVH or the SO, and sections through the brainstem were stained with anti-(rat) DBH, a specific marker for noradrenergic and adrenergic neurons. It was found that, after injections in the PVH, doubly labeled neurons were confined almost exclusively to 3 cell groups, the A1 region of the ventral medulla, which contained a majority of such cells, the A2 region in the dorsal vagal complex, and the locus coeruleus (A6 region). After injections centered in the SO an even greater proportion of doubly labeled cells were found in the A1 region, although some were also found in the A2 and A6 regions. The topography of doubly labeled cells indicates that these projections arise primarily from noradrenergic neurons, although adrenergic cells in both the C1 and the C2 groups probably contribute as well. Because well over 80% of the retrogradely labeled cells in these three regions were also DBH-positive, we next placed injections of [3H]amino acids into each of them in different groups of animals, and traced the course and distribution of the ascending (presumably DBH-positive) projections to the PVH and SO in the resulting autoradiograms. Injections centered in the A1 region labeled a substantial projection to most parts of the parvocellular division of the PVH, and was most dense in the dorsal and medial parts. In addition, terminal fields were labeled on those parts of the magnocellular division of the PVH, and of the SO, in which vasopressinergic cell bodies are concentrated. Injections centered in the A2 region also labeled a projection to the parvocellular division of the PVH that was topographically similar, but less dense, than that from the A1 region. In contrast, [3H]amino acid injections centered in the locus coeruleus labeled a moderately dense projection to the PVH that was limited to the medialmost part of the parvocellular division. Neither the A2 nor the A6 cell groups project to the magnocellular parts of PVH, or to the SO. The autoradiographic material, and additional double-labeling experiments, were used to identify and to characterize projections that interconnect the A1, A2 and A6 regions, as well as possible projections from these cell groups to the spinal cord. These results may be summarized as follows: a substantial projection from the nucleus of the solitary tract to the A1 region was identified, but this pathway does not arise from catecholaminergic neurons in the A2 cell group. DBH-stained cells in the A1 region project back to the dorsal vagal complex, as well as quite massively to the locus coeruleus (A6 region)...

Paraventricular Nucleus:A Site for the Integration of Neuroendocrine and Autonomic Mechanisms

We have summarized here recent evidence that clarifies the cellular organization and connections of the paraventricular nucleus of the hypothalamus (PVH) in the rat. The nucleus consists of a magnocellular division, with three distinct parts, and a parvocellular division with five distinct parts. Most neurons in the magnocellular division contain either oxytocin or vasopressin, and project to the posterior lobe of the pituitary gland. Separate cell populations centered in the parvocellular division give rise to projections to the median eminence, or to the brain stem and spinal cord including the intermediolateral column; some cells project both to the dorsal vagal complex and to the spinal cord. Cells with long descending projections may contain either oxytocin, vasopressin, somatostatin, or dopamine, although the biochemical specificity of most such neurons has not been determined. Noradrenergic fibers are found preferentially within those parts of the magnocellular division that are predominantly vasopressinergic. The parvocellular division is innervated by adrenergic as well as noradrenergic fibers from the brain stem, and by fibers from the dorsal vagal complex and the parabrachial nucleus. The bed nucleus of the stria terminalis and adjacent parts of the hypothalamus also innervate the PVH. The evidence indicates that subpopulations of neurons in the PVH are directly related to autonomic and neuroendocrine effector mechanisms, and suggest that the nucleus plays an important role in the regulation of visceral responses in the periphery and in the CNS itself.

A tissue-specific transcription factor containing a homeodomain specifies a pituitary phenotype

Multiple related cis-active elements required for cell-specific activation of the rat prolactin gene appear to bind a pituitary-specific positive transcription factor(s), referred to as Pit-1. DNA complementary to Pit-1 mRNA, cloned on the basis of specific binding to AT-rich cell-specific elements in the rat prolactin and growth hormone genes, encodes a 33 kd protein with significant similarity at its carboxyl terminus to the homeodomains encoded by Drosophila developmental genes. Pit-1 mRNA is expressed exclusively in the anterior pituitary gland in both somatotroph and lactotroph cell types, which produce growth hormone and prolactin, respectively. Pit-1 expression in heterologous cells (HeLa) selectively activates prolactin and growth hormone fusion gene expression, suggesting that Pit-1 is sufficient to confer a characteristic pituitary phenotype. The structure of Pit-1 and its recognition elements suggests that metazoan tissue phenotype is controlled by a family of transcription factors that bind to related cis-active elements and contain several highly conserved domains.

Organization of projections from the medial nucleus of the amygdala: a PHAL study in the rat.

The organization of axonal projections from the four recognized parts of the medial amygdalar nucleus (MEA) were characterized with the Phaesolus vulgaris leucoagglutinin (PHAL) method in male rats. The results indicate that the MEA consists of two major divisions, ventral and dorsal, and that the former may also consist of rostral and caudal regions. As a whole, the MEA generates centrifugal projections to several parts of the accessory and main olfactory sensory pathways, and projections to (a) several parts of the intrahippocampal circuit (ventrally); (b) the ventral striatum, ventral pallidum, and bed nuclei of the stria terminalis (BST) in the basal telencephaon; (c) many parts of the hypothalamus; (d) midline and medial parts of the thalamus; and (e) the periaqueductal gray, ventral tegmental area, and midbrain raphé. The dorsal division of the MEA (the posterodorsal part) is characterized by projections to the principal nucleus of the BST, and to the anteroventral periventricular, medial, and central parts of the medial preoptic, and ventral premammillary hypothalamic nuclei. These hypothalamic nuclei project heavily to neuroendocrine and autonomic‐related parts of the hypothalamic periventricular zone. The ventral division of the MEA (the anterodorsal, anteroventral, and posteroventral parts) is characterized by dense projections to the transverse and interfascicular nuclei of the BST, and to the lateral part of the medial preoptic, anterior hypothalamic, and ventromedial hypothalamic nuclei. However, dorsal regions of the ventral division provide rather dense inputs to the medial preoptic region and capsule of the ventromedial nucleus, whereas ventral regions of the ventral division preferentially innervate the anterior hypotha lamic, dorsomeclial, and ventral parts of the ventromedial nuclei. Functional evidence suggests that circuits associated with dorsal regions of the ventral division may deal with reproductive behavior, whereas circuits associated with ventral regions of the ventral division may deal preferentially with agonistic behavior.

Cerebral hemisphere regulation of motivated behavior

The goals of this article are to suggest a basic wiring diagram for the motor neural network that controls motivated behavior, and to provide a model for the organization of cerebral hemisphere inputs to this network. Cerebral projections mediate voluntary regulation of a behavior control column in the ventromedial upper brainstem that includes (from rostral to caudal) the medial preoptic, anterior hypothalamic, descending paraventricular, ventromedial, and premammillary nuclei, the mammillary body, and finally the substantia nigra and ventral tegmental area. The rostral segment of this column is involved in controlling ingestive (eating and drinking) and social (defensive and reproductive) behaviors, whereas the caudal segment is involved in controlling general exploratory or foraging behaviors (with locomotor and orienting components) that are required for obtaining any particular goal object. Virtually all parts of the cerebral hemispheres contribute to a triple descending projection - with cortical excitatory, striatal inhibitory, and pallidal disinhibitory components - to specific parts of the behavior control column. The functional dynamics of this circuitry remain to be established.

Organization of angiotensin II immunoreactive cells and fibers in the rat central nervous system. An immunohistochemical study.

The distribution of angiotensin II (AII)-immunoreactive cells and fibers was examined in adult male Sprague-Dawley rats with and without colchicine pretreatment. As seems to be the case for a number of other neuropeptides, AII is preferentially found in brain stem, hypothalamic, and limbic structures involved in the control of homeostatic functions. AII-stained cell bodies were most prominent in magnocellular parts of the paraventricular and supraoptic nuclei, and cells were also found in parvocellular parts of the former. Other hypothalamic nuclei containing cell bodies include the suprachiasmatic nucleus, the medial preoptic area, and perifornical parts of the lateral hypothalamic area. Of considerable interest was robust staining in several of the circumventricular organs, in particular the subfornical organ, where both cells and fibers were found. The results of water deprivation and nephrectomy suggest that this staining does not represent uptake of circulating peptide, but instead, represents AII-containing neural connections. In the thalamus, AII-stained cells were found in the paraventricular nucleus, the central medial nucleus, the nucleus reuniens, and rostral parts of the zona incerta. Two cell groups in the basal telencephalon, in the dorsal part of the bed nucleus of the stria terminalis and in the medial nucleus of the amygdala, lay at either end of an AII-stained pathway coursing through the stria terminalis. In the midbrain, immunoreactive cells were found in the interpeduncular and peripeduncular nuclei, and one pontine cell group was detected in the most lateral part of the lateral parabrachial nucleus. The only AII-stained cells in the medulla were in the nucleus of the solitary tract, near the margin of the area postrema. Fibers were found at all levels of the central nervous system, from the olfactory bulbs to the spinal cord, where terminal fields were observed in the substantia gelatinosa and in the intermediolateral cell column. Longitudinally oriented fibers were present throughout the periventricular fiber system and in the medial forebrain bundle, including its caudal extension in ventrolateral parts of the brain stem. It is suggested that, at many different levels, AII serves as both a hormone and neurotransmitter for fluid balance.

Topography of projections from amygdala to bed nuclei of the stria terminalis.

A collection of 125 PHAL experiments in the rat has been analyzed to characterize the organization of projections from each amygdalar cell group (except the nucleus of the lateral olfactory tract) to the bed nuclei of the stria terminalis, which surround the crossing of the anterior commissure. The results suggest three organizing principles of these connections. First, the central nucleus, and certain other amygdalar cell groups associated with the main olfactory system, innervate preferentially various parts of the lateral and medial halves of the bed nuclear anterior division, and these projections travel via both the stria terminalis and ansa peduncularis (ventral pathway). Second, in contrast, the medial nucleus, and the rest of the amygdalar cell groups associated with the accessory and main olfactory systems innervate preferentially the posterior division, and the medial half of the anterior division, of the bed nuclei. And third, the lateral and anterior basolateral nuclei of the amygdala (associated with the frontotemporal association cortical system) do not project significantly to the bed nuclei. For comparison, inputs to the bed nuclei from the ventral subiculum, infralimbic area, and endopiriform nucleus are also described. The functional significance of these projections is discussed with reference to what is known about the output of the bed nuclei.

The neuronal mineralocorticoid eeceptor as a mediator of glucocorticoid response

The cloning of the mineralocorticoid receptor (MR) and the glucocorticoid receptor (GR) cDNAs provides a basis for understanding the actions of glucocorticoids in the central nervous system. Structural evidence is presented for the identity of the type I corticosteroid binding site as the MR expressed in the brain. This identification is supported by the anatomical distribution of MR mRNA, determined by in situ hybridization histochemistry, which parallels the steroid autoradiographic localization of the type I sites. An in vitro assay for MR and GR function demonstrates that these receptors respond to different levels of glucocorticoid, suggesting that together they confer a larger dynamic range of sensitivity to this hormone. These studies lead to a new hypothesis for glucocorticoid action in the central nervous system.

Connections of the rat lateral septal complex

The organization of lateral septal connections has been re-examined with respect to its newly defined subdivisions, using anterograde (PHAL) and retrograde (fluorogold) axonal tracer methods. The results confirm that progressively more ventral transverse bands in the hippocampus (defined by the orientation of the trisynaptic circuit) innervate progressively more ventral, transversely oriented sheets in the lateral septum. In addition, hippocampal field CA3 projects selectively to the caudal part of the lateral septal nucleus, which occupies topologically lateral regions of the transverse sheets, whereas field CA1 and the subiculum project selectively to the rostral and ventral parts of the lateral septal nucleus, which occupy topologically medial regions of the transverse sheets. Finally, the evidence suggests that progressively more ventral hippocampal bands innervate progressively thicker lateral septal sheets. In contrast, ascending inputs to the lateral septum appear to define at least 20 vertically oriented bands or subdivisions arranged orthogonal to the hippocampal input (Risold, P.Y. and Swanson, L.W., Chemoarchitecture of the rat lateral septal nucleus, Brain Res. Rev., 24 (1997) 91-113). Hypothalamic nuclei forming parts of behavior-specific subsystems share bidirectional connections with specific subdivisions of the lateral septal nucleus (especially the rostral part), suggesting that specific domains in the hippocampus may influence specific hypothalamic behavioral systems. In contrast, the caudal part of the lateral septal nucleus projects to the lateral hypothalamus and to the supramammillary nucleus, which projects back to the hippocampus and receives its major inputs from brainstem cell groups thought to regulate behavioral state. The neural system mediating defensive behavior shows these features rather clearly, and what is known about its organization is discussed in some detail.