Regulation of the galanin system by repeated electroconvulsive seizures in mice

Even though induction of seizures by electroconvulsive stimulation (ECS) is a treatment widely used for major depression in humans, the working mechanism of ECS remains uncertain. The antiepileptic effect of ECS has been suggested to be involved in mediating the therapeutic effect of ECS. The neuropeptide galanin exerts antiepileptic and antidepressant-like effects and has also been implicated in the pathophysiology of depression. To explore a potential role of galanin in working mechanisms of ECS, the present study examined effects of repeated ECS on the galanin system using QRT-PCR, in situ hybridization, and [(125) I]galanin receptor binding. ECS was administered to adult mice daily for 14 days, and this paradigm was confirmed to exert antidepressant-like effect in the tail suspension test. Prominent increases in galanin gene expression were found in several brain regions involved in regulation of epileptic activity and depression, including the piriform cortex, hippocampal dentate gyrus, and amygdala. Likewise, GalR2 gene expression was up-regulated in both the central and the medial amygdala, whereas GalR1 gene expression showed a modest down-regulation in the medial amygdala. [(125) I]galanin receptor binding in the piriform cortex, hippocampus, and amygdala was found to be significantly down-regulated. These data show that the galanin system is regulated by repeated ECS in a number of brain regions implicated in seizure regulation and depression. These changes may play a role in the therapeutic effect of ECS.     

Distribution and differential regulation of galanin receptor subtypes in rat brain: effects of seizure activity

Galanin, acting at the GalR1-3 subtypes of galanin receptors, is involved in the regulation of cognition, mood, feeding, seizure activity and pain. The understanding of galanin's effects in molecular and cellular terms has been hampered by the lack of receptor subtype selective ligands and antibodies. Previous in situ hybridization data showed that GalR1 and GalR2 receptors are abundant in the rat brain, while the distribution of GalR3 is contradictory and most studies demonstrated a low expression levels in the rat brain. The distribution of galanin receptor subtypes at protein level is unknown. In the present study, we report the regional distribution of the galanin receptors: GalR1 and non-GalR1 receptors, using a recently synthesized high affinity GalR2/3 selective ligand, galanin (2-11), and galanin (1-29), as competitors, in saturating (125)I-galanin membrane binding assay. We show that paraventricular nucleus (PVN) express predominantly GalR1, whereas areas like the dorsal raphe nucleus (DRN), hippocampus and amygdala express both the GalR1 and non-GalR1 receptors. We speculate that the GalR2/3 binding sites detected by galanin (2-11) binding in our study probably represent mostly GalR2 receptors. In addition, we show regionally specific and subtype specific regulation of galanin receptors. Status epilepticus (SE), known to deplete galanin from axonal projections of locus coeruleus and septum/diagonal band neurons in the hippocampus and to induce galanin expression in a subset of hippocampal cells, down regulates GalR2 receptor mRNA and proteins by 30% without altering the GalR1 receptors.     

Changes in galanin and GalR1 gene expression in discrete brain regions after transient occlusion of the middle cerebral artery in female rats

Injury to neurons results in up-regulation of galanin in some central and peripheral systems, and it has been suggested that this neuropeptide may play a protective and trophic role, primarily mediated by galanin receptor 2 (GalR2). The objective of the present study was to investigate galanin, GalR1, GalR2 and GalR3 gene expression in the female rat brain 7 days after a 60-min unilateral occlusion of the middle cerebral artery followed by reperfusion. Quantitative real-time PCR was employed in punch-biopsies from the locus coeruleus, somatosensory cortex and dorsal hippocampal formation, including sham-operated rats as controls. Galanin gene expression showed a ～2.5-fold increase and GalR1 a ～1.5-fold increase in the locus coeruleus of the ischemic hemisphere compared to the control side. Furthermore, the GalR1 mRNA levels decreased by 35% in somatosensory cortex of the ischemic hemisphere. Immunohistochemical analysis indicated a depletion of galanin from cell bodies and dendrites in the locus coeruleus after middle cerebral artery occlusion. The present results suggest that a stroke-induced forebrain lesion up-regulates synthesis of galanin and GalR1 in the locus coeruleus, a noradrenergic cell group projecting to many forebrain areas, including cortex and the hippocampal formation. These results support the notion that galanin may play a role in the response of the central nervous system to injury.     

The brain galanin receptors: targets for novel antidepressant drugs

Our present view that the mood disorders involve dysfunction of monoaminergic system is a result of important clinical and preclinical observations over the past 40 years. The therapeutic efficacy of drugs such as the tricyclic antidepressants (TCAs), monoamine oxidase inhibitors, selective serotonin reuptake inhibitors (SSRIs) and lately of SNRIs (serotonin and norepinephrine reuptake inhibitors) helped to shape our view that mood regulation involves the monoaminergic systems in some way. It is thus little surprising when the neuropeptide, galanin, is discovered to coexist with norepinephrine (NE) in locus coeruleus (LC) neurons and with serotonin (5-HT) in the dorsal raphe nucleus (DRN) neurons, a link between galanin mediated signaling and mood regulation is sought. Galanin receptors are expressed in brain structures that are involved in the regulation of mood such as frontal cortex, amygdala, hypothalamus, LC, DRN and hippocampus. It is almost an accident of research fate that the potent effects of galanin on cognitive performance and seizure threshold have led galanin research to focus on the hippocampus where the neuropeptide is present in cholinergic and noradrenergic afferents and where the receptor density is much lower than in the monoaminergic nuclei. Hopefully it is not too late to report on the recent inroads into the roles of galanin and of galanin receptor subtypes 2 and 3 (GalR2 and GalR3) in mood regulation in animal models as well as in human patients with major depression. A body of existing data suggests that GalR2 signaling leads to antidepressant-like, anticonvulsant and neurogenesis-promoting effects, a spectrum of activities that are commonly associated with efficacious antidepressants. Similarly, GalR3 antagonists exhibit anxiolytic and antidepressant-like activity, another clinically useful combination for the treatment of mood disorders. Since both GalR2 and GalR3 are G-protein coupled receptors (GPCRs), a favorite target class for drug development, we believe that the pace of developing galaninergic antidepressants will increase significantly from now on.     

Raphe projections to the locus coeruleus in the rat

Afferent projections to the locus coeruleus from the various raphe nuclei, particularly of the midbrain (nuclei raphe dorsalis and medianus) and pons (nuclei raphe pontis and magnus), have been studied in the rat by retrograde transport methods using horseradish peroxidase (HRP). The locus coeruleus, in both its dorsomedial and ventrolateral divisions, and in its various anterior-posterior components, were injected with 0.05 μl of horseradish peroxidase following which various structures of the brainstem, particularly the raphe nuclei, were examined for HRP reactive cells. It was found that injections in most components of the locus coeruleus were associated with HRP positive cells in varying degrees of density in the nuclei raphe dorsalis, medianus, pontis, and magnus, with considerably sparser labelling in the anterior aspects of the medullary raphe nuclei pallidus and obscurus. Labelled cells were also seen in the nuclei of the solitary tract, contralateral locus coeruleus, lateral reticular areas of the pons and midbrain, nuclei pontis oralis and caudalis, vestibular nuclei, mesencephalic nucleus of the trigeminal nerve, fastigial nuclei of cerebellum and medial parabrachial nuclei. These data, showing widespread innervation of the locus coeruleus from all raphe nuclei, as well as many other brainstem areas, in the rat support the general view of heavy innervation of the locus coeruleus from both extra-raphe and raphe nulcei. These latter raphe projections, probably serotonergic in nature, provide anatomical support for the various experiments indicating considerable regulation of locus coeruleus activities, such as phasic events of REM sleep, among others, by most of the raphe nuclei. Thus, various activities of the locus coeruleus could be modulated or regulated by widespread projections from most raphe nuclei as well as several other regions of the brainstem.     

G protein-coupled galanin receptor distribution in the rat central nervous system

The action of galanin in the central nervous system is mediated by at least three galanin receptor subtypes (GalR1, GalR2 and GalR3) which belong to the family of G protein-coupled receptors. GalR1 and GalR2 are coupled to G(i/o) proteins, although the latter may also be coupled to G(q/11) proteins. The aim of the present study was to identify the anatomical distribution and quantify the density of GalRs coupled to G proteins. The galanin (10(-6) M) stimulated guanosine 5'-(gamma-[35S] thio)triphosphate binding assay was used in tissue sections from the rat brain. Maximal percentages of stimulation over basal levels were found in the anterior olfactory nucleus and in the lateral olfactory tract nucleus ( approximately 54%). High levels of stimulation were recorded in diverse hypothalamic nuclei (16-28%), in the amygdala (central amygdaloid nucleus, 40%), in the spinal trigeminal tract (23%) and in layers 1-2 of the spinal cord (26%). Moderate binding stimulation (5-13%) was observed in thalamus, substantia nigra pars compacta, parabrachial nucleus, locus coeruleus and dorsal raphe nucleus. The lowest stimulation induced by galanin was recorded in diverse areas of the cortex, striatum, hippocampus and substantia nigra pars reticulata. The results show an anatomical distribution similar to that described for GalR1. However, in diverse brain areas, in which a high density of these receptors has previously been reported, only a moderate coupling to G proteins was found. These findings would suggest that the efficacy of galanin to induce an effective coupling of its receptors to G proteins could be different depending on the brain area.     

Influence of chronic administration of antidepressant drugs on mRNA for galanin, galanin receptors, and tyrosine hydroxylase in catecholaminergic and serotonergic cell-body regions in rat brain

Activity of locus coeruleus (LC) neurons and release of the peptide galanin (GAL), which is colocalized with norepinephrine (NE) in LC neurons, has been implicated in depression and, conversely, in antidepressant action. The present study examined the influence of chronic administration (for 14days, via subcutaneously-implanted minipump) of antidepressant (AD) drugs representing three different classes (tricyclic [desipramine], selective serotonin reuptake inhibitor [SSRI] [paroxetine], and monoamine oxidase inhibitor [MAOI] [phenelzine]) on mRNA for GAL, GAL receptors (GalR1, GalR2, and GalR3), and tyrosine hydroxylase (TH), the rate-limiting enzyme for NE synthesis, in four brain regions--LC, A1/C1, dorsal raphe (DRN), and ventral tegmentum (VTA) of rats. Consistent with previous findings that chronic administration of AD drugs decreases activity of LC neurons, administration of AD drugs reduced mRNA for both GAL and TH in LC neurons. GAL and TH mRNA in LC neurons was highly correlated. AD drugs also reduced GAL and TH mRNA in A1/C1 and VTA but effects were smaller than in LC. The largest change in mRNA for GAL receptors produced by AD administration was to decrease mRNA for GalR2 receptors in the VTA region. Also, mRNA for GalR2 and GalR3 receptors was significantly (positively) correlated in all three predominantly catecholaminergic brain regions (LC, A1/C1, and VTA). Relative to these three brain regions, unique effects were seen in the DRN region, with the SSRI elevating GAL mRNA and with mRNA for GalR1 and GalR3 being highly correlated in this brain region. The findings show that chronic administration of AD drugs, which produces effective antidepressant action, results in changes in mRNA for GAL, GAL receptors, and TH in brain regions that likely participate in depression and antidepressant effects.     

Sources of cortical, hypothalamic and spinal serotoninergic projections: topical organization in the dorsal Raphe nucleus]

Distribution of primuline, fast blue, fluoro-gold and nuclear yellow-labelled monoamine-containing cells in periventricular gray and dorsolateral tegmentum (including locus coeruleus) was studied in the rat after injection of these fluorochromes into the frontal cortex, hypothalamus and spinal cord. Combination of monoamine fluorescence method and retrograde cell-labelling was used. Two big groups of serotonin-positive cells projecting into the upper thoracic spinal segments were found in dorsomedial zone of the dorsal raphe. Part of these units also had divergent axon projections to the frontal cortex. Such cellular arrangement allows a supposition that analgetic effects of dorsal raphe stimulation can be partially based on the direct participation of this structure in the descending control at the spinal level. Neurones, sources of cortical projections are intermingled with the cells projecting to the hypothalamus but some topical differentiation can be distinguished. Neurotransmitter and neuroregulatory roles of separate cortical and hypothalamic projections of serotonin-containing neurons of the dorsal raphe cells is discussed.     

Afferent projections to affective attack sites in cat hypothalamus

Quiet biting attack by a cat on a rat was elicited by electrical stimulation of sites in the cat's lateral hypothalamus.Horseradish peroxidase was deposited at the attack sites. Cells containing reaction products were found in gyrus proreus, anterior and central medial amygdaloid nuclei, lateral and medial preoptic areas, substantia innominata, the bed nuclei of stria terminalis, and anterior commissure. The dorsomedial area of the hypothalamus, paraventricular nucleus, suprammamillary region, and posterior hypothalamic area also contained reactive cells. In the midbrain the ventral tegmental area of Tsai, the dorsal and superior central nuclei of the raphe, central gray matter and interpeduncular nucleus were regions with reactive cells. In the pontine region, the locus coeruleus, parabrachial nuclei, nucleus of the lateral lemniscus, and the dorsal tegmental nucleus of Gudden all had reactive cells.There are many structures which send afferent projections to quiet attack sites located in the hypothalamus and the pontine tegmentum. The commonality of afferents to attack sites lends credence to the notion that a complex, distributed, interactive network underlies the neural basis of attack behavior.     

Efferents and afferents of the ventral tegmental-A10 region studied after local injection of [H]leucine and horseradish peroxidase

The efferents and the afferents of the VMT-A10 region were studied by using anterograde ([³H]leucine) and retrograde (HRP) tracing techniques. In order to produce very small injections in various parts of the VMT-A10 region, a slow diffusion technique for [³H]leucine labelling and a microiontophoretic injection for horseradish peroxidase labelling were developed. According to the histochemical and biochemical data, the [³H]leucine anterograde results were separated into three main types of projections.

(1) Projections to regions rich in DA terminals. These projections certainly correspond to the efferents of the dopaminergic A10 neurones. According to various injection sites, we have been able to identify mesolimbic projections originating from the VMT-A10, pars medialis and mesostriatal-mesolimbic projections originating from the VMT-A10, pars lateralis.

The mesolimbic projections include the prefrontal cortex, the medial part of the lateral septum, the interstitial nucleus of the stria terminalis, the accumbens nucleus and the olfactory tubercle. The mesostriatal-mesolimbic projections include the anteromedial part of the caudate nucleus, the cingular cortex, the entorhinal cortex, the amygdaloid complex, the accumbens nucleus, the olfactory tubercle and the piriform cortex to a lesser extent.

(2) Projections to regions suspected of containing DA terminals. These ascending and descending projections which could represent the dopaminergic efferents of the VMT-A10 neurones have been demonstrated. Ascending projections originating either from the VMT-A10 pars medialis or pars lateralis region were found in the claustrum, the nucleus of the tractus diagonalis, the olfactory nuclei, the lateral habenula, the medial hypothalamus and the median eminence. The projections observed in the medial hypothalamus included the periventricular region, the arcuate nucleus, the ventral part of the ventromedial nucleus and the dorsomedial nucleus. The labelling of the anteromedial part of the dorsal hippocampus appeared to originate from the VMT-A10, pars posterior. The projections to the medial hypothalamus, median eminence and hippocampus may have a great functional significance, but further proof of their dopaminergic nature is needed. Descending projections were found ipsilateally to the dorsal raphe and to the cerebellum, and bilaterally to the locus coeruleus. The projections to the cerebellum are distributed to the nuclei interpositus and dentatus and to the Purkinje cell layer and granular layer of the cortex. These results raise the problem of descending dopaminergic projections from the A10 neurones.

(3) Projections to regions not known to contain DA terminals. Anterior projections were found ipsilaterally to the supraoptic nucleus and bilaterally to the anterodorsal thalamic nucleus. Posterior projections were traced ipsilaterally to the limbic midbrain area, including the median raphe, the ventral and dorsal tegmental nucleus and the central gray.

The horseradish peroxidase experiment supplied some clues as to the posterior afferents of the VMT-A10 region. Some labelled cells were found ipsilaterally in the substantia nigra, the medain raphe and the ventral tegmental nucleus. Numerous cells were labelled ipsilaterally in the dorsal raphe nucleus, and nuclei interpositus and dentatus of the cerebellum, and contralaterally in the locus coeruleus. These structures are likely to play an important role in the modulation of the activity of VMT-A10 neurones.

The results of [³H]leucine and HRP experiments permitted us to demonstrate reciprocal connections between VMT-A10 region and anterior raphe nuclei, locus coeruleus and cerebellum.

Septal complex of the telencephalon of the lizard Podarcis hispanica. II. afferent connections

The afferent connections to the septal complex were studied in the lizard Podarcis hispanica (Lacertidae) by means of a combination of retrograde and anterograde tracing. The results of these experiments allow us to classify the septal nuclei into three main divisions. The central septal division (anterior, lateral, dorsolateral, ventrolateral, and medial septal nuclei plus the nucleus of the posterior pallial commissure) receives a massive, topographically organized, cortical projection (medial, dorsal, and ventral areas) and widespread afferents from the tuberomammillary hypothalamus and the basal telencephalon. Moreover, it receives discrete projections from the dorsomedial anterior thalamus, the ventral tegmentum, the midbrain raphe, and the locus coeruleus. The ventromedial septal division (ventromedial septal nucleus) receives a massive projection from the anterior hypothalamus, dense serotonergic innervation, and a faint amygdalohypothalamic projection, but it is devoid of direct cortical input. The midline septal division (nucleus septalis impar and dorsal septal nucleus) receives a nontopographic cortical projection (dorsomedial and dorsal cortices) and afferents from the preoptic hypothalamus, the dorsomedial anterior thalamus, the midbrain central gray, and the reptilian A8 nucleus/substantia nigra. Our results indicate that the cortex provides a physiologically complex, massive input to the septum that terminates over the whole dendritic tree of septal cells. In contrast, most of the ascending afferents make axosomatic contacts by means of pericellular nests. The chemical nature of the main septal afferents and the comparative implications of the available hodological data on the organization of the septal complex of tetrapod vertebrates are discussed. J. Comp. Neurol. 383:489-511, 1997. © 1997 Wiley-Liss, Inc.     

Increased corticotropin-releasing hormone immunoreactivity in monoamine-containing pontine nuclei of depressed suicide men

A number of clinical investigations and postmortem brain studies have provided evidence that excessive corticotropin-releasing hormone (CRH) secretion and neurotransmission is involved in the pathophysiology of depressive illness, and several studies have suggested that the hyperactivity in CRH neurotransmission extends beyond the hypothalamus involving several extra-hypothalamic brain regions. The present study was designed to test the hypothesis that CRH levels are increased in specific brainstem regions of suicide victims with a diagnosis of major depression. Frozen tissue sections of the pons containing the locus coeruleus and caudal raphe nuclei from 11 matched pairs of depressed suicide and control male subjects were processed for radioimmunocytochemistry using a primary antiserum to CRH and a ([125])I-IgG secondary antibody. The optical density corresponding to the level of CRH-immunoreactivity (IR) was quantified in specific pontine regions from the film autoradiographic images. The level of CRH-IR was increased by 30% in the locus coeruleus, 39% in the median raphe and 45% in the caudal dorsal raphe in the depressed suicide subjects compared to controls. No difference in CRH-IR was found in the dorsal tegmentum or medial parabrachial nucleus between the subject groups. These findings reveal that CRH-IR levels are specifically increased in norepinephrine- and serotonin-containing pontine nuclei of depressed suicide men, and thus they are consistent with the hypothesis that CRH neurotransmission is elevated in extra-hypothalamic brain regions of depressed subjects.     

Selective prefrontal cortical projections to the region of the locus coeruleus and raphe nuclei in the rhesus monkey

Descending cortical projections to the region of the noradrenergic locus coeruleus (LC) and the serotonergic dorsal and central superior raphe nuclei were analyzed in the rhesus monkey using anterograde labeling techniques. HRP pellets or tritiated leucine were injected into one of 7 cortical areas: the dorsolateral prefrontal cortex, the dorsomedial prefrontal cortex, the orbital prefrontal cortex, the parietal association cortex, somatosensory cortex, the anterior portion of the inferior temporal gyrus, and the posterior portion of the inferior temporal gyrus. Anterogradely labeled fibers were found in and adjacent to the LC and raphe nuclei only following the dorsolateral and dorsomedial prefrontal cortical injections. Terminal labeling was densest at rostral levels of the LC, particularly in the area directly medial to the nucleus. Labeled fibers could not be followed beyond caudal levels of the LC. The projections to the contralateral LC and raphe nuclei were similar to, but less dense than that to the ipsilateral region. Injections into cortical areas other than the dorsal prefrontal cortex resulted in anterograde labeling of the pontine nuclei or pyramids, but not the LC/raphe region. These data, in conjunction with studies in the rat, suggest that the dorsal prefrontal cortex may be the only cortical area to have direct influence on the LC and raphe nuclei and secondary influence on the monoaminergic innervation of large areas of cerebral cortex.     

Connections of the ventral telencephalon (subpallium) in the zebrafish (Danio rerio)

Connections of the medial precommissural subpallial ventral telencephalon, i.e., dorsal (Vd, interpreted as part of striatum) and ventral (Vv, interpreted as part of septum) nuclei of area ventralis telencephali, were studied in the zebrafish (Danio rerio) using two tracer substances (DiI or biocytin). The following major afferent nuclei to Vd/Vv were identified: medial and posterior pallial zones of dorsal telencephalic area, and the subpallial supracommissural and postcommissural nuclei of the ventral telencephalic area, the olfactory bulb, dorsal entopeduncular, anterior and posterior parvocellular preoptic and suprachiasmatic nuclei, anterior, dorsal and central posterior dorsal thalamic, as well as rostrolateral nuclei, periventricular nucleus of the posterior tuberculum, posterior tuberal nucleus, various tuberal hypothalamic nuclei, dorsal tegmental nucleus, superior reticular nucleus, locus coeruleus, and superior raphe nucleus. Efferent projections of the ventral telencephalon terminate in the supracommissural nucleus of area ventralis telencephali, the posterior zone of area dorsalis telencephali, habenula, periventricular pretectum, paracommissural nucleus, posterior dorsal thalamus, preoptic region, midline posterior tuberculum (especially the area dorsal to the posterior tuberal nucleus), tuberal (midline) hypothalamus and interpeduncular nucleus. Strong reciprocal interconnections likely exist between septum and preoptic region/midline hypothalamus and between striatum and dorsal thalamus (dopaminergic) posterior tuberculum. Regarding ascending activating/modulatory systems, the pallium shares with the subpallium inputs from the (noradrenergic) locus coeruleus, and the (serotoninergic) superior raphe, while the subpallium additionally receives such inputs from the (dopaminergic) posterior tuberculum, the (putative cholinergic) superior reticular nucleus, and the (putative histaminergic) caudal hypothamalic zone.     

Connectivity of the lobus parolfactorius of the domestic chicken (Gallus domesticus): An anterograde and retrograde pathway tracing study

In 1–week-old domestic chicks, the connectivity of the lobus parolfactorius (LPO), part of the avian basal ganglia, was investigated using Phaseolus vulgaris leucoagglutinin and horseradish peroxidase for anterograde and retrograde pathway tracing, respectively. Tyrosine hydroxylase immunocytochemistry was applied in combination with Phaseolus lectin to assess the overlap between LPO efferents and diencephalic and meseticephalic catecholamine centres. Anterograde projections from LPO were detected in the hyperstriatum, neostriatum, and paleostriatum. Intranuclear connections were also apparent within the LPO. Descending LPO efferents innervated the lateral mammillary and intramedial nuclei and the dorsomedial thalamic complex. Fibres from LPO were observed in the tectal gray, substantia nigra, area ventralis tegmentalis of Tsai, and the adjacent nucleus mesencephalicus profundus. Further caudally, projections from LPO reached the nucleus papillioformis, locus coeruleus, and subcoeruleus ventralis. LPO efferents were coextensive with tyrosine hydroxylase-positive cells in the nuclei mamillaris lateralis and intramedialis of the hypothalamus, area ventralis tegmentalis, substantia nigra, locus coeruleus, and subcoeruleus ventralis of mesencephalic and pontine tegmentum. Close contacts between LPO fibres and catecholamine cells were visible in the nigra and the area ventralis tegmentalis. Retrograde labelling from LPO was found in the archistriatum, dorsomedial thalamic complex, nuclei lateralis anterior and superficialis parvicellularis thalami, substantia nigra, central gray, area ventralis tegmentalis of Tsai, and locus coeruleus and in cells dorsal to the decussation of brachium conjunctivum. Reciprocal connections were verified between the LPO and the following areas: dorsomedial thalamic complex, central gray, substantia nigra, area ventralis of Tsai, and locus coeruleus. © 1994 Wiley-Liss, Inc.     

Organization of galanin-immunoreactive inputs to the paraventricular nucleus with special reference to their relationship to catecholaminergic afferents

Immunohistochemical and axonal transport techniques were used to characterize the origin and distribution of galanin-immunoreactive inputs to the paraventricular (PVH) and supraoptic (SO) nuclei of the hypothalamus in the rat. In the parvicellular division of the PVH, the most prominent inputs were confined to the anterior and periventricular parts of the nucleus rostrally and the dorsal and ventral medial subdivisions caudally; the galaninergic inputs to the magnocellular division of PVH and SO were very sparse and were preferentially distributed to regions containing predominantly oxytocinergic neurons. A combined retrograde transport-immunohistochemical method was employed to identify sources of these projections. Galanin immunoreactivity was found to coexist with dopamine-beta-hydroxylase (DBH) immunoreactivity in subsets of retrogradely labeled neurons of the A1 and A6 (locus coeruleus) catecholamine cell groups; no evidence was adduced for the presence of galanin in adrenergic (i.e., phenylethanolamine-N-methyltransferase-positive) neurons that project to the PVH. Apart from minor contributions from the mesencephalic raphe nuclei, no other brainstem cell groups contributed to the galaninergic innervation of the PVH. In the forebrain, the most prominent grouping of doubly labeled cells was centered in the rostral part of the dorsomedial nucleus of the hypothalamus (DMH), though significant numbers were also found in the lateral hypothalamic area, the arcuate nucleus, and the medial preoptic area. In experiments designed to define the subnuclear specificity of some galanin-containing inputs to the PVH, iontophoretic deposits of the anterogradely transported plant lectin, Phaseolus vulgaris-leucoagglutinin (PHA-L), were placed in the A1 and A6 cell groups and in the DMH. Sections through the PVH were prepared so as to allow colocalization of anterogradely transported PHA-L and galanin immunoreactivity in individual fibers and varicosities. Consistent with the retrograde transport data, the greatest degree of galanin-PHA-L correspondence was seen after lectin deposits in the DMH, and over 80% of the doubly labeled varicosities were confined to the anterior, periventricular, and medial parvicellular subdivisions of the nucleus. The galanin-containing projection from the locus coeruleus was most circumscribed, with the vast majority of doubly labeled varicosities confined to the periventricular and adjoining aspects of the anterior and medial parvicellular subdivisions.(ABSTRACT TRUNCATED AT 400 WORDS)     

The brainstem origin of monoaminergic projections to the spinal cord of the North American opossum: A study using fluorescent tracers and fluorescence histochemistry

The retrograde transport of fluorescent markers has been combined with the glyoxylic acid and Falck-Hillarp techniques to identify the origin of monoamine axons within the spinal cord of the North American opossum. Catecholamine axons arise from neurons located within the ventrolateral medulla, dorsal to the superior olivary complex, within the dorsolateral and rostrolateral pons and within the periventricular nuclei of the hypothalamus. Such neurons are most numerous within the dorsolateral pons where they are found dorsal and lateral to the motor trigeminal nucleus, within the nucleus locus coeruleus pars alpha and adjacent reticular formation as well as within the ventral part of the nucleus locus coeruleus. Neurons containing the fluorescent marker and catecholamines were interspersed with others containing only the injected marker with the possible exception of the nucleus locus coeruleus. Spinal axons of the indoleamine type arise from neurons within the nuclei pallidus, obscurus and magnus raphe, the nucleus reticularis gigantocellularis, the nucleus reticularis gigantocellularis pars ventralis, the nucleus reticularis pontis pars ventralis and the nucleus dorsalis raphe. The latter nucleus only innervates rostral cervical levels. Most of the above areas also contain many non-indoleamine neurons which were labelled by the injected marker. This was particularly true of the nucleus magnus raphe and the adjacent nucleus reticularis points pars ventralis after injections of fluorescent markers into the superficial dorsal horn.     

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

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