Brainstem influences on transmission of somatosensory information in the spinocervicothalamic pathway

The inhibition of somatosensory responses of lateral cervical nucleus neurons resulting from stimulation of the brainstem has been investigated. Single unit extracellular recordings were obtained from neurons in the lateral cervical nucleus of chloralose-anesthetized cats. Electrical stimulation of the periaqueductal gray, nucleus raphe magnus, nucleus cuneiformis, and nuclei reticularis gigantocellularis and magnocellularis was found to be very effective in inhibiting the responses of lateral cervical nucleus neurons evoked by electrical or tactile stimulation of the skin. Additional experiments were performed to determine whether the inhibitory effects were mediated in the spinal cord dorsal horn or in the lateral cervical nucleus. These experiments which examined the effect of brainstem stimulation on the responses induced by stimulation of the dorsolateral funiculus or on the antidromic latency of activation of lateral cervical nucleus neurons from thalamus, revealed that most and possibly all the inhibition could be accounted for by an action on the spinal cord. These results are consistent with other studies showing that spinocervical tract cells in the spinal cord can be inhibited by stimulation of the same brainstem regions.     

Three bulbospinal pathways from the rostral medulla of the cat: an autoradiographic study of pain modulating systems

Small amounts of ³H-leucine were injected into discrete regions in the rostral medulla of the cat. Descending projections from these sites were studied with autoradiographic methods. On the basis of differential projections to the medulla and spinal cord, three distinct regions were delineated. Nucleus reticularis gigantocellularis (Rgc), located dorsally in the medullary reticular formation, projects primarily to “motor” related sites, including cranial motor nuclei VI, VII, XII, nucleus intercalatus, and a part of the ipsilateral medial accessory olive. The projection to the spinal cord is primarily via the ipsilateral ventrolateral and contralateral ventral funiculi. The Rgc terminal field is in lamina VII and VIII ipsilateral and lamina VIII contralateral to the injection site. In contrast, nucleus raphe magnus, (NRM) located ventrally, in the midline of the rostral medulla projects primarily to structures with known nociceptive and/or visceral afferent input. These sites include the solitary nucleus, the dorsal motor nucleus (X) and the marginal and gelatinous layers of the spinal trigeminal nucleus caudalis. The projection to the spinal cord is bilateral, via the dorsolateral funiculus. Terminal fields are found in the marginal zone and the substantia gelatinosa of the dorsal horn, and more deeply in lamina V, medial VI and VII. Nucleus reticularis magnocellularis (Rmc), located lateral to NRM and ventral to Rgc, has an overlapping projection with NRM, but the projection is ipsilateral. This difference between Rmc and Rgc is correlated with cytoarchitectural features of the two regions. The possibility that the raphe-spinal pathway in the DLF mediates opiate and brain stimulation-produced analgesia is discussed.     

Raphespinal projections in the north american opossum: Evidence for connectional heterogeneity

Retrograde transport studies revealed that the nuclei pallidus, obscurus, and magnus raphae as well as the adjacent reticular formation innervate the spinal cord in the opossum. HRP-lesion experiments showed that a relatively large number of neurons within the nucleus obscurus raphae and closely adjacent areas of the nucleus reticularis gigantocellularis project through the ventrolateral white matter and that many cells within the nucleus magnus raphae, the nucleus reticularis gigantocellularis pars ventralis, and the nucleus reticularis pontis pars ventralis contribute axons to the dorsal half of the lateral funiculi. Neurons within the rostral pole of the nucleus magnus raphae and the adjacent nucleus reticularis pontis pars ventralis may project exclusively through the latter route. Each of the above-mentioned raphe and reticular nuclei contain nonindolaminergic as well as indolaminergic neurons (Crutcher and Humbertson, 1978). When True-Blue was injected into the spinal cord and the brain processed for monoamine histofluorescence evidence for True-Blue was found in neurons of both types. Injections of ³H-leucine centered within the nuclei pallidus and obscurus raphae and/or the closely adjacent nucleus reticularis gigantocellularis labeled axons within autonomic nuclei and laminae IV-X. Labeled axons were particularly numerous within the intermediolateral cell column and within laminae IX and X. Injections of the caudoventral part of the nucleus magnus raphae or the adjacent nucleus reticualris gigantocellularis pars ventralis labeled axons in the same areas as well as within laminae I-III. When the injection was placed within the rostal part of the nucleus magnus raphae or the adjacent nucleus reticularis pontis pars ventralis axons were labeled within laminae I-III and external zones of laminae IV-VII, but not within lamina IX. The immunohistofluorescence method revealed evidence for indolaminergic axons in each of the spinal areas labeled by injections of ³H-leucine into the raphe and adjacent reticular formation. They were particularly abundant within the intermediolateral cell column and within laminae IX and X. These data indicate that raphe spinal systems are chemically and connectionally heterogeneous.     

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.     

Spinal projections from the lower brain stem in the cat as demonstrated by the horseradish peroxidase technique. I. Origins of the reticulospinal tracts and their funicular trajectories

Using a retrograde tracer technique with horseradish peroxidase (HRP) attempts were made to determine the origins of reticulospinal tracts and their funicular trajectories. Reticulospinal tracts originating from the mesencephalic reticular formation (RF) were composed of: (1) descending projections arising from the cluster of cells located just lateral to the periaqueductal gray that course in the anterior funiculus (AF) and ventral part of the lateral funiculus (LF) with ipsilateral predominance; and (2) projections from the cluster of cells located dorsal to the brachium conjunctivum that course in the ipsilateral LF. Origins of the pontine reticulospinal tracts arising from the n. reticularis pontis oralis (Poo) have been divided qnto three parts: (1) medial one-third; (2) middle; and (3) ventrolateral. The axons from the medial part descend ipsilaterally via the medial part of the AF, while the axons from the ventrolateral part of the Poo give rise to diffuse descending projections in the AF and LF. The middle part of the Poo has been further subdivided into: (1) dorsal part that gives rise to spinal projections ipsilaterally in the ventrolateral funiculus (VLF); and (2) ventral, particularly its upper part, whose axons descend bilaterally via the DLF. Origins of reticulospinal tracts from the n. reticularis pontis caudalis (Poc) could be divided into three parts: (1) medial; (2) dorsolateral; and (3) ventrolateral. The medial part of the Poc is a source of axons via the medial part of the ipsilateral AF, while the ventrolateral part of the nucleus is a source of axons via the contralateral LF. The spinal projections from the dorsolateral part of the Poc appears to course diffusely in the AF and LF, but with DLF predominance. The n. reticularis gigantocellularis (Gc) was found to be a main medullary source of the spinal projections in the ipsilateral AF, while n. reticularis magnocellularis (Mc) is the major source of the fibers coursing ipsilaterally in the VLF. The most medial part of the Mc descends ipsilaterally via the medial part of the AF, while the ventrolateral part of the nucleus together with the n. reticularis lateralis of Meesen and Olszewski descends ipsilaterally via the DLF. It has also been found that the axons from the n. reticularis paramedianus pass via both the AF and LF with ipsilateral predominance, while the n. reticularis dorsalis and ventralis course via the LF with ipsilateral predominance.     

Funicular organization of avian brainstem-spinal projections.

The purpose of this study was to determine which reticulospinal projections need to be preserved to allow voluntary walking and to differentiate between those pathways descending within the ventrolateral funiculus versus the ventromedial funiculus. Retrogradely transported tracers (True Blue, Fast Blue, Diamidino Yellow dihydrochloride, fluorescein-conjugated dextran-amines) were used alone as discrete funicular injections (4-5 microliters) into the lumbar cord (L1), or in conjunction with a more rostral subtotal lesion of the low thoracic cord, to determine the trajectories of brainstem-spinal projections in adult ducks and geese. No difference was found between the species. The major components of the ventromedial funiculus include projections from the medullary reticular formation, pontine reticular formation, raphe obscurus and pallidus, lateral vestibular nucleus, and interstitial nucleus, and to a minor extent from the locus coeruleus, lateral hypothalamus, and nucleus periventricularis hypothalami. The components of the ventrolateral funiculus (VLF) include projections from the nucleus of the solitary tract, nucleus alatus, pontomedullary reticular formation, raphe pallidus, raphe magnus, locus coeruleus, subcoeruleus, lateral vestibular, and descending vestibular nuclei. The principal descending projections within the dorsolateral funiculus (DLF) arose from the red nucleus, the paraventricular nucleus, locus coeruleus, subcoeruleus, dorsal division of the caudal medullary reticular formation, and raphe magnus. The functional implications of the distribution of these descending pathways are discussed with regard to locomotion. Since birds were able to walk despite bilateral lesion of the DLF or VMF but were unable to walk following a bilateral lesion of the VLF, this suggests that medullary reticulospinal pathways coursing within the VLF are essential for the provision of locomotor drive.     

The origin of reticulospinal fibers in the rat: a HRP study

The distribution as well as morphological characteristics of brain stem reticular neurons projecting to spinal cord both of aminergic and non-aminergic natures in the rat was investigated by means of the retrograde horseradish peroxidase (HRP) method. For the identification of aminergic neurons, a combination of HRP technique and monoamine-oxidase staining as well as a pretreatment of 6-hydroxydopamine and 5,6-dihydroxytryptamine (5,6-DHT) was applied. Following the injection of HRP to the cervical, thoracic and lumbar cord, a remarkable number of neurons in the nuclei reticularis ventralis (rv), reticularis lateralis (RI; Meesen & Olszewski), reticularis gigantocellularis, reticularis pontis caudalis, reticularis pontis oralis (rpo), and a small number of cells in the nucleus reticularis dorsalis dorsalis as well as the mesencephalic reticular nuclei were found to be labeled. In the case of cervical injection, HRP labeled cells were also found in the nucleus reticularis parvocellularis, adjacent to the nucleus tractus spinalis n. trigemini oralis, in which labeled neurons were observed. Within Rl, ventral division of rv and lateral part of rpo, the labeled noradrenaline neurons in A 1, 3 and 7, respectively, were found intermingled with the non-aminergic labeled neurons. Many neurons in the nucleus raphe magnus (ram), obscurus (rao) and pallidus (rap) were labeled. From the fact that there was a marked decrease in the number of the labeled cells in rao, rap, and a slight decrease in ram after 5,6-DHT treatment, it was suggested that the majority of labeled cells in rao, rap and a partial number of labeled cells in ram are serotonergic.     

Brainstem afferents to the oculomotor omnipause neurons in monkey

To determine how saccade-related areas in the brainstem address the saccade generator, we examined the afferents to the nucleus raphe interpositus. This region contains the omnipause neurons, which are pivotal in the generation of saccades. Horseradish peroxidase injected iontophoretically into the nucleus raphe interpositus retrogradely labeled a variety of brainstem nuclei. The greatest numbers of labeled neurons were in the paramedian pontomedullary reticular formation, in the nuclei reticularis gigantocellularis, and paragigantocellularis lateralis. Labeling was more modest but consistent in the interstitial nucleus of Cajal and the adjacent mesencephalic reticular formation, the middle gray of the superior colliculi, the region dorsolateral to the nucleus reticularis tegmenti pontis, and the medial vestibular nucleus. A few neurons were labeled around the habenulopeduncular tract and in the medial portion of the nucleus of the fields of Forel, in the nucleus reticularis medullaris ventralis, and in the spinal nucleus of the trigeminal nerve, the cochlear nucleus, and the superior olivary complex. The distribution and density of labeling suggest that omnipause neurons in the monkey are more intimately connected with other oculomotor structures than those in the cat. In addition, the rhombencephalic reticular afferents to the monkey omnipause neurons are more concentrated in their immediate vicinity than in the cat. The label consistently found dorsolateral to the nucleus reticularis tegmenti pontis may be a newly discovered link in saccade generation.     

Distinguishing rat brainstem reticulospinal nuclei by their neuronal morphology. I. Medullary nuclei

While cytoarchitectonic and hodological investigations suggest that the brainstem reticulospinal nuclei (BRN) are complexly organized, previous Golgi studies claimed that BRN comprise a homogeneous population with respect to neuronal morphology. To determine whether this is indeed the case, neurons of the various BRN of adult albino or hooded rats were either backfilled with horseradish peroxidase (HRP) from spinal injections, stained with a Nissl method or impregnated with a Golgi-Kopsch variant. The results suggest that at least thirteen BRN can be distinguished in the medulla. Some medullary BRN contain neurons whose dendritic arborizations (DA) are radially symmetric (e.g., nucleus reticularis (NR) ventralis pars beta (RVb), NR gigantocellularis (RGc) and nucleus raphe magnus (RaM]. Some BRN contain neurons whose DA exhibit a pronounced dorsomedial to ventrolateral slant (e.g., NR dorsalis (RD) and NR parvocellularis (RPc). The DA of NR paragigantocellularis dorsalis (RPgcd) neurons tend to course dorsally. The DA of nucleus raphe obscurus (RaO) neurons course vertically, while those of NR magnocellularis pars alpha (RMca) and NR magnocellularis pars beta (RMcb) course horizontally. The DA of NR ventralis pars alpha (RVa) may be oriented horizontally also, but sometimes slant from dorsolateral to ventromedial. The DA of NR paramedianus neurons (RPm) are cruciform. The neurons of NR paragigantocellularis lateralis (RPgcl) and of the nucleus raphe pallidus (RaP) exhibit a variety of DA patterns. The neurons of RD, RVa, RMcb and RMca project to the spinal cord with a strong ipsilateral predominance, while those of RVb, RPgcl and RGc project to the spinal cord with a weak ipsilateral predominance. The axons of RPc, RaO, RaP, and RaM neurons exhibit no lateral predominance. RPm neurons project to the cord with a weak contralateral predominance, and RPgcd neurons project to the cord with a strong contralateral predominance. Most medullary BRN project to the spinal cord via the medial longitudinal fasciculus (mlf) and sulcomarginal fasciculus. However, RPgcl, RMca and RMcb also project to the spinal cord via the lateral funiculus. The neurons of RD, RPm and RaM project to the spinal cord exclusively via the lateral or dorsolateral funiculus. Since the various medullary BRN are distinguishable on the basis of neuronal morphology, they may play distinct roles in brainstem modulation of spinal motor, sensory or autonomic activity.     

Lower brainstem afferents to the cat posterior hypothalamus: a double-labeling study

Using a double-immunostaining technique with cholera toxin (CT) as a retrograde tracer, the authors examined the cells of origin and the histochemical nature of lower brainstem afferents to the cat posterior hypothalamus. The posterior hypothalamus, in particular the lateral hypothalamic area, receives substantial afferent projections from: substantia nigra, peripeduncular nucleus, ventral tegmental area, periaqueductal grey, mesencephalic reticular formation, peribrachial region including the locus coeruleus complex, rostral raphe nuclei and the rostral part of the nucleus magnus. In addition, a moderate number of retrogradely labeled neurons was found in: Edinger-Westphal nucleus, nucleus reticularis pontis oralis, nucleus reticularis magnocellularis, caudal lateral bulbar reticular formation around the nucleus ambiguus and lateral reticular nucleus and the nucleus of the solitary tract. The posterior hypothalamus receives: 1) dopaminergic inputs from A8, A9 and A10 cell groups; 2) noradrenergic inputs from A6 and A7 pontine, as well as A1 and A2 bulbar cell groups; 3) adrenergic inputs from C1 cell group in the caudal medulla; 4) serotoninergic inputs from the rostral raphe nuclei (B6, B7 and B8 cell groups); 5) cholinergic inputs from the peribrachial region of the dorsal pontine tegmentum as well as from the nucleus reticularis magnocellularis of the medulla; 6) peptidergic inputs such as methionine-enkephalin, substance P, corticotropin-releasing factor and galanin that originate mainly in the mesencephalic periaqueductal grey, the dorsal raphe nucleus and the peribrachial region of the dorsal pontine tegmentum.     

Nuclei of origin of monoaminergic, peptidergic, and cholinergic afferents to the cat trigeminal motor nucleus: a double-labeling study with cholera-toxin as a retrograde tracer.

The aim of the present study was to determine the brainstem afferents and the location of neurons giving rise to monoaminergic, cholinergic, and peptidergic inputs to the cat trigeminal motor nucleus (TMN). This was done in colchicine treated animals by using a very sensitive double immunostaining technique with unconjugated cholera-toxin B subunit (CT) as a retrograde tracer. After CT injections in the TMN, retrogradely labeled neurons were most frequently seen bilaterally in the nuclei reticularis parvicellularis and dorsalis of the medulla oblongata, the alaminar spinal trigeminal nucleus (magnocellular division), and the adjacent pontine juxtatrigeminal region and in the ipsilateral mesencephalic trigeminal nucleus. We further observed that inputs to the TMN arise from the medial medullary reticular formation (the nuclei retricularis magnocellularis and gigantocellularis), the principal bilateral sensory trigeminal nucleus, and the dorsolateral pontine tegmentum. In addition, the present study demonstrated that the TMN received 1) serotonergic afferents, mainly from the nuclei raphe obscurus, pallidus, and dorsalis; 2) catecholaminergic afferent projections originating exclusively in the dorsolateral pontine tegmentum, including the K?lliker-Fuse, parabrachialis lateralis, and locus subcoeruleus nuclei; further, that 3) methionin-enkephalin-like inputs were located principally in the medial medullary reticular formation (nuclei reticularis magnocellularis and gigantocellularis and nucleus paragigantocellularis lateralis), in the caudal raphe nuclei (Rpa and Rob) and the dorsolateral pontine tegmentum; 4) substance P-like immunoreactive neurons projecting to the TMN were present in the caudal raphe and Edinger-Westphal nuclei; and 5) cholinergic afferents originated in the whole extent of the nuclei reticularis parvicellularis and dorsalis including an area located ventral to the nucleus of the solitary tract at the level of the obex. In the light of these anatomical data, the present report discusses the possible physiological involvement of TMN inputs in the generation of the trigeminal jaw-closer muscular atonia occurring during the periods of paradoxical sleep in the cat.     

Immunohistochemistry and spinal projections of the reticular formation in the northern leopard frog,Rana pipiens

Over 30 nuclei have been identified in the reticular formation of rats, but only a small number of distinct reticular nuclei have been recognized in frogs. We used immunohistochemistry, retrograde tracing, and cell morphology to identify nuclei within the brainstem of Rana pipiens. FluoroGold was injected into the spinal cord, and, in the same frogs, antibodies to enkephalin, substance P, somatostatin, and serotonin were localized in adjacent sections. We identified many previously unrecognized reticular nuclei. The rhombencephalic reticular formation contained reticularis (r.) dorsalis; r. ventralis, pars alpha and pars beta; r. magnocellularis; r. parvocellularis; r. gigantocellularis; r. paragigantocellularis lateralis and dorsalis; r. pontis caudalis, pars alpha and pars beta; nucleus visceralis secundarius; r. pontis oralis, pars medialis and pars lateralis; raphe obscurus; raphe pallidus; raphe magnus; and raphe pontis. The mesencephalic reticular formation contained locus coeruleus-subcoeruleus, r. cuneiformis, r. subcuneiformis, raphe dorsalis-raphe centralis superior, and raphe linearis. Thus, the reticular formation of frog, which is an anamniote, is organized complexly and is similar to the reticular formation in amniotes. Because many of these nuclei may be homologous to reticular nuclei in mammals, we used mammalian terminology for frog reticular nuclei. J. Comp. Neurol. 404:387–407, 1999. © 1999 Wiley-Liss, Inc.     

Nucleus cuneiformis and pain modulation: anatomy and behavioral pharmacology

The anatomical substrate and behavioral pharmacology of stimulation-produced analgesia resulting from electrical stimulation of the pontomesencephalic nucleus cuneiformis (NCF) was determined in the present study. Maximum increase in nociceptive tail-flick latencies following NCF stimulation occurred during the first 5 min post stimulation and decreased afterwards. The increased reflex latency could be attenuated by prior treatment with the narcotic antagonist, naloxone or the cholinergic antagonist, scopolamine. The anatomical projections of NCF were identified in autoradiographic and histochemical studies. Ipsilateral fibers coursed caudal from the NCF injection site through the ventral pontine reticular formation to innervate nucleus raphe magnus and the ipsilateral nucleus magnocellularis. At rostral medullary levels fibers coursed dorsolateral to innervate the ipsilateral nucleus reticularis parvocellularis. Descending contralateral fibers crossed through the decussation of the superior cerebellar peduncle, then coursed ventrolaterally projecting to the contralateral nucleus magnocellularis. Two primary groups of ascending fibers were observed. The dorsally located group ascended through the central tegmental tract projecting to the dorsal raphe, ipsilateral periaqueductal gray, nucleus parafascicularis and centromedianus, the intermediolateral and lateral thalamic nuclei. The ventral group coursed ventrolateral from the injection site projecting to the substantia nigra, zona compacta, ventral tegmental area of Tsai, zona incerta, Fields of Forel, lateral hypothalamic nucleus and nucleus reuniens. These anatomic and behavioral data suggest that NCF plays an important role in sensory/motor integration relevant to pain transmission.     

Organization of tyrosine hydroxylase- and serotonin-immunoreactive brainstem neurons with axon collaterals to the periaqueductal gray and the spinal cord in the rat

Retrograde tracing and immunocytochemistry were used to examine the axon collateralization of brainstem serotonin (5-HT) and norepinephrine (NE) cells to the periaqueductal gray (PAG) and spinal cord. Tyrosine hydroxylase (TH)-immunofluorescent neurons which collateralize to the PAG and the cervical spinal cord were found in all brainstem catecholamine cell groups previously shown to contain neurons which project to the spinal cord, including the A5 and A7 cell groups, locus coeruleus, subcoeruleus and the C1 cell group. Many TH-immunofluorescent cells which project to the PAG but not to the spinal cord were also found. The region of the nucleus raphe magnus (NRM) also contained many neurons retrogradely labeled from the PAG. These overlapped with the distribution of spinally projecting 5-HT-immunofluorescent cells in the NRM, however, less than 1% of the PAG projecting cells in this region were 5-HT-immunofluorescent. In contrast, many 5-HT-immunofluorescent cells in the more rostral nucleus raphe pontis and nucleus raphe dorsalis were retrogradely labeled from the PAG but not from the spinal cord. Finally, a population of neurons in the NRM and adjacent reticular formation and in the region of several pontomedullary catecholamine cell groups collateralized to the PAG and spinal cord, but were neither 5-HT nor TH-immunofluorescent. Taken together, these findings raise the possibility that the noradrenergic contribution to the spinal antinociceptive effects produced by PAG electrical stimulation results, in part, from antidromic activation of brainstem noradrenergic neurons that have axon collaterals projecting to the PAG and spinal cord. In contrast, the 5-HT contribution to the spinal antinociceptive effects produced by PAG electrical stimulation is more likely to derive, as previously proposed, from orthodromic activation of raphe-spinal serotonergic axons.     

Somatodendritic and axonal anatomy of intracellularly labeled serotonergic neurons in the rat medulla

A knowledge of the anatomy of medullary serotonergic cells is critical to understanding local and brainstem circuits in which these cells participate. Serotonergic neurons (n = 16) were identified, as previously described (Mason [1997] J. Neurophysiol. 77:1087–1098) by their slow and steady background discharge in halothane anesthetized rats. Neurons were then intracellularly labeled with Neurobiotin and visualized with 3,3'diaminobenzidine. The validity of the physiological identification of serotonergic cells was confirmed by processing two neurons that were physiologically characterized as serotonergic for serotonin immunoreactivity; both tested cells contained immunoreactive serotonin. The dendrites and axon of each labeled cell were reconstructed by using a three-dimensional computerized system. Somata were small or medium in size and had fusiform, triangular, or multipolar shapes. The dendritic arbor was constricted with most dendrites extending for less than 500 μm from the soma. All labeled axons projected caudally and travelled in the ventrolateral medulla, either dorsal or ventral to the lateral reticular nucleus. Most cells had collaterals and/or dense axonal swellings in the nucleus reticularis gigantocellularis, nucleus reticularis magnocellularis, raphe magnus, and the ventrolateral medulla. Non-local collaterals and swellings were also observed in the nucleus reticularis gigantocellularis and in the ventrolateral medulla at all medullary levels. The results demonstrate that 1) the dendrites of serotonergic cells are restricted to raphe magnus and the ventral part of nucleus reticularis magnocellularis; and 2) serotonergic cells project to medullary nuclei that contain bulbospinal cells which project to dorsal, intermediate, and ventral horns. Serotonergic cell projections to brainstem sites may mediate the integration of sensory, autonomic, and motor modulation at the brainstem level. J. Comp. Neurol. 389:309–328, 1997. © 1997 Wiley-Liss, Inc.     

Afferent connections of the rostral medulla of the cat: A neural substrate for midbrain-medullary interactions in the modulation of pain

In order to study the organization of the rostral medulla of the cat and its contribution to pain control mechanisms, we have examined the afferent connections of the midline nucleus raphe magnus (NRM), the laterally located nucleus reticularis magnocellularis (Rmc), and the nucleus reticularis gigantocellularis (Rgc) located dorsal to Rmc. Iontophoretic injections of HRP were made into the three regions; the distribution of retrogradely labeled neurons in brainstem and spinal cord was then mapped. While significant differences characterize the source of afferents to Rgc and NRM/Rmc, there is little to distinguish that between NRM and Rmc. The predominant spinal projection is to Rgc; fewer labeled neurons were recorded after injections into Rmc. In contrast, no significant direct spinal projection to NRM was found. All three regions receive input from widespread areas within the medullary and pontine reticular formation. The most pronounced differences in the distribution of retrogradely labeled neurons were found in the midbrain. The major projection to both NRM and Rmc derives from the periaqueductal gray (PAG) and from the adjacent nucleus cuneiformis. Labeled cells are concentrated in the dorsal and lateral PAG; few are found in the ventrolateral PAG. In contrast, Rgc receives few afferents from the PAG; however, after Rgc injections, many cells were recorded in the deep layers of the contralateral tectum. None of the injection sites produced significant labeling of the catecholamine-rich dorsolateral pontine tegmentum or of the nucleus raphe dorsalis. The demonstration of significant PAG projections to NRM/Rmc provides anatomical evidence for the hypothesis that opiate and stimulation-produced analgesia involves connections from PAG to neurons of NRM and Rmc which, in turn, inhibit spinal nociceptors.     

Evidence for leucine-enkephalin immunoreactive neurons in the medulla which project to spinal cord in squirrel monkey

Rhodamine-labeled latex microsphere injections were combined with horserasish peroxidase immunohistochemistry in squirrel monkeys to reveal neurons in the medullary raphe and medial reticular formation which projected to spinal cord and were positive for leucine-enkephalin-like immunoreactivity. Double-labeled cells were found primarily in nucleus raphe magnus, the reticular nucleus magnocellularis, and the lateral reticular nucleus. These results provide evidence for a descending projection from enkephalin-containing cells of the rostral ventral medulla, a region which has been strongly implicated in antinociception.     

Arborization of single axons of the spinal dorsolateral funiculus to the contralateral superficial dorsal horn

This study provides an anatomical basis for the observation that a unilateral lesion of the spinal dorsolateral funiculus (DLF) can reduce the inhibitory effect of electrical stimulation of the nucleus raphe magnus (NRM) on dorsal horn nociceptive neurons located caudal and contralateral to the lesion. We injected the anterograde tracer Phaseolus vulgaris leucoagglutinin (PHA-L) into the NRM and traced the arborization of single DLF axons in the spinal gray matter. Although the majority of DLF axons arborized in the dorsal horn ipsilaterally, we found some axons which entered the spinal gray matter, traversed the gray matter to the central canal and then abruptly changed direction and coursed to the contralateral superficial dorsal horn. Very few branches were given off en route. These data indicate that some raphe-spinal axons may selectively influence the firing of neurons of the superficial dorsal horn, contralateral to the DLF in which they descend the spinal cord.     

Projections of the dorsal raphe nucleus to the brainstem: PHA-L analysis in the rat

Early studies that used older tracing techniques reported exceedingly few projections from the dorsal raphe nucleus (DR) to the brainstem. The present report examined DR projections to the brainstem by use of the anterograde anatomical tracer Phaseolus vulgaris leucoagglutinin (PHA-L). DR fibers were found to terminate relatively substantially in several structures of the midbrain, pons, and medulla. The following pontine and midbrain nuclei receive moderate to dense projections from the DR: pontomesencephalic central gray, mesencephalic reticular formation, pedunculopontine tegmental nucleus, medial and lateral parabrachial nuclei, nucleus pontis oralis, nucleus pontis caudalis, locus coeruleus, laterodorsal tegmental nucleus, and raphe nuclei, including the central linear nucleus, median raphe nucleus, and raphe pontis. The following nuclei of the medulla receive moderately dense projections from the DR: nucleus gigantocellularis, nucleus raphe magnus, nucleus raphe obscurus, facial nucleus, nucleus gigantocellularis-pars alpha, and the rostral ventrolateral medullary area. DR fibers project lightly to nucleus cuneiformis, nucleus prepositus hypoglossi, nucleus paragigantocellularis, nucleus reticularis ventralis, and hypoglossal nucleus. Some differences were observed in projections from rostral and caudal parts of the DR. The major difference was that fibers from the rostral DR distribute more widely and heavily than do those from the caudal DR to structures of the medulla, including raphe magnus and obscurus, nucleus gigantocellularis-pars alpha, nucleus paragigantocellularis, facial nucleus, and the rostral ventrolateral medullary area. A role for the dorsal raphe nucleus in several brainstem controlled functions is discussed, including REM sleep and its events, nociception, and sensory motor control. © Wiley-Liss, Inc.     

Identification and somatotopic organization of nuclei projecting via the dorsolateral funiculus in rats: A retrograde tracing study using HRP slow-release gels

Horseradish peroxidase (HRP) in slow-release gel was unilaterally implanted in the transected dorsolateral funiculus (DLF) of either cervical, midthoracic, lumbar or sacral spinal cord levels of adult male rats. Cell mappings were made of all brain areas projecting through the DLF. Following cervical implants, dense labelling was observed within a band along the dorsolateral border of the inferior olive, locus coeruleus, the paralemniscal reticular formation, the mesencephalic central gray, the red nucleus and the paraventricular nucleus of the hypothalamus. The DLF projection from the central gray consisted of a rostrocaudal line of cells extending from the level of the mesen-/diencephalic junction to the rostral red nucleus. The somatotopic organization of labelled nuclei was assessed by comparing the pattern of filled cells following HRP implants at various cord levels. The potential role of these areas in pain modulation was discussed in light of their descending projections through the DLF of the spinal cord.