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.     

Brainstem origin of serotonin- and enkephalin-immunoreactive afferents to the opossum's cerebellum

Previous studies have described the distribution of serotonin- and enkephalin-immunoreactive elements in the posterior lobe vermis of the opossum's cerebellum. In the present study we have used a double labeling paradigm which combines the retrograde transport of horseradish peroxidase (HRP) with serotonin and enkephalin immunohistochemistry to determine the brainstem origin of serotoninergic and enkephalinergic neurons that project to the opossum's cerebellar cortex. Subsequent to HRP injections into the posterior lobe vermis, widespread areas of the medulla and pons were found to contain retrogradely labeled neurons. Serotonin-immunoreactive somata are present primarily in the raphe nuclei and the adjacent reticular formation. Enkephalinergic neurons were numerous in the raphe nuclei, medial accessory olive, gigantocellular reticular formation, locus coeruleus, and the nucleus of the trapezoid body. However, serotoninergic neurons that project to the cerebellum were located only in the medullary pyramids and the reticular formation adjacent to the raphe. Double-labeled enkephalinergic neurons were located 1) within the medullary pyramids, 2) throughout the extent of the caudal medial accessory olive, 3) in the rostral subnucleus a of the medial accessory olive, 4) in the nucleus reticularis gigantocellularis pars ventralis, 5) in the nucleus reticularis lateralis, and 6) in the nucleus reticularis ventralis lateral to the inferior olivary complex. These results indicate that although neurons containing serotonin and enkephalin immunoreactivity may be present in some of the same pontine and medullary nuclei, those serotoninergic and enkephalinergic neurons that project to the cerebellum are present primarily in restricted and spatially separate regions of the caudal medulla.     

Projections from the brain stem reticular formation to laminae I and II of the spinal cord. Studies using light and electron microscopic techniques in the North American opposum

The horseradish peroxidase and autoradiographic methods show that laminae I and outer II are innervated by the nucleus reticularis gigantocellularis pars ventralis, and the nucleus reticularis pontis pars ventralis. Both areas contain neurons of the indolamine type and probably account for the indolamine-like varicosities which are present within laminae I and II. Degeneration material prepared for electron microscopy showed that axons from the above nuclei end on small dendritic shafts and spines as well as on vesicle-filled profiles. The terminals identified formed asymmetrical contacts and contained clear as well as dense-cored vesicles. No terminals were present within glomuruli. A projection to laminae I and outer II also arises within the dorsolateral pons and several lines of evidence suggest that it is catecholaminergic. The electron microscope revealed that axons from the dorsolateral pons are fairly numerous within laminae I and II, but that terminal contacts are relatively rare. Those present are asymmetrical and alternate with intermediate-sized dendrites. They contain clumps of clear, spherical vesicles as well as larger vesicles with a variety of dense cores.     

The origin of descending pathways in the dorsolateral funiculus of the spinal cord of the cat and rat: further studies on the anatomy of pain modulation.

There is considerable evidence that the dorsolateral funiculus (DLF) of the spinal cord contains descending pathways critical for both opiate and brainstem stimulation-produced analgesia. To obtain a comprehensive map of brainstem neurons projecting to the spinal cord via the DLF, large injections of horseradish peroxidase (HRP) were made into the lumbosacral spinal cord of cat and rat. These injections were made caudal to midthoracic lesions which spared only a single DLF or ventral quadrant (VQ); thus only those neurons whose axons descended in the spared funiculus would be labelled. Cells with descending axons in the VQ were concentrated in the medullary nucleus raphe pallidus and obscurus, nucleus retroambiguus and in various subregions of the reticular formation including the nucleus reticularis ventralis, gigantocellularis, magnocellularis, pontis caudalis and pontis oralis. Significant numbers of neurons were also found in medial and lateral vestibular nuclei and in several presumed catecholamine-containing neurons of the dorsolateral pons. In the rat, but not in the cat, considerable numbers of cells are present in the mesencephalic reticular formation just lateral to the periaqueductal gray. In both species, some cells were found in the paraventricular nucleus of the hypothalamus. Brainstem cells projecting in the DLF were concentrated in the nucleus raphe magnus and in the adjacent nucleus reticularis magnocellularis, ipsilateral to the spared funiculus. Significant numbers of cells were found in the dorsolateral pons, differing somewhat in their distribution from those projecting in the VQ. DLF-projecting cells were also present in the ipsilateral Edinger-Westphal nucleus and periaqueductal grey contralateral red nucleus of the midbrain and in the ipsilateral hypothalamus. Smaller projections from other sites are described. These results are discussed in terms of the differential contribution of several brainstem neuronal groups, including the serotonergic nucleus, raphe magnus, the ventromedial reticular formation of the medulla, and various catecholamine-containing neurons of the dorsolateral pontine tegmentum to the analgesia produced by opiates and electrical brain stimulation.     

The organization of monoamine neurons within the brainstem of the north american opossum (didelphis virginiana)

The distribution of monoamine-containing neurons within the brain of the opossum is described using the Falck-Hillarp histofluorescence technique. Catecholamine-containing neurons are organized into four groups. The medulla contains one group which is located dorsolateral to the lateral reticular nucleus and ventrolateral to the dorsal vagal nucleus. The second collection is found within the pons and includes both the locus coeruleus and a region continuous with it referred to as the nucleus coeruleus, pars α. The third aggregate includes the substantia nigra, the ventral tegmental area, and the mesencephalic reticular formation and a fourth group is located within the periventricular and dorsal paraventricular nuclei of the hypothalamus. The indoleamine-containing cell bodies are distributed within the nuclei raphe obscurus, pallidus, magnus, dorsalis, and the nuclei linearis and superior centralis except at certain pontine levels where they appear laterally within the reticular formation. A number of small intensely fluorescent (SIF) cells are present within the connective tissue surrounding the brain and its blood vessels as well. Although certain differences are present, the didtribution of monoamine neurons in the American opossum conforms generally to that described for the placental mammals studied to date.     

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.     

Brainstem projections to the facial nucleus of the opossum. A study using axonal transport techniques

The horseradish peroxidase and autoradiographic techniques have been used to determine the origin and intranuclear termination of brainstem axons projecting to the facial nucleus of the opossum and to define networks which could be utilized in some oral-facial behaviors. Two regions of the midbrain have dense projections to the facial nucleus. One region is the ventral periaqueductal gray and adjacent interstitial nucleus of the medial longitudinal fasciculus which project bilaterally to those areas of the facial nucleus supplying auricular and cervical musculature. A second is the paralemniscal zone of the caudolateral midbrain which innervates the same areas of the contralateral facial nucleus. The red nucleus and/or the adjacent tegmentum send a less dense projection to those regions of the contralateral facial nucleus which innervate buccolabial and zygomatic muscles. The dorsolateral pons (the parabrachial complex, the nucleus locus coeruleus, pars alpha, and the nucleus sensorius n. trigemini, pars dorsalis) projects densely to those areas of the ipsilateral facial nucleus which innervate buccolabial and zygomatic musculature. In contrast, the nucleus reticularis pontis, pars ventralis, projects bilaterally to parts of the facial nucleus supplying auricular and cervical muscles. There was evidence of some rostral to caudal organization in the latter projection. Neurons in medial parts of the lateral reticular formation project bilaterally to the facial nucleus. Those within the nucleus reticularis parvocellularis and the rostral nucleus reticularis medullae oblongatae ventralis innervate areas supplying buccolabial and zygomatic muscles. Neurons in the nucleus reticularis medullae oblongatae ventralis located caudal to the obex favor regions of the facial nuclei which supply auricular and cervical muscles. Neurons in the nucleus reticularis medullae oblongatae dorsalis and lamina V of the medullary and spinal dorsal horns project ipsilaterally to the facial nucleus in a manner suggesting that information from specific cutaneous areas reaches neurons supplying the muscles deep to them. The brainstem-facial connections are discussed in relation to the functionally diverse roles served by the facial nucleus in oral-facial behavior.     

Spinal projections from the medullary reticular formation of the North American opossum: heterogeneity

Retrograde and orthograde transport techniques show that the nucleus reticularis gigantocellularis pars ventralis and the nucleus reticularis gigantocellularis project the entire length of the spinal cord. Double-labelling methods show that some of the neurons in each area innervate both cervical and lumbar levels. There is evidence, however, that neurons in the lateral part of the nucleus reticularis gigantocellularis pars ventralis and the dorsal extreme of the nucleus reticularis gigantocellularis project mainly to cervical and thoracic levels. The autoradiographic method shows that the above nuclei supply direct innervation to somatic and autonomic motor columns as well as to laminae V-VIII and X. The nucleus reticularis gigantocellularis pars ventralis provides additional projections to lamina I and the outer part of lamina II. Several areas of the medullary reticular formation project mainly, and in some cases exclusively, to cervical and thoracic levels. These areas include the nucleus reticularis parvocellularis, the nucleus reticularis lateralis, the nucleus retrofacialis, the nucleus ambiguus, the nucleus lateralis reticularis, caudal parts of the nuclei reticularis medullae oblongatae dorsalis and ventralis, and the nucleus supraspinalis. Autoradiographic experiments reveal that neurons in the ventrolateral medulla, particularly rostrally (the nucleus reticularis lateralis and neurons related to the nucleus lateralis reticularis), innervate sympathetic nuclei. Our results indicate that spinal projections from bulbar areas of the reticular formation are more complicated than previously supposed. Axons from separate areas project to different spinal levels and in some cases to different nuclear targets. These data are in conformity with the evolving concept of reticular heterogeneity.     

Origins and terminations of bulbospinal axons that contain serotonin and either enkephalin or substance-P in the North American opossum.

We have shown previously that some enkephalin, substance-P, and serotoninergic neurons in the medullary raphe and adjacent reticular formation project to the spinal cord in the opossum. In the present study we have combined the retrograde transport of True Blue and immunofluorescence histochemistry to determine whether methionine enkephalin or substance-P containing bulbospinal neurons are serotoninergic. Furthermore, we have used the same immunofluorescence protocol to determine whether spinal axons contain the same substances. Neurons that immunostained for both enkephalin and serotonin were observed in many brainstem nuclei. However, those that projected to the spinal cord were limited to the nuclei raphe magnus and obscurus, and the ventral part of nucleus reticularis gigantocellularis, pars ventralis. Neurons that immunostained for both substance P and serotonin were fewer in number, but some of the ones in the above nuclei and within the nucleus raphe pallidus, projected to the spinal cord. Spinal axons exhibiting both enkephalin- and serotonin-like immunoreactivity were observed in the superficial laminae of the dorsal horn, lamina X, and the intermediolateral cell column, whereas those showing both substance-P and serotonin-like immunoreactivity were seen primarily in lamina X, the intermediolateral cell column, and the ventral horn. Some of the axons in the ventral horn were in close apposition to presumed motoneurons. Comparison of the above results with those obtained from previous studies of bulbospinal projections has allowed us to infer the origins of axons that innervate different spinal targets.     

Spinal projections from the mesencephalic and pontine reticular formation in the North American Opossum: a study using axonal transport techniques.

The results from several experimental approaches lead to the following conclusions. The nucleus cuneiformis projects to at least lumbar levels of the spinal cord. Its axons course through the ipsilateral sulcomarginal and ventral funiculi to distribute within lamina VIII and adjacent portions of lamina VII. Neurons within the nucleus reticularis pontis (RP), particularly within more medial parts of the nucleus, project through comparable routes to the same laminae. In addition, however, neurons within the lateral and dorsolateral RP relay through the lateral and dorsolateral funiculi, ipsilaterally, and the dorsolateral funiculus, contralaterally. Axons could be traced from the dorsolateral tracts to laminae IV through VII, lamina X and, in some instances, to laminae I and II. Injections of the dorsolateral pons also label the intermediolateral cell column and an area presumed to be the sacral parasympathetic nucleus. Many of the neurons which contribute to the contralateral bundle are located adjacent to the ventral nucleus of the lateral lemniscus. The nucleus reticularis gigantocellularis projects mainly via the sulcomarginal, ventral and lateral funiculi to laminae VIII and adjacent portions of lamina VII. The nucleus reticularis gigantocellularis pars ventralis innervates the same laminae; but, in addition, projects heavily to laminae I and II, to lateral portions of laminae IV through VII; to laminae IX and X and to the intermediolateral cell column. Axons destined for laminae I and II, as well as IV through VII and X, traverse the dorsolateral funiculi as described for the cat by Basbaum et al. ('78). Neurons within the nucleus reticularis parvocellularis project to cervical levels, mainly through the ventral funiculi. In general our results show that reticulospinal projections are more complex than suggested by degeneration methods and that laminae I, II. lateral parts of laminae IV-VII, laminae IX and X, as well as the intermediolateral cell column and sacral parasympathetic nucleus are targets of axons from specific areas.     

Serotonergic innervation of the forebrain in the North American opossum

The forebrain distribution of axons showing serotonin-like immunoreactivity was studied in the North American opossum. Serotonergic innervation of the hypothalamus was extensive, particularly within the ventromedial nucleus, the periventricular nucleus and the rostral supraoptic nucleus. Serotonergic axons were also present within the fields of Forel and zona incerta, but they tended to avoid parts of the subthalamic nucleus. In the thalamus serotonergic innervation was dense within the midline nuclei (e.g. the central, intermediate dorsal and rhomboid nuclei) and the ventral lateral geniculate nucleus, but relatively sparse in some of the nuclei more readily associated with specific functions (e.g. the ventrobasal nucleus). Serotonergic axons innervate most areas of the rostral and dorsal forebrain. Areas containing the heaviest innervation included the interstitial nucleus of the stria terminalis and the lateral septal nucleus. Serotonergic innervation of the neocortex varied markedly from region to region and within different layers of the same regions. The retrograde transport of True Blue combined with immunofluorescence for localization of serotonin revealed that serotonergic axons within the forebrain arise mainly within the dorsal raphe and superior central nuclei, but that some originate within the midbrain and pontine reticular formation and the locus coeruleus, pars alpha. Neurons of the raphe magnus and obscurus also innervate the forebrain, but few of them are serotonergic. The use of horseradish peroxidase as a retrograde marker provided evidence that raphe projections to the forebrain are topographically organized. Our results suggest that serotonergic projections to the forebrain, like those to the spinal cord, are connectionally heterogeneous.     

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.     

Spinal projections from the lower brain stem in the cat as demonstrated by the horseradish peroxidase technique. II. projections from the dorsolateral pontine tegmentum and raphe nuclei

The descending projections to the spinal cord arising from the dorsolateral pontine tegmentum and brain stem raphe nuclei have been investigated by means of the horseradish peroxidase (HRP) technique. Particular attention was taken to clarify the cells of origin and the funicular trajectory of these spinal projections.After injections of HRP into the spinal cord, a significant number of HRP labeled neurons were observed in the following dorsolateral pontine tegmental structures: (1) an area ventral to the nucleous cuneiformis; (2) principal locus coeruleus; (3) locus coeruleus α; (4) locus subcoeruleus; (5) Kölliker-Fuse nucleus; and (6) nucleus parabrachialis lateralis. As a rule, the projections are ipsilateral and the descending fibers course in the ventral part of the lateral funiculus.As concerns the raphe-spinal projections, we have demonstrated that the nucleus raphe dorsalis also sends axons to the cervical segment of the spinal cord. Furthermore, in accord with previous reports, HRP labeled cells were also identified in the nucleus raphe magnus, pallidus and obscurus, but not in the nucleus raphe centralis superior and pontis.On the whole the present study further clarified the organization of spinal projections from the dorsolateral pons and raphe nuclei and provided some additional anatomical data for the physiology of the tegmentospinal and raphe-spinal projections.     

微量海人酸注入大鼠延髓网状背侧亚核诱导的Fos表达及其与NADPH-d的共存

目的：研究大鼠网状背侧亚核的功能传出通路及该通路上NADPH-d的分布。方法：将微量海人酸注入大鼠延髓网状背侧亚核，2h后灌注，脑片进行NADPH-d组织化学和Fos免疫组化染色。结果：Fos阳性细胞主要分布于中缝背核(DR)、中缝大核(NRM)、蓝斑(LC)、中脑导水管周围灰质腹外侧(vlPAG)和下丘脑室旁核。Fos／NADPH-d双标细胞主要分布于巨细胞网状核、下丘脑室旁核和中缝大核。结论：大鼠延髓网状背侧亚核与脊髓上中枢存在广泛的功能联系，而且在延髓网状背侧亚核的功能传出通路上有一氧化氮合酶的分布。     

Collateralization of descending pathways from the brainstem to the spinal cord in a lizard, Varanus exanthematicus

With the multiple fluorescent retrograde tracer technique, the collateralization in the spinal cord of descending supraspinal pathways was studied in a lizard, Varanus exanthematicus. Fast Blue (FB) gels were implanted unilaterally in the spinal gray matter of the cervical enlargement and Nuclear Yellow (NY) gels were implanted ipsilaterally in two series of experiments in all spinal funiculi of the lumbar enlargement or in midthoracic spinal segments, respectively. All brainstem nuclei known to project to the spinal cord in reptiles were found to give rise to branching axons that may influence widely separate levels of the spinal cord. The number of double-labeled FB-NY cells varied in these brainstem nuclei from none to half the number of neurons projecting to the cervical enlargement. Highly collateralizing projections (expressed as the percentage of double-labeled neurons, DL) were found to arise from the nucleus raphes inferior, the contralateral nucleus reticularis superior pars lateralis, the contralateral nuclei vestibulares ventromedialis and descendens, and the ipsilateral nucleus reticularis inferior pars ventralis. A lower percentage of DL neurons was noted for the contralateral nucleus ruber and bilaterally for the nucleus reticularis medius and nucleus reticularis inferior. Extensive brainstem projections directed to cervical and high thoracic spinal levels originate from the area lateralis hypothalami, the nucleus of the fasciculus longitudinalis medialis, the contralateral nucleus cerebellaris medialis, and from the nucleus tractus solitarii. Projections preferentially directed to midthoracic or lower levels of the spinal cord were found to arise from the ipsilateral locus coeruleus, the contralateral nucleus reticularis superior pars lateralis, the nucleus reticularis inferior pars ventralis, the nucleus reticularis inferior, and the nucleus raphes inferior. In contrast to findings in mammals, in Varanus exanthematicus the red nucleus, the nucleus vestibularis ventrolateralis, and certain parts of the reticular formation did not display a clear-cut somatotopic organization. In general two different patterns of collateralization can grossly be discerned: a gradual decrease of spinal collaterals caudalward, which can be interpreted as a certain specificity of such projections; and a constant number of collateral nerve fibers throughout the spinal cord that can be interpreted as either a nonspecific or, in contrast, a highly specific system, focussed exclusively on the cervical and lumbar enlargements.     

Afferents to brain stem nuclei (brain stem raphe, nucleus reticularis pontis caudalis and nucleus gigantocellularis) in the rat as demonstrated by microiontophoretically applied horseradish peroxidase

Using a retrograde tracer technique with microiontophoretically applied horseradish peroxidase (HRP), afferent projections to the brain stem raphe nuclei (BR, raphe magnus, pallidus and obscurus) and to two adjacent reticular nuclei, nucleus reticularis pontis caudalis (nRPC) and nucleus gigantocellularis (nGC) were identified. The most striking difference between the afferent projections to the BR and the adjacent nuclei as determined by this method is that afferents to the BR originate primarily from structures rostral to the pons, especially the mesencephalic central gray and the dorsal and ventral tegmentum. In contrast, the two reticular nuclei studied (nGC and nRPC) received afferent projections within or caudal to the pons-medulla. For example, the nGC receives prominent afferent projections from the gray matter of the spinal cord. In addition, evidence for interconnections between all of the adjacent nuclei (BR, nGC and nRPC) was found. Such afferent projections are compatible with the notion that the brain stem raphe nuclei may serve as connections within the brain stem for a descending system, while the nGC may be a relay in a feedback loop between the spinal cord and the reticular formation.     

Subcortical projections to the centromedian and parafascicular thalamic nuclei in the cat

The primary objective of this study is to identify the totality of input to the centromedian and parafascicular (CM-Pf) thalamic nuclear complex. The subcortical projections upon the CM-Pf complex were studied in the cat with three different retrograde tracers. The tracers used were unconjugated horseradish peroxidase (HRP), horseradish peroxidase conjugated to wheat germ agglutinin (WGA-HRP), and rhodamine-labeled fluorescent latex microspheres (RFM). Numerous subcortical structures or substructures contained labeled neurons with all three tracing techniques. These labeled structures included the central nucleus of the amygdala; the entopeduncular nucleus; the globus pallidus; the reticular and ventral lateral geniculate nuclei of the thalamus; parts of the hypothalamus including the dorsal, lateral, and posterior hypothalamic areas and the ventromedial and parvicellular nuclei; the zona incerta and fields of Forel; parts of the substantia nigra including the pars reticularis and pars lateralis, and the retrorubral area; the pretectum; the intermediate and deep layers of the superior colliculus; the periaqueductal gray; the dorsal nucleus of the raphe; portions of the reticular formation, including the mesencephalic, pontis oralis, pontis caudalis, gigantocellularis, ventralis, and lateralis reticular nuclei; the nucleus cuneiformis; the marginal nucleus of the brachium conjunctivum; the locus coeruleus; portions of the trigeminal complex, including the principal sensory and spinal nuclei; portions of the vestibular complex, including the lateral division of the superior nucleus and the medial nucleus; deep cerebellar nuclei, including the medial and lateral cerebellar nuclei; and lamina VII of the cervical spinal cord. Moreover, the WGA-HRP and rhodamine methods (known to be more sensitive than the HRP method) revealed several afferent sources not shown by HRP: the anterior hypothalamic area, ventral tegmental area, lateral division of the superior vestibular nucleus, nucleus interpositus, and the nucleus praepositus hypoglossi. Also, the rhodamine method revealed labeled neurons in laminae V and VI of the cervical spinal cord.     

Brainstem reticular nuclei that project to the cerebellum in rats: a retrograde tracer study.

The nuclear origins of projections from the brainstem reticular formation to the cerebellum were examined using four retrograde tracer substances: horseradish peroxidase, wheat germ agglutinin-horseradish peroxidase conjugate, Fluoro-Gold, and rhodamine beads. Tracer injections were made into each of the three major longitudinal zones of the cerebellar cortex (vermis, paravermal hemisphere, and lateral hemisphere) as well as into the various deep cerebellar nuclei. Counts of retrogradely labeled cells were done on a large sample of select cases. The data generated by these cell counts indicate that the strongest reticulocerebellar projections arise from the three specialized pre-cerebellar reticular nuclei: the lateral reticular nucleus, the medullary paramedian reticular nucleus, and the reticulotegmental nucleus. The presumed noradrenergic locus coeruleus (A6 cell group) was also densely packed with retrogradely labeled neurons. However, strong reticulocerebellar projections also arose from other presumed catecholamine cell groups such as those in the ventrolateral medulla (the A1/C1 complex) and the caudal pons (A5). Substantial cerebellar projections originated from most of the various presumed serotonergic brainstem raphe cell groups (particularly raphe obscurus in the medulla), as well as from the presumed cholinergic Ch5 cell group (the pedunculopontine pars compactus nucleus). Labeled cells were also seen in several nonaminergic isodendritic reticular nuclei thought to be involved in visuomotor activity (e.g. paragigantocellularis dorsalis, raphe interpositus, and the pontine dorsomedial tegmental area), as well as in the lateral reticular zone of the medulla and lower pons (reticularis dorsalis and parvocellularis). Tracer injections into the deep nuclei produced relatively greater numbers of labeled neurons in large-celled medial reticular nuclei associated with skeletomotor activity, such as gigantocellularis, magnocellularis, and pontis caudalis. Reticular nuclei conspicuous in their lack of projections to the cerebellum included reticularis ventralis in the medulla, pontis oralis, and both subdivisions of the midbrain reticular formation (cuneiformis and subcuneiformis). As a whole, the various isodendritic reticular nuclei project most strongly to midline cerebellar structures (vermal cortex or fastigial nuclei), less strongly to the paravermal cortex or interposed nuclei, and least strongly to the lateral cortex or dentate nucleus. Within individual reticular nuclei, the morphology of labeled neurons is identical to that reported previously by this laboratory subsequent to spinal or cortical HRP injections, thus strengthening this laboratory's hypothesis that the various brainstem reticular nuclei can be distinguished on the basis of neuronal morphology.     

Brainstem neurons with descending projections to the spinal cord of two elasmobranch fishes: Thornback guitarfish,Platyrhinoidis triseriata,and horn shark,Heterodontus francisci

We studied two cartilaginous fishes and described their brainstem supraspinal projections because most nuclei in the reticular formation can be identified that way. A retrogradely transported tracer, horseradish peroxidase or Fluoro-Gold, was injected into the spinal cord of Platyrhinoidis triseriata (thornback guitarfish) or Heterodontus fransisci (horn shark). We described labeled reticular cells by their position, morpohology, somatic orientation, dendritic processes, and laterality of spinal projections. Nineteen reticular nuclei have spinal projections: reticularis (r.) dorsalis, r. ventralis pars α and β, r. gigantocellularis, r. magnocellularis, r. parvocellularis, r. paragigantocellularis lateralis and dorsalis, r. pontis caudalis pars α and β, r. pontis oralis pars medialis and lateralis, r. subcuneiformis, r. peduncularis pars compacta, r. subcoeruleus pars α, raphe obscurus, raphe pallidus, raphe magnus, and locus coeruleus. Twenty nonreticular nuclei have spinal projections: descending trigeminal, retroambiguus, solitarius, posterior octaval, descending octaval, magnocellular octaval, ruber, Edinger-Westphal, nucleus of the medial longitudinal fasciculus, interstitial nucleus of Cajal, latral mesencephalic complex, periventricularis pretectalis pars dorsalis, central pretectal, ventromedial thalamic, posterior central thalamic, posterior dorsal thalamic, the posterior tuberculum, and nuclei B, F, and J. The large number of distinct reticular nuclei with spinal projections corroborates the hypothesis that the reticular formation of elasmobranches is complexly organized into many of the same nuclei that are found in frogs, reptiles, birds, and mammals. J. Comp. Neurol. 403:534–560, 1999. © 1999 Wiley-Liss, Inc.     

Brainstem enkephalinergic projections to spinal autonomic nuclei.

The present studies in the rat employed a combined retrograde transport-immunocytochemical technique to determine the origin in the brainstem of enkephalin (Enk) projections to spinal sympathetic nuclei, including the intermediolateralis nucleus, pars principalis (ILp). We found that Enk projections to the ILp nucleus are found in such serotonergic-containing areas as the raphe obscurus; raphe pallidus; gigantocellular reticular nucleus, pars alpha; paragigantocellular lateral nucleus; raphe magnus; and the rostral extension of the raphe magnus nucleus. The adrenergic-containing rostroventrolateral reticular nucleus as well as the noradrenergic-containing areas A5, A7, ventral locus coeruleus, subcoeruleus, and fiber pathway linking the locus coeruleus and A5/A7 send Enk projections to ILp. In the pons, a large contralateral Enk projection to spinal sympathetic nuclei was found medial to the facial nerve and medial to the motor nucleus of the trigeminal nerve. These observations show the existence of a large number of Enk brainstem regions that can influence spinal autonomic centers via descending supraspinal projections.