鸣禽中脑听觉核团边缘区神经投射的研究

对鸣禽白腰文鸟（Ｌｏｎｃｈｕｒａ ｓｔｒｉａｔａ）中脑听觉核团－背外侧核（ｎｕｃｌｅｕｓ ｍｅｓｅｎｃｅｐｈａｌｉｃｕｓ ｌａｔｅｒａｌｉｓ,ｐａｒｓ ｄｏｒｓａｌｉｓ,ＭＬｄ）和丘间核（ｎｕｃｌｅｕｓ ｉｎｔｅｒｃｏｌｌｉｃｕｌａｒｉｓ,ＩＣｏ）的脑啡肽免疫化学特异和神经联系进行了研究,结果发现：脑啡肽能标记纤维或细胞主要分布于中脑听觉核团的痛外侧核边缘区、丘间核,而背外侧核中央几乎无分布。双向     

鸣禽中脑听觉核团边缘区神经投射的研究

对鸣禽白腰文鸟(Lonchura striata)中脑听觉核团--背外侧核(nucleus mesencephalicus lateralis, pars dorsalis, MLd) 和丘间核(nucleus intercollicularis, ICo)的脑啡肽免疫化学特性和神经联系进行了研究.结果发现:脑啡肽能标记纤维或细胞主要分布于中脑听觉核团的背外侧核边缘区、丘间核,而背外侧核中央区几乎无分布.双向神经示踪剂生物素结合的葡聚糖胺注射入MLd边缘区后,从端脑到延髓的许多核团或脑区均获得了传出传入神经标记.MLd边缘区接受5个核团或脑区的投射,即:端脑古纹状体粗核(robust archistriatum, RA)壳区、下丘脑视前核(nucleus p reopticus anterior, POA)、脑桥臂旁核(nucleus parabrachialis ventrolateralis, PBv l)、延髓下橄榄上核(nucleus infraolivaris superior,IOS)和延髓喙腹外侧核(nucleus r ostral ventrolateral medulla, RVL); 而MLd边缘区发出纤维,支配间脑卵圆核(nucleus ovoidalis, Ov)壳区和外侧下丘脑(nucleus lateralis hypothalami,LHy).结果提示鸣禽中脑背外侧核边缘区在听觉发声、内分泌活动及其他生理活动的联系中可能具有重要作用.     

Afferent pathways to the region of the vestibular nuclei that participates in cardiovascular and respiratory control

Prior experiments have shown that a region of the medial and inferior vestibular nuclei contributes to cardiovascular and respiratory regulation. In addition to labyrinthine inputs, the majority of neurons in this region of the vestibular nuclei receive signals from the skin, muscle, and viscera, although the pathways conveying these nonlabyrinthine inputs to the vestibular nucleus neurons are unknown. To gain further insight into the afferent pathways to this functionally distinct subdivision of the vestibular complex, we combined monosynaptic mapping with viral transneuronal tracing in the ferret. First order afferent projections were defined by retrograde transport of the beta-subunit of cholera toxin (CTbeta), and the extended polysynaptic circuitry was defined in the same animals by injection of a recombinant of pseudorabies virus Bartha (PRV) into the contralateral vestibular nuclei. Neurons containing CTbeta or infected by retrograde transneuronal transport and replication of PRV were distributed throughout the spinal cord, but were 10 times more prevalent in the cervical cord than the lumbar cord. The labeled spinal neurons were most commonly observed in Rexed's laminae IV-VI and the dorsal portions of laminae VII-VIII. Both the CTbeta and PRV injections also resulted in labeling of neurons in all four vestibular nuclei, the prepositus hypoglossi, the reticular formation, the inferior olivary nucleus, the medullary raphe nuclei, the spinal and principal trigeminal nuclei, the facial nucleus, and the lateral reticular nucleus. Following survival times >/=3 days, PRV-infected neurons were additionally present in nucleus solitarius and the gracile and cuneate nuclei. These data show that an anatomical substrate is present for somatosensory and visceral inputs to influence the activity of cells in the autonomic region of the vestibular nuclei and suggest that these signals are primarily transmitted through brainstem relay neurons.     

Descending pathways from hypothalamus to dorsal motor vagus and ambiguus nuclei in the rat

The anatomical pathways between the hypothalamus and cell groups of the lower medulla that are involved in the neural control of endocrine pancreas activity were investigated. As part of this control system the descending pathways originating from lateral, dorsomedial and ventromedial hypothalamic nuclei towards the dorsal motor vagus and ambiguus nuclei, were studied by retrograde transport of horseradish peroxidase. Very small injections of the tracer, by means of the iontophoretic delivery method, were placed in the dorsal motor vagus, ambiguus and solitary tract nucleus as well as in the various nuclei of the medullary reticular formation. Subsequent retrograde labeling was studied in the hypothalamus and the brainstem. The appearance of considerable retrograde labeling in mesencephalic periventricular grey and rostral mesencephalic reticular formation indicated a possible role for these structures as intermediates in an indirect hypothalamo-medullary control circuitry. This led us to extend the peroxidase injections to these mesencephalic areas after which the hypothalamus was investigated for retrograde labeling. All data combined indicated the existence of three descending pathways, direct and indirect, between hypothalamus and the parasympathetic motor nuclei of the lower medulla.     

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

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

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

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

Optogenetic approaches to characterize the long-range synaptic pathways from the hypothalamus to brain stem autonomic nuclei

Recent advances in optogenetic methods demonstrate the feasibility of selective photoactivation at the soma of neurons that express channelrhodopsin-2 (ChR2), but a comprehensive evaluation of different methods to selectively evoke transmitter release from distant synapses using optogenetic approaches is needed. Here we compared different lentiviral vectors, with sub-population-specific and strong promoters, and transgenic methods to express and photostimulate ChR2 in the long-range projections of paraventricular nucleus of the hypothalamus (PVN) neurons to brain stem cardiac vagal neurons (CVNs). Using PVN subpopulation-specific promoters for vasopressin and oxytocin, we were able to depolarize the soma of these neurons upon photostimulation, but these promoters were not strong enough to drive sufficient expression for optogenetic stimulation and synaptic release from the distal axons. However, utilizing the synapsin promoter photostimulation of distal PVN axons successfully evoked glutamatergic excitatory post-synaptic currents in CVNs. Employing the Cre/loxP system, using the Sim-1 Cre-driver mouse line, we found that the Rosa-CAG-LSL-ChR2-EYFP Cre-responder mice expressed higher levels of ChR2 than the Rosa-CAG-LSL-ChR2-tdTomato line in the PVN, judged by photo-evoked currents at the soma. However, neither was able to drive sufficient expression to observe and photostimulate the long-range projections to brainstem autonomic regions. We conclude that a viral vector approach with a strong promoter is required for successful optogenetic stimulation of distal axons to evoke transmitter release in pre-autonomic PVN neurons. This approach can be very useful to study important hypothalamus-brainstem connections, and can be easily modified to selectively activate other long-range projections within the brain.     

Parallel auditory pathways: projection patterns of the different neuronal populations in the dorsal and ventral cochlear nuclei

The cochlear nuclear complex gives rise to widespread projections to nuclei throughout the brainstem. The projections arise from separate, well-defined populations of cells. None of the cell populations in the cochlear nucleus projects to all brainstem targets, and none of the targets receives inputs from all cell types. The projections of nine distinguishable cell types in the cochlear nucleus—seven in the ventral cochlear nucleus and two in the dorsal cochlear nucleus—are described in this review. Globular bushy cells and two types of spherical bushy cells project to nuclei in the superior olivary complex that play roles in sound localization based on binaural cues. Octopus cells convey precisely timed information to nuclei in the superior olivary complex and lateral lemniscus that, in turn, send inhibitory input to the inferior colliculus. Cochlear root neurons send widespread projections to areas of the reticular formation involved in startle reflexes and autonomic functions. Type I multipolar cells may encode complex features of natural stimuli and send excitatory projections directly to the inferior colliculus. Type II multipolar cells send inhibitory projections to the contralateral cochlear nuclei. Fusiform cells in the dorsal cochlear nucleus appear to be important for the localization of sounds based on spectral cues and send direct excitatory projections to the inferior colliculus. Giant cells in the dorsal cochlear nucleus also project directly to the inferior colliculus; some of them may convey inhibitory inputs to the contralateral cochlear nucleus as well.     

Descending serotonergic, peptidergic and cholinergic pathways from the raphe nuclei: a multiple transmitter complex

The localization of serotonergic, various peptidergic and possibly cholinergic neurons in the medullary raphe nuclei that project to the lumbosacral spinal cord have been studied using a retrograde transport method combined with immunocytochemical and histochemical techniques. Spinally projecting neurons stained for serotonin-like, substance P-like, enkephalin-like and thyrotropin-releasing hormone-like immunoreactivity and for the histochemical marker acetylcholinesterase were all observed in each of the raphe nuclei of the medulla, as well as in the adjacent ventrolateral reticular formation. The similar distributions of the descending serotonergic and peptidergic neurons in the raphe nuclei as well as quantitative data on their relative numbers suggest that a large fraction of raphe-spinal neurons contain serotonin co-existing with one or more peptides in the same cell.     

GABA in the intralaminar thalamic nuclei modulates dopamine release from the two dopaminergic nigro-striatal pathways in the Cat

Halothane-anaesthetized cats implanted with several push-pull cannulae were used to study the effects of the unilateral application of GABA (10(-5) M, 30 min) into thalamic intralaminar nuclei on the in vivo release of (3H)-dopamine [3H)-DA) newly synthesized from (3H)-tyrosine in both caudate nuclei (CN) and substantiae nigrae (SN). GABA applied into the left centralis lateralis nucleus (CL) elicited symmetric changes in the two CN, (3H)-DA release being initially reduced and thereafter markedly increased. On the contrary, an asymmetric pattern of responses was observed in the two CN during and after GABA application into the left centrum medianum parafascicular complex (CM/PF) since (3H)-DA release was decreased ipsilaterally and enhanced contralaterally. Changes in (3H)-DA release intervening in the two SN appeared to be triggered by processes independant from those operating in the CN. Hence, an immediate increase in (3H)-DA release with a time course distinct from that observed in other structures occurred in the contralateral SN after the application of GABA either into the left CL or the left CM/Pf. Furthermore in both cases, a biphasic response (decrease followed by an increase) similar to that intervening in the CN following GABA application into the CL was seen in the ipsilateral SN. Autoradiographic studies performed with (14C)-GABA (10(-5) M) revealed that the amino-acid did not spread out from the site of application. During and after GABA application (10(-5) M, 30 min) into the CL or the CM/Pf neuronal firing was markedly enhanced not only locally but also into respective contralateral homologous structures. These results further confirm the important and specific roles of distinct thalamic nuclei in the bilateral regulation of DA release from dendrites and nerve terminals of the nigro-striatal dopaminergic neurons.     

Topographical organization of pathways from somatosensory cortex through the pontine nuclei to tactile regions of the rat cerebellar hemispheres

The granule cell layer of the cerebellar hemispheres contains a patchy and noncontinuous map of the body surface, consisting of a complex mosaic of multiple perioral tactile representations. Previous physiological studies have shown that cerebrocerebellar mossy fibre projections, conveyed through the pontine nuclei, are mapped in registration with peripheral tactile projections to the cerebellum. In contrast to the fractured cerebellar map, the primary somatosensory cortex (SI) is somatotopically organized. To understand better the map transformation occurring in cerebrocerebellar pathways, we injected axonal tracers in electrophysiologically defined locations in Sprague-Dawley rat folium crus IIa, and mapped the distribution of retrogradely labelled neurons within the pontine nuclei using three-dimensional (3-D) reconstructions. Tracer injections within the large central upper lip patch in crus IIa-labelled neurons located centrally in the pontine nuclei, primarily contralateral to the injected side. Larger injections (covering multiple crus IIa perioral representations) resulted in labelling extending only slightly beyond this region, with a higher density and more ipsilaterally labelled neurons. Combined axonal tracer injections in upper lip representations in SI and crus IIa, revealed a close spatial correspondence between the cerebropontine terminal fields and the crus IIa projecting neurons. Finally, comparisons with previously published three-dimensional distributions of pontine neurons labelled following tracer injections in face receiving regions in the paramedian lobule (downloaded from http://www.rbwb.org) revealed similar correspondence. The present data support the coherent topographical organization of cerebro-ponto-cerebellar networks previously suggested from physiological studies. We discuss the present findings in the context of transformations from cerebral somatotopic to cerebellar fractured tactile representations.     

Identification of the midbrain locomotor nuclei and their descending pathways in the teleost carp, Cyprinus carpio

In order to identify the mesencephalic spinal pathways for initiation of swimming in the carp, we employed electrical and chemical microstimulation of the mesencephalic tegmentum. Electrical stimulation of the midbrain in decerebrate carp produced bilateral or unilateral rhythmic movements of the tail. Bilateral alternating movements were induced by stimulation with the lowest threshold currents to the brain region just beneath the third ventricle at the level of the mid mesencephalon. The region included the nucleus of medial longitudinal fasciculus (Nflm), the medial longitudinal fasciculus (flm), the red nucleus (Nrb). To specify the nuclei of the origin of the descending pathway, we microinjected 0.1 M

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-glutamic acid to the region. Both bilateral and unilateral tail movements were induced, the majority being the latter. The unilateral movements were accompanied with tail flips toward the ipsilateral side of stimulation sites. The smallest injection volume required for initiation of the movement was recorded at the Nflm. Bilateral tail movements were produced only by injections into the medial region between the nucleus of the both sides. The present results imply a crucial role of Nflm neurons in the initiation of swimming Nflm neurons on one side project through flm to the ipsilateral spinal cord along its entire length and regulate activities of the individual central pattern generators.     

Complementary subicular pathways to the anterior thalamic nuclei and mammillary bodies in the rat and macaque monkey brain

The origins of the hippocampal (subicular) projections to the anterior thalamic nuclei and mammillary bodies were compared in rats and macaque monkeys using retrograde tracers. These projections form core components of the Papez circuit, which is vital for normal memory. The study revealed a complex pattern of subicular efferents, consistent with the presence of different, parallel information streams, whose segregation appears more marked in the rat brain. In both species, the cells projecting to the mammillary bodies and anterior thalamic nuclei showed laminar separation but also differed along other hippocampal axes. In the rat, these diencephalic inputs showed complementary topographies in the proximal–distal (columnar) plane, consistent with differential involvement in object‐based (proximal subiculum) and context‐based (distal subiculum) information. The medial mammillary inputs, which arose along the anterior–posterior extent of the rat subiculum, favoured the central subiculum (septal hippocampus) and the more proximal subiculum (temporal hippocampus). In contrast, anterior thalamic inputs were largely confined to the dorsal (i.e. septal and intermediate) subiculum, where projections to the anteromedial nucleus favoured the proximal subiculum while those to the anteroventral nucleus predominantly arose in the distal subiculum. In the macaque, the corresponding diencephalic inputs were again distinguished by anterior–posterior topographies, as subicular inputs to the medial mammillary bodies predominantly arose from the posterior hippocampus while subicular inputs to the anteromedial thalamic nucleus predominantly arose from the anterior hippocampus. Unlike the rat, there was no clear evidence of proximal–distal separation as all of these medial diencephalic projections preferentially arose from the more distal subiculum.     

Proprioceptive pathways to posterior parietal areas MIP and LIPv from the dorsal column nuclei and the postcentral somatosensory cortex

The posterior parietal cortex (PPC) serves as an interface between sensory and motor cortices by integrating multisensory signals with motor-related information. Sensorimotor transformation of somatosensory signals is crucial for the generation and updating of body representations and movement plans. Using retrograde transneuronal transfer of rabies virus in combination with a conventional tracer, we identified direct and polysynaptic somatosensory pathways to two posterior parietal areas, the ventral lateral intraparietal area (LIPv) and the rostral part of the medial intraparietal area (MIP) in macaque monkeys. In addition to direct projections from somatosensory areas 2v and 3a, respectively, we found that LIPv and MIP receive disynaptic inputs from the dorsal column nuclei as directly as these somatosensory areas, via a parallel channel. LIPv is the target of minor neck muscle-related projections from the cuneate (Cu) and the external cuneate nuclei (ECu), and direct projections from area 2v, that likely carry kinesthetic/vestibular/optokinetic-related signals. In contrast, MIP receives major arm and shoulder proprioceptive inputs disynaptically from the rostral Cu and ECu, and trisynaptically (via area 3a) from caudal portions of these nuclei. These findings have important implications for the understanding of the influence of proprioceptive information on movement control operations of the PPC and the formation of body representations. They also contribute to explain the specific deficits of proprioceptive guidance of movement associated to optic ataxia.