大鼠三叉神经中脑核神经元细胞内HRP标记的研究

本文用电生理方法对大鼠三叉神经中脑核进行定位后将微电极插入该核的神经元,进行细胞内HRP注射。对53个标记神经元的形态、突起的行径、投射范围及其终末分支的特点和终止模式进行了研究。结果表明:(1)三叉神经中脑核神经元有单极、假单极、双极和多极神经元等四种类型,这些神经元有大有小,形状也多样;(2)单极和假单极神经元的突起行程长,投射广泛,侧支极其丰富。它们投射到中间神经元核团(三叉上核、三叉间核,桥延网状结构小细胞网状核等),运动核团(三叉运动核、外展神经副核、面神经副核等)和感觉核团(感觉主核背肉侧部、三叉脊束核吻侧亚核背内侧部等)。此外,还有个别突起投射到动眼神经核、滑车神经核和孤束中。(3)本文首次注意到,这些神经元在性质不同的核群其终末特点和终止模式不同,而在同类核群的各核中其终末特点和终止模式相似。(4)本文还观察到中脑核神经元的周围轴突发出侧支分布到三叉运动核。(5)本文首次提供了三叉神经中脑核神经元向感觉主核背内侧部、外展神经副核和面神经副核投射的资料。此外,本文还就三叉神经本体觉脑内传递通路进行了讨论。     

NEURAL PATHWAYS OF TRIGEMINAL PROPRIOCEPTIVE AFFERENTS COORDINATE ORAL MOTOR BEHAVIORS

Neural pathways and synaptic connections from the trigeminal mesencephalic nucleus (Vme) neurons to the cranial motor nuclei were studied in the rat using double labelling methodologies of intracellular Neurobiotin staining combined with retrograde horseradish peroxidase (HRP) transport, anterograde biotinylated dextran amine (BDA) tracing combined with retrograde HRP transport, and a dual fluorescent labelling of BDA anterograde combined tracing with Cholera Toxin B (CTB) retrograde transport. Direct projections and synapses were demonstrated from Vme neuronal boutons to motoneurons (MNs) of the trigeminal motor nucleus (Vmo), the hypoglossal nucleus (XⅡ) and the ambiguus nucleus (Amb). Indirect projections and pathways from Vme neurons to the cranial motor nuclei including Vmo, XⅡ, the facial nucleus (VⅡ) and the cervical spinal cord (C1～5) were seen to relay on their premotor neurons. The premotor neurons of above cranial motor nuclei were overlapped in bilateral premotor neuronal pool including the parvocellular reticular formation (PCRt) and its alpha division (PCRtA), the dorsomedial part of the spinal trigeminal nucleus oralis (Vodm), and interpolaris (Vidm), the medullary reticular nucleus dorsal division (MdD), the supratrigeminal region (Vsup) and the dorsomedial part of the principal trigeminal sensory nucleus (Vpdm). Synapses between Vme neuronal boutons and Vmo and XⅡ MNs and XII premotor neurons were predominantly asymmetric. There were four types of synaptic organizations, i.e. synaptic convergence; synaptic divergence presynaptic inhibition and afferent feedforward inhibition seen between Vme boutons and Vmo, XⅡ MNs and between Vme boutons and XⅡ premotor neurons. The results of present studies have demonstrated direct pathways from the trigeminal proprioceptive afferents to Vmo, XⅡ and Amb MNs, and indirect pathways from the trigeminal proprioceptive afferents to bilateral Vmo, XⅡ, VⅡ and C1～5 via their premotor neurons. It provides neuroanatomical network to elucidate trigeminal proprioceptive afferents coordinate oral motor behaviors.     

Distribution of nerve growth factor receptor-like immunoreactivity in the adult rat central nervous system. Effect of colchicine and correlation with the cholinergic system--II. Brainstem, cerebellum and spinal cord

Multiple nuclei and fiber tracts in the adult rat brainstem and spinal cord were found to contain nerve growth factor receptor-related protein, as recognized by the monoclonal antibody 192-IgG. Both cholinergic and non-cholinergic sensory and motor regions demonstrated immunoreactive cell bodies and fibers. Nerve growth factor receptor-immunoreactive cells were seen in the mesencephalic nucleus of trigeminal nerve, superior colliculus, parabrachial, prepositus hypoglossal, raphe, dorsal and ventral cochlear, interstitial nucleus of the vestibular nerve, ambiguus and reticular nuclei, cerebellum and ventral spinal cord. Immunoreactive cells resembling neuroglia were distributed subpially along the superior colliculus. Intracerebroventricular injection of colchicine resulted in significantly increased nerve growth factor receptor immunoreactivity in all previously positive neurons and especially in certain neurons of the cochlear and ambiguus nuclei. It also resulted in the visualization of receptor immunoreactivity in certain neurons which were normally non-immunoreactive including cerebellar Purkinje cells, neurons of the central gray, locus coeruleus, facial, dorsal motor vagal and hypoglossal nuclei. In normal animals, nerve growth factor receptor-immunoreactive fibers and varicosities occurred in the trigeminal nerve nuclei, pontine, vestibular, parabrachial, facial, hypoglossal, dorsal motor vagal, solitary, gracile and cuneate nuclei and spinal cord. Although most fiber-like immunoreactive structures were probably axons and nerve terminals, neuroglial or extracellular localizations could not be excluded in some areas. For example, the medial nucleus of the inferior olive and most cerebellar nuclei contained diffuse non-fibrillar receptor immunoreactivity. The presence of nerve growth factor receptor-like immunoreactivity in cell bodies and fibers of several sensory and motor areas of the adult rat brainstem, cerebellum and spinal cord suggests multifocal actions of nerve growth factor or a nerve growth factor-like substance. Although the degree of overlap between nerve growth factor receptor- and choline acetyltransferase-containing regions in the brainstem is not as great as in the forebrain, our findings suggest a potential influence of nerve growth factor or nerve growth factor-like substances on cholinergic systems outside the forebrain. Furthermore, the disparities which occur imply that non-cholinergic nerve growth factor receptor-containing neurons of the brainstem, cerebellum and spinal cord may be affected by such trophic substances.     

大鼠中枢运动核团内P物质受体的定位分布

观察大鼠脑干的躯体运动核,副交感运动核和脊髓前内Ｐ物质受体的定位分布,阐明ＳＰ在运动核团的作用部位。方法免疫组织化学染色技术。结论ＳＰ通过ＳＰ受体的介导在中枢运动核团内对运动神经元发挥调控作用。     

Segregation of lemniscal inputs and motor cortex outputs in cat ventral thalamic nuclei: application of a novel technique

Summary A double labeling method that permits accurate delineation of the terminals of medial lemniscal fibers was used to determine whether thalamic neurons projecting to motor cortex in the cat are in a position to be contacted by such terminals. Thalamic neurons in the VL nucleus were retrogradely labeled by injections of fluorogold placed in the cytoarchitectonically defined area 4, while lemniscal axons and their terminal boutons were anterogradely labeled, in a Golgi-like manner, from injections of Fast Blue placed under physiological control in different parts of the contralateral dorsal column nuclei. In additional experiments, spinothalamic fibers were similarly labeled by injections of Fast Blue in the spinal cord. The results reveal that there is no significant overlap in the distributions of lemniscal terminals and motor cortex-projecting neurons and that no somata or proximal dendrites of motor cortex-projecting neurons are in a position to receive lemniscal terminals. Spinothalamic terminals, on the other hand, end in clusters around motor cortex-projecting neurons in the VL nucleus as well as in other nuclei and are a more likely route for short latency somatosensory inputs to the motor cortex.Abbreviations AD anterodorsal nucleus - AM anteromedial nucleus - AP area postrema - AV anteroventral nucleus - C cuneate nucleus - CeM central medial nucleus - CL central lateral nucleus - CM centre médian nucleus - EC external cuneate nucleus - G gracile nucleus - L limitans nucleus - LD lateral dorsal nucleus - LP lateral posterior nucleus - MGM magnocellular medial geniculate nucleus - MD mediodorsal nucleus - MTT mamillothalamic tract - MV medioventral nucleus - Pc paracentral nucleus - Pf parafascicular nucleus - Po posterior nuclei - R reticular nucleus - RF fasciculus retroflexus - S solitary nucleus - SG suprageniculate nucleus - T spinal trigeminal nucleus - VA ventral anterior nucleus - VIN vestibular nuclei - VL ventral lateral nucleus - VMb basal ventral medial nucleus - VMp principal ventral medial nucleus - VPL ventral posterior lateral nucleus - VPM ventral posterior medial nucleus - ZI zona incerta - 1,2,3a,3b,4 fields of cerebral cortex - C4, C5, C6 spinal cord segments - 5SP,5ST spinal trigeminal nucleus and tract - 10, 12 vagal and hypoglossal nuclei     

Projections from the nucleus of the solitary tract in the rat

The axonal projections of neurons in and near the nucleus of the solitary tract have been visualized using titrated amino acid autoradiography. Axons of neurons of this nucleus ramify extensively within the nucleus itself, but much less so in the nucleus commissuralis. They also enter cranial motor nuclei within the medulla. Axons originating in the anterior part of the nucleus of the solitary tract extend to the hypoglossal, facial and probably trigeminal motor nuclei, but not to the dorsal motor nucleus of the vagus or the nucleus ambiguus. The posterior part of the nucleus of the solitary tract projects to all these motor nuclei. In the spinal cord solitary nucleus axons remain in the medial gray directly caudal to the solitary nucleus itself. The distribution becomes very weak by C₃ after some fibers spread laterally into the caudal trigeminal nucleus. Fibers are labeled in the contralateral ventral columns, but they could not be unequivocably attributed to solitary neurons. Axons ascending from the nucleus of the solitary tract extend no further rostrally than the pons, where they terminate in the caudal end of the parabrachial nuclei.Although often treated as entirely separate systems, the present results indicate that secondary gustatory neurons in the anterior solitary nucleus and secondary visceral afferent neurons in the posterior solitary nucleus have very similar rostral and caudal projections. The pontine parabrachial nuclei, the rostral termination of solitary nucleus neurons, have extensive direct connections to the thalamus, the hypothalamus and the limbic forebrain. Assuming similar connections occur in other mammals, these findings establish the existence of di-synaptic visceral afferent access to the highest autonomic integrative centers in the brain.     

Tachykinin immunoreactivity in terminals of trigeminal afferent fibers in adult and fetal monkey thalamus

Summary Immunocytochemistry of fetal and adult monkey thalamus reveals a dense concentration of tachykinin immunoreactive fibers and terminals in the dorsolateral part of the VPM nucleus in which the contralateral side of the head, face and mouth is represented. The immunoreactive fibers enter the VPM nucleus from the thalamic fasciculus and electron microscopy reveals that they form large terminals resembling those of lemniscal axons and terminating in VPM on dendrites of relay neurons and on presynaptic dendrites of interneurons. Double labeling strategies involving immunostaining for tachykinins after retrograde labeling of brainstem neurons projecting to the VPM failed to reveal the origin of the fibers. The brainstem trigeminal nuclei, however, are regarded as the most likely sources of the VPM-projecting, tachykinin positive fibers.Abbreviations AB ambiguus nucleus - AN abducens nucleus - C cuneate nucleus - CD dorsal cochlear nucleus - CL central lateral nucleus - CM centre médian nucleus - D dendrite - DR dorsal raphe - DV dorsal vagal nucleus - EC external cuneate nucleus - FM medial longitudinal fasciculus - FN facial nucleus - G gracile nucleus - Gc gigantocellular reticular formation - HN hypoglossal nucleus - ICP inferior cerebellar peduncle - IO inferior olivary complex - LC locus coeruleus - LL lateral lemniscus - LM medial lemniscus - M5 motor trigeminal nucleus - NS solitary nucleus - OS superior olivary complex - P dendritic protrusion - Pb parabrachial nucleus - Pc parvocellular reticular formation - PLa anterior pulvinar nucleus - Pp prepositus hypoglossi nucleus - Ps presynaptic region - Py pyramidal tract - P5 principal sensory trigeminal nucleus - R reticular nucleus - RF reticular formation - RL lateral reticular nucleus - S5 spinal trigeminal nucleus - T terminal - T5 spinal trigeminal tract - VL lateral vestibular nucleus - VM medial vestibular nucleus - VMb basal ventral medial nucleus - VPI ventral posterior inferior nucleus - VPL ventral posterior lateral nucleus - VPM ventral posterior medial nucleus - VR ventral raphe - VS superior vestibular nucleus - VSp spinal vestibular nucleus - ZI zona incerta - 5 trigeminal nerve - 6 abducens nerve - 7 facial nerve     

大鼠运动核内5-羟色胺1A、2A、5A受体的定位分布

为了阐明５ 羟色胺在中枢神经系统内与运动神经元结合的精确部位,本研究用免疫细胞化学技术分别观察了大鼠躯体运动核和内脏运动核内５ 羟色胺１Ａ、２Ａ、５Ａ 受体的定位分布。在躯体运动核内观察到：（１）５ 羟色胺１Ａ 受体样阳性神经元和纤维主要分布于动眼神经核、滑车神经核、三叉神经运动核、面神经核、舌下神经核和脊髓前角;（２）５ 羟色胺２Ａ 受体样阳性神经元主要见于动眼神经核、三叉神经运动核、面神经核、舌下神经核和脊髓前角,但阳性纤维和终末却密集地分布于三叉神经运动核、面神经核、舌下神经核和脊髓前角等处,除此之外动眼神经核、滑车神经核、展神经核和疑核内也能见到中等密度的阳性纤维和终末,纤维和终末的分布范围和染色浓度、密度都较神经元为明显;（３）少量淡染的５ 羟色胺５Ａ 受体样阳性神经元和稀疏的阳性纤维及终末主要见于三叉神经运动核、面神经核、舌下神经核和脊髓前角。在内脏运动核内观察的结果是：（１）动眼神经副交感核（Ｅ Ｗ 核）、上涎核、迷走神经背核、骶髓副交感核和胸髓侧角内仅有少量５ 羟色胺１Ａ 受体样阳性神经元、纤维和终末分布;（２）５ 羟色胺２Ａ 受体样阳性神经元和较密集的阳性纤维和终末见于Ｅ Ｗ 核、迷走神经背核、骶?     

Immunohistochemical mapping of neurophysins and calcitonin gene-related peptide in the human brainstem and cervical spinal cord

Our study investigates the distribution of neurophysins (Nph), proteins that are part of the precursors for vasopressin and oxytocin, and calcitonin gene-related peptide (CGRP) in the human brainstem by immunohistochemistry. Both peptides were found in discrete regions of the human hindbrain. Nph could be demonstrated exclusively in fibers and punctate perineural varicosities that were travelling within the mesencephalic central gray, substantia nigra, as well as locus coeruleus, medial longitudinal fascicle, raphe, nucleus of the solitary tract, lateral reticular nucleus and area postrema. A few varicosities were seen in the substantia gelatinosa of the spinal trigeminal tract and its continuation into the dorsal horn of the cervical spinal cord. In contrast to these observations. CGRP-immunoreactive fibers were found to be densest in the spinal tract of the trigeminal nerve and the dorsal horn of the spinal cord. In addition, fibers and varicosities could be demonstrated in numerous distinct brain regions, such as locus coeruleus and subcoeruleus, solitary tract, cuneate nucleus, raphe and periaqueductal gray. CGRP-immunoreactivity was also present in perikarya in the ventral horn of the spinal cord, as well as motor nuclei of cranial nerves, i.e., hypoglossal nucleus, ambiguous nucleus. Our results suggest that Nph-immunoreactivity in the human brainstem may be present predominantly within long fiber projections from hypothalamic neurosecretory nuclei, in analogy to data obtained from rodents, whereas CGRP may play a role in the branchiomotor system as well as in intrinsic or extrinsic projections involved in autonomic regulation and integration of sensory information.     

Projections from the rostral parvocellular reticular formation to pontine and medullary nuclei in the rat: Involvement in autonomic regulation and orofacial motor control

The efferent connections of the rostral parvocellular reticular formation to pontine and medullary nuclei in the rat were studied with anterogradely transported Phaseolus vulgaris leucoagglutinin. Dense innervations from the rostral parvocellular reticular formation were found in the mesencephalic trigeminal nucleus, the supratrigeminal area, the motor trigeminal nucleus, the facial, hypoglossal and parabrachial nuclei and specific parts of the caudal parvocellular reticular formation, including nucleus linearis and the dorsal reticular nucleus of the medulla. The raphe nuclei, nucleus of the solitary tract, inferior olive, dorsal principal sensory, spinal trigeminal nuclei and gigantocellular reticular nucleus and the ventral reticular nucleus of the medulla received moderate projections. In general, the projections from the rostral parvocellular reticular formation were bilateral with an ipsilateral dominance. The dorsal motor vagus and the ambiguus nuclei were not labeled.

It is concluded that the rostral parvocellular reticular formation participates in regulation of orofacial motor control and in neural networks for limbic control of metabolic homeostasis.     

Neuronal expression of Raf protooncogene in the brain stem of adult guinea pig

The Raf protooncogenes encode for cytoplasmic serine/threonine-specific protein kinases which can be activated via growth factor receptors by phosphorylation. Immunohistochemical and Western blotting studies have proven the existence of Raf protein kinases in neurons of the cerebral cortex of rats and guinea pigs. The aim of the present study was to map the immunohistochemical distribution of Raf kinase-like staining in the brain stem of guinea pig. Polyclonal antibodies were used that were raised against a recombinant viral protein in combination with the avidin-biotin-peroxidase system for detection of immunoreactivity. Specificity of the antibodies was tested in Western blotting experiments. Cytoplasmic immunostaining was observed in motor nuclei of hypoglossal, accessory, vagus, facial, trigeminal, abducent, oculomotor and trochlear nerves, and in the nucleus ambiguus, nucleus retroambigualis, lateral vestibular nucleus, mesencephalic nucleus of the trigeminal nerve, the red nucleus, raphe nuclei and reticular formation. Scattered neurons were stained in other sensory nuclei, such as solitary tract nuclei, medial, dorsal and ventral vestibular nuclei and cochlear nuclei. The spinal trigeminal nucleus and the main sensory nucleus of the trigeminal nerve contained few medium-sized immunoreactive cells. In general, staining was mainly somatodendritic; the axonal plexus was not positive. It is concluded, that the widespread neuronal appearance of cytoplasmic Raf kinase suggests an important role in transmission of trophic and growth factor signals in these neurons.     

Co-localization of nitric oxide synthase and choline acetyltransferase in the brain of the goldfish (Carassius auratus)

This work investigates the nitrergic and cholinergic systems in the brain and spinal cord of the goldfish (Carassius auratus). We studied the immunohistochemical localization of antibodies against the neuronal nitric oxide synthase (nNOS) and choline acetyltransferase (ChAT) by bright-field and confocal microscopy. Nitrergic and cholinergic cells were segregated within the telencephalon, in both dorsal and ventral areas, and co-distributed in some nuclei of the diencephalon, mesencephalon, rhombencephalon, and spinal cord. Double-labeling experiments revealed nNOS/ChAT-positive cells in (1) the diencephalon: the preoptic and suprachiasmatic nuclei, the habenula, the dorsal thalamus, and the nucleus of the medial longitudinal fasciculus; (2) the mesencephalon: the optic tectum, the mesencephalic portion of the trigeminal nucleus, the oculomotor and trochlear nuclei, and the Edinger-Westphal nucleus; and (3) the rhombencephalon: the secondary gustatory nucleus, the nucleus isthmi, the lateral lemniscus nucleus, the cerebellum, the reticular formation, different nuclei of the octaval column, the motor zone of the vagal lobe, and the trigeminal, facial, abducens, glosso-pharyngeal, vagal, and hypobranchial motor nuclei. Double-labeled cells were also observed in the spinal motor column. The percentage of double-labeled cells was different in each studied nucleus, indicating a selective distribution pattern. Because double-labeled cells were more abundant in those nuclei involved with sensory and motor physiological processes, we suggest the involvement of both nitric oxide and acetylcholine in these neural functions in fish.     

Proprioceptive afferents survive in the masseter muscle of trkC knockout mice

Peripheral innervation patterns of proprioceptive afferents from dorsal root ganglia and the mesencephalic trigeminal nucleus were assessed in trkC-deficient mice using immunohistochemistry for protein gene product 9.5 and parvalbumin. In trkC knockout mice, spinal proprioceptive afferents were completely absent in the limb skeletal muscles, M. biceps femoris and M. gastrocnemius, as previously reported. In these same animals, however, proprioceptive afferents from mesencephalic trigeminal nucleus innervated masseter muscles and formed primary endings of muscle spindles. Three wild-type mice averaged 35.7 spindle profiles (range: 31–41), six heterozygotes averaged 32.3 spindles (range: 27–41), and four homozygotes averaged 32.8 spindles (range: 26–42). Parvalbumin and Nissl staining of the brain stem showed approximately 50% surviving mesencephalic trigeminal sensory neurons in trkC-deficient mice. TrkC −/− mice (n=5) had 309.4±15.9 mesencephalic trigeminal sensory cells versus 616.5±26.3 the sensory cells in trkC +/+ mice (n=4).These data indicate that while mesencephalic trigeminal sensory neurons are significantly reduced in number by trkC deletion, they are not completely absent. Furthermore, unlike their spinal counterparts, trigeminal proprioceptive afferents survive and give rise to stretch receptor complexes in masseter muscles of trkC knockout mice. This indicates that spinal and mesencephalic trigeminal proprioceptive afferents have different neurotrophin-supporting system during survival and differentiation. It is likely that one or more other neurotrophin receptors expressed in mesencephalic trigeminal proprioceptive neurons of trkC knockout mice compensate for the lack of normal neurotrophin-3 signaling through trkC.     

Proprioceptive afferents survive in the masseter muscle of trkC knockout mice

Peripheral innervation patterns of proprioceptive afferents from dorsal root ganglia and the mesencephalic trigeminal nucleus were assessed in trkC-deficient mice using immunohistochemistry for protein gene product 9.5 and parvalbumin. In trkC knockout mice, spinal proprioceptive afferents were completely absent in the limb skeletal muscles, M. biceps femoris and M. gastrocnemius, as previously reported. In these same animals, however, proprioceptive afferents from mesencephalic trigeminal nucleus innervated masseter muscles and formed primary endings of muscle spindles. Three wild-type mice averaged 35.7 spindle profiles (range: 31-41), six heterozygotes averaged 32.3 spindles (range: 27-41), and four homozygotes averaged 32.8 spindles (range: 26-42). Parvalbumin and Nissl staining of the brain stem showed approximately 50% surviving mesencephalic trigeminal sensory neurons in trkC-deficient mice. TrkC-/- mice (n = 5) had 309.4 +/- 15.9 mesencephalic trigeminal sensory cells versus 616.5 +/- 26.3 the sensory cells in trkC+/+ mice (n = 4). These data indicate that while mesencephalic trigeminal sensory neurons are significantly reduced in number by trkC deletion, they are not completely absent. Furthermore, unlike their spinal counterparts, trigeminal proprioceptive afferents survive and give rise to stretch receptor complexes in masseter muscles of trkC knockout mice. This indicates that spinal and mesencephalic trigeminal proprioceptive afferents have different neurotrophin-supporting system during survival and differentiation. It is likely that one or more other neurotrophin receptors expressed in mesencephalic trigeminal proprioceptive neurons of trkC knockout mice compensate for the lack of normal neurotrophin-3 signaling through trkC.     

The afferent connections of the inferior olivary complex in rats. An anterograde study using autoradiographic and axonal degeneration techniques

Autoradiographic and axonal degeneration techniques were employed to determine the distribution patterns of inferior olivary afferents whose origins were determined using the horseradish peroxidase method.⁷⁰ The Fink-Heimer stain for degenerating axons was used following lesions of the cerebral cortex and spinal cord, while brainstem and cerebellar afferents were mapped by tritiated leucine autoradiography.After unilateral lesions of the mid-thoracic spinal cord, degenerating axons were observed within the subnuclei a and b of the caudolateral medial accessory olive and in the caudolateral dorsal accessory olive. Degeneration after upper cervical cord lesions extended more rostrally and medially within the same olivary subdivisions.Several nuclei within the caudal brainstem projected to the inferior olivary complex. The dorsal column nuclei distributed fibers primarily contralaterally to the lateral part of the dorsal accessory olive and to the caudolateral part of the medial accessory olive; the spinal trigeminal nucleus projected contralaterally to the rostromedial dorsal accessory olive; the medial and inferior vestibular nuclei projected to the ipsilateral subnuclei b, c, and β of the medial accessory olive and to the contralateral dorsomedial cell column; the nucleus prepositus hypoglossi sent fibers to the subnuclei c and β, the dorsal cap and the ventrolateral outgrowth; the lateral reticular nucleus projected to the subnucleus a of the caudolateral medial accessory olive bilaterally; and the reticular formation distributed fibers to the dorsal accessory olive contralaterally and to the β subnucleus ipsilaterally.Study of inferior olivary complex afferents from the deep cerebellar nuclei showed a projection from the fastigial nucleus to the β subnucleus and the ventrolateral outgrowth. The dentate and interpositus nuclei demonstrated topographic connections from these nuclei to the principal olive and accessory olives, respectively. All cerebellar connections were predominantly contralateral.Analysis of mesencephalic and diencephalic areas also demonstrated several inferior olivary complex afferent systems: the caudal pretectum and the superior colliculus projected to the subnucleus c contralaterally and the dorsal lamella of the principal olive ipsilaterally; the nucleus of the optic tract sent fibers to the dorsal cap; the lateral deep mesencephalic nucleus distributed fibers to the ipsilateral dorsal accessory olive and β subnucleus; the medial terminal nucleus of the accessory optic tract projected ipsilaterally to the ventrolateral outgrowth; and several areas including the medial deep mesencephalic nucleus, periaqueductal gray, the nucleus of Darkschewitsch, the subparafascicular nucleus, the rostral red nucleus and the prerubral field all projected ipsilaterally to the principal olive, rostral medial accessory olive, ventrolateral outgrowth and, to a lesser extent, the caudal medial accessory olive, dorsal cap and β subnucleus.Lesions of the frontal cortex produced axonal degeneration primarily ipsilaterally within many olivary subdivisions, especially the medial dorsal accessory olive and the caudomedial medial accessory olive.Although some notable differences in the distribution and laterality of fibers are described, our findings generally corroborate several earlier reports which used different techniques on a variety of species. Inferior olivary afferents from functionally related areas typically demonstrated similar distribution patterns within the subdivisions of the inferior olivary complex. These patterns suggest a functional localization within the inferior olivary complex which may facilitate an understanding of afferents from areas whose functions are not clearly known.     

The afferent connections of the inferior olivary complex in rats: A study using the retrograde transport of horseradish peroxidase

Retrograde transport of horseradish peroxidase (HRP) was used to define the origin of afferents to the inferior olivary complex (IOC) in rats. Using both ventral and dorsal surgical approaches to the brainstem, HRP was injected into the IOC through a micropipette affixed to the tip of a 1-μl Hamilton syringe. After a 2-day postoperative survival, animals were sacrificed by transcardiac perfusion with a 1% paraformaldehyde-1.25% gluteraldehyde solution, and brains were processed according to the DeOlmos protocol (1977), using o-dianisidine as the chromogen. Labeled cells were found at many levels of the nervous system extending from lumbar spinal cord to cerebral cortex. This wide-ranging input from numerous regions clearly underscores the complexity of the IOC and its apparent involvement in several functions. Within the spinal cord, labeled neurons were identified from cervical to lumbar but not at sacral levels. These neurons were found contralaterally in the neck region of the dorsal horn and in the medial portions of the intermediate gray. In the caudal brainstem, reactive cells in the dorsal column nuclei, the spinal trigeminal nucleus, and the subnucleus y of the vestibular complex were observed primarily contralateral to the injection sites. Labeling within the gigantocellular, magnocellular, ventral, and lateral reticular nuclei and the nucleus prepositus hypoglossi was primarily ipsilateral. Reactive neurons in the medial and inferior vestibular nuclei were predominantly ipsilateral or contralateral to HRP injections into the caudal or rostral IOC, respectively. The dentate and interposed nuclei of the cerebellum contained small, lightly labeled neurons primarily contralateral to the injection site, while the fastigial nuclei contained a few relatively large, heavily labeled cells bilateral to caudal olivary injections. Ipsilaterally labeled mesencephalic regions included the periaqueductal gray, interstitial nucleus of Cajal, rostromedial red nucleus, ventral tegmental area, medial terminal nucleus of the accessory optic tract, nucleus of the optic tract, and the lateral deep mesencephalic nucleus. The caudal part of the pretectum and small cells of the stratum profundum of the superior colliculus were labeled predominantly contralateral to the injection. In the caudal diencephalon labeled neurons were most numerous within the nucleus of Darkschewitsch and the subparafascicular nucleus, primarily ipsilateral to olivary injections. Scattered reactive neurons were also found within the ipsilateral zone incerta. With the exception of the zona incerta, all labeled mesencephalic and diencephalic nuclei had some bilateral representation of labeled cells. No labeled neurons were identified within the basal ganglia, while numerous reactive cells were found bilaterally within layer V of the frontal and parietal cerebral cortex.     

大鼠三叉神经中脑核神经元中枢突的行径和分布—HRP跨节追踪研究

将HRP注入大鼠咬肌神经干,在同侧的半月节、咬肌神经核、三叉神经中脑核、三叉上核等处出现标记细胞。大量的跨节标记的中脑核神经元中枢突在同侧Probst束内下降,除一般所记载的Probst束的起始段外,由Ⅶ核水平向尾侧也清楚地看到了此束的范围逐渐缩小且由较疏散的小束组成,位于桥、延网状结构的背外侧部,下可达颈髓上段。此束在走行中,其纤维除终止于咀嚼核、三叉上核外,左面神经核水平有浓密的标记终末止于桥脑网状结构背外侧部的特定部分,形成外、内两簇标记终末群。此区曾有人称为Probst核,但在构筑学上未发现有明确的特征。中脑核神经元中枢突大量终止于此,它有何意义尚有待进一步探索。此外,此束的纤维还分布于孤束核、舌下神经核、颈髓上段后角同Ⅴ-Ⅵ层等处,特别是所有实验例的吻侧疑核处均出现浓密的标记终末。咬肌神经内来自肌梭以外的传入神经元在下颌节的胞体被标记,其中枢突进入脊束背侧部下行,在三叉脊束核尾侧亚核的Ⅰ、Ⅱ重层出现少量标记终末,并有极少量纤维通往Ⅴ、Ⅵ层。     

On the innervation of trigeminal mesencephalic primary afferent neurons by adenosine deaminase-containing projections from the hypothalamus in the rat

The localization and sources of adenosine deaminase-containing structures in the mesencephalic nucleus of the trigeminal nerve of the rat was studied using indirect immunofluorescence or immunoperoxidase immunohistochemical staining techniques for adenosine deaminase in combination with retrograde fluorescent tracing or lesion methods. The majority of large mesencephalic neurons were engulfed by a dense adenosine deaminase-immunoreactive plexus. Immunostaining was often punctate surrounding neuronal profiles or sometimes had the appearance of varicose fibers coursing over the neuronal surface. Occasionally, immunostained axons were found travelling towards and contacting mesencephalic neurons. Mesencephalic neuronal somas surrounded by immunofluorescence staining for adenosine deaminase were simultaneously labelled with fast blue after injections of this dye into the temporalis or masseter muscles of mastication. Injections of fast blue into the mesencephalic nucleus resulted in fast blue labelling of adenosine deaminase-immunoreactive neurons in the tuberal, caudal and postmammillary caudal magnocellular nuclei of the hypothalamus. Ablation of these hypothalamic nuclei caused a near total depletion of adenosine deaminase-immunostained fibers in the mesencephalic nucleus including those associated with mesencephalic neurons. It is concluded that adenosine deaminase-containing neurons in the posterior hypothalamus innervate mesencephalic primary sensory neurons, which are known to convey proprioceptive input to trigeminal motor nuclei controlling jaw muscles. The possibility is considered that the hypothalamus, via a direct action on these sensory neurons, may exert automatic control over jaw movements related to aggressive attack, defensive or feeding behavior. In addition, it appears that mesencephalic neurons may provide an ideal model system for electrophysiological investigations of the neurotransmitter(s) utilized by adenosine deaminase-containing hypothalamic projections.     

Descending brainstem projections of the pedunculopontine tegmental nucleus in the rat

Summary Descending brainstem projections from the pedunculopontine tegmental nucleus (PPN) were studied in the rat by use of the anterograde tracerPhaseolus vulgaris-leucoagglutinin (PHA-L) and the retrograde tracer lectin-conjugated horseradish peroxidase (HRP-WGA). Results of these experiments demonstrated prominent bilateral projections to the pontomedullary reticular nuclei, but direct connections to the motor and sensory nuclei of the cranial nerves could not be ascertained. The PPN fibers terminated mainly in the pontine reticular nuclei oralis and caudalis and in ventromedial portions (pars alpha and pars ventralis) of the gigantocellular reticular nucleus. A smaller number of labeled fibers distributed to more dorsal regions of the gigantocellular nucleus, lateral paragigantocellular, ventral reticular nucleus of the medulla and lateral reticular nucleus. Although a significant number of PHA-L labeled fibers was seen in two cases in the contralateral medial portion of the facial nucleus, and all cases exhibited a sparse predominantly ipsilateral projection to the lateral facial motor neurons, the retrograde tracing experiments have revealed that these facial afferents originated in the nuclei surrounding the PPN. The results are discussed in the context of PPN involvement in motor functions. It is suggested that the PPN may participate in a complex network involved in the orienting reflex.Abbreviations Am ambiguus nucleus - AP area postrema - Ac 7 accessory facial nucleus - asc 7 ascending fibers, facial nerve - CG central gray - Cnf cuneiform nucleus - Cu cuneate nucleus - cp cerebral peduncle - g7 germ facial nerve - Gi gigantocellular reticular nucleus - GiA gigantocellular reticular nucleus, pars alpha - GiV gigantocellular reticular nucleus, pars ventralis - Gr gracile nucleus - IC inferior colliculus - icp inferior cerebellar peduncle - IO inferior olive - IRt intermediate reticular nucleus - KF Kölliker-Fuse nucleus - LC locus coeruleus - ll lateral lemniscus - vsc ventral spinocerebellar tract - xscp decussation of superior cerebellar peduncle - 3 oculomotor nucleus - 4 trochlear nucleus - 6 abducens nucleus - 5n trigeminal nerve - 7 facial nucleus - 7n facial nerve - 10 dorsal motor nucleus of vagus - 12 hypoglossal nucleus - MPB medial parabrachial nucleus - MVe medial vestibular nucleus - PCRt parvicellular reticular nucleus - PN pontine nucleus - PPNe pedunculopontine tegmental nucleus, pars compacta - PPNd pedunculopontine tegmental nucleus, pars dissipata - Pr5 principal sensory trigeminal nucleus - py pyramidal tract - pyx pyramidal decussation - Rmes mesencephalic reticular nucleus - RN red nucleus - RPc reticularis pontis caudalis nucleus - Rpo reticularis pontis oralis nucleus - RR retrorubral nucleus - RRF retrorubral field rs rubrospinal tract - SC superior colliculus - scp superior cerebellar peduncle - LPB lateral parabrachial nucleus - LPGi lateral paragigantocellular reticular nucleus - LRt lateral reticular nucleus - LSO lateral superior olive - LVe lateral vestibular nucleus - MdD medullary reticular nucleus, dorsal - MdV medullary reticular nucleus, ventral - Me5 mesencephalic trigeminal nucleus - me5 mesencephalic trigeminal tract - ml medial lemniscus - mlf medial longitudinal fasciculus - Mo5 motor trigeminal nucleus - SNc substantia nigra, pars compacta - SNr substantia nigra, pars reticulata - SO superior olive - Sol nucleus of the solitary tract - sol solitary tract - sp5 spinal trigeminal tract - Sp5o spinal trigeminal nucleus, pars oralis - SPTg subpeduncular tegmental nucleus - SpVe spinal vestibular nucleus - Tz nucleus of the trapezoid body - tz trapezoid body - VLL ventral nucleus of the lateral lemniscus This paper is dedicated to Professor Fred Walberg on the occasion of his 70th birthday.     

Jaw-muscle spindle afferent feedback to the cervical spinal cord in the rat

Putative synaptic contacts between masticatory-muscle spindle afferents and brainstem neurons which project to the cervical spinal cord were studied in rats by combining retrograde and intracellular neuronal labeling. Spinal cord projecting neurons were retrogradely labeled via injection of horseradish peroxidase unilaterally or bilaterally into cervical spinal cord segments C2 through C5. Twenty-four hours after the injection of horseradish peroxidase, one to five jaw-muscle spindle afferent axons were physiologically identified and intracellularly stained with biotinamide on each side of the brainstem. Horseradish-peroxidase-labeled neurons were found bilaterally in the supratrigeminal region, trigeminal principal sensory nucleus, parvicellular reticular nucleus including its alpha division, spinal trigeminal subnuclei oralis and interpolaris and the medullary reticular formation. Retrogradely labeled neurons were most numerous in the spinal trigeminal subnucleus oralis, parvicellular reticular formation and the ventral part of the spinal trigeminal subnucleus interpolaris. A small number of horseradish-peroxidase-labeled neurons were also present in the trigeminal mesencephalic nucleus and spinal trigeminal subnucleus caudalis. Appositions between jaw-muscle spindle afferent boutons and spinal projecting neurons were found in the supratrigeminal region, dorsomedial portions of the trigeminal principal sensory nucleus and spinal trigeminal subnuclei oralis and interpolaris, and the parvicellular reticular formation including its alpha division. Putative synaptic contacts were most frequent in the parvicellular reticular formation and the dorsomedial portion of the trigeminal subnucleus oralis. These results indicate that some orofacial proprioceptive feedback transmitted via the mesencephalic trigeminal nucleus reaches the cervical spinal cord directly and suggests that jaw-muscle spindle afferent feedback reaches the cervical spinal cord predominately via relays in the dorsomedial part of the spinal trigeminal subnucleus oralis and the parvicellular reticular formation. It is hypothesized that these pathways are primarily involved in the coordination of jaw and neck movement during mastication and biting. Materials and methods 27 January 1999 / Accepted: 9 May 1999