Peripheral nerve grafts to the frog optic tectum: A morphological study of the axon reaction in trigeminal motor and sensory neurons

The mandibular branch of the trigeminal nerve was severed and the proximal stump was grafted onto the optic tectum in adult Rana pipiens. The resultant changes occurring in the cell bodies of origin in the ipsilateral trigeminal motor and mesencephalic nuclei were studied qualitatively and quantitatively. Nucleolar, nuclear, and somal cross-sectional areas increased in size significantly approximately 3 days after surgery and peaked at 6 weeks postsurgery. This swelling, in which the nucleolus was most severely affected, gradually reversed itself and disappeared by 24 weeks after surgery. Despite the cell enlargement, cytoplasmic basophilia was maintained or even slightly increased. These morphologic changes suggest a strong anabolic reaction. Two differences were found between the motoneurons and the sensory neurons. First, the morphometric cell changes occurred at a faster rate in neurons of the trigeminal motor nucleus than in those of the mesencephalic nucleus. The time course of the motoneuron response correlated well with that of axonal regeneration from the nerve graft. Second, there was a delayed loss of mesencephalic nucleus cells between 12 and 24 weeks after surgery, whereas cells of the trigeminal motor nucleus were maintained at all survival times studied. Taken together with sensory cell loss in the trigeminal ganglion, this suggests a greater viability of regenerating motoneurons.     

Ultrastructure of jaw muscle spindle afferents within the rat trigeminal mesencephalic nucleus

This study examined the ultrastructures of neuronal elements within trigeminal mesencephalic nucleus by labeling masseteric mesencephalic neurons and masseter motoneurons with injection of horseradish peroxidase into masseteric muscle. Of eight horseradish peroxidase-labeled muscle spindle afferents examined, four terminals showed synaptic contact with labeled dendrites of masseteric motoneurons, two with labeled somata, and the remaining two with unlabeled dendrites. A few of the labeled dendrites showed intimate contact with the somata of the trigeminal mesencephalic nucleus neurons. These results provide morphological evidence of synaptic contact of recurring masseteric muscle spindle afferents with the trigeminal mesencephalic nucleus somata and also suggest the presence of electrical synapses between the somata of the trigeminal mesencephalic nucleus neurons and dendrites of jaw-closing motoneurons.     

Developmental regulation of connexins 26, 32, 36, and 43 in trigeminal neurons

The transition from sucking to chewing during postnatal development is accompanied by changes in masticatory muscle activity patterns. We previously demonstrated that changes in numerous parameters of chemical synapses among neurons, and intrinsic membrane properties of neurons, comprising brainstem oral-motor circuits are coincident with changes in masticatory muscle activity patterns. Considering recent findings that implicate a role for gap junctions in early locomotor and respiratory behaviors, our present study focuses on the developmental regulation of connexin proteins in trigeminal neurons as a first step in understanding a role for gap junctions in developing oral-motor circuits used for ingestive behaviors. We conducted immunohistochemistry studies to examine connexin (Cx) 26, 32, 36, and 43 expression in trigeminal motor and mesencephalic trigeminal nuclei during postnatal development at the light and electron microscopic levels. Postnatal days (P) 1, 6, 14, 21, and adult mice were used. Cx32, 36, and 43 expression was developmentally regulated in the trigeminal motor nucleus, while Cx26 expression remained high throughout postnatal development. In the mesencephalic trigeminal nucleus, Cx26, 32, and 43 expression was intense throughout development, with only Cx36 showing a developmental regulation. Ultrastructural examination of neonatal trigeminal motoneurons and mesencephalic trigeminal neurons revealed connexin expression in cell membranes, cytoplasm, and cell nuclei (Cx43, Cx32). Our results show that connexin proteins are differentially regulated between trigeminal motoneurons and mesencephalic trigeminal neurons during development, and suggest a possible role for gap junctions in the development of trigeminal neurons and the function and maturation of oral-motor circuits.     

Afferent projections to the orbicularis oculi motoneuronal cell group. An autoradiographical tracing study in the cat

The motoneurons innervating the orbicularis oculi muscle from a subgroup within the facial nucleus, called the intermediate facial subnucleus. This makes it possible to study afferents to these motoneurons by means of autoradiographical tracing techniques. Many different injections were made in the brainstem and diencephalon and the afferent projections to the intermediate facial subnucleus were studied. The results indicated that these afferents were derived from the following brainstem areas: the dorsal red nucleus and the mesencephalic tegmentum dorsal to it; the olivary pretectal nucleus and/or the nucleus of the optic tract; the dorsolateral pontine tegmentum (parabrachial nuclei and nucleus of K?lliker-Fuse) and principal trigeminal nucleus; the ventrolateral pontine tegmentum at the level of the motor trigeminal nucleus; the caudal medullary medial tegmentum; the lateral tegmentum at the level of the rostral pole of the hypoglossal nucleus and the ventral part of the trigeminal nucleus and the nucleus raphe pallidus and caudal raphe magnus including the adjoining medullary tegmentum. These latter projections probably belong to a general motoneuronal control system. The mesencephalic projections are mainly contralateral, the caudal pontine and upper medullary lateral tegmental projections are mainly ipsilateral and the caudal medullary projections are bilateral. It is suggested that the different afferent pathways subserve different functions of the orbicularis oculi motoneurons. Interneurons in the dorsolateral pontine and lateral medullary tegmentum may serve as relay for cortical and limbic influences on the orbicularis oculi musculature, while interneurons in the ventrolateral pontine and caudal medullary tegmentum may take part in the neuronal organization of the blink reflex.     

An HRP study of the central projections from primary sensory neurons innervating the rat masseter muscle

Retrograde and transganglionic transport of horseradish peroxidase has been used to study the cell bodies of origin and the central projections of neurons innervating the rat masseter muscle. Labeled cell bodies were observed both in the trigeminal ganglion and in the mesencephalic trigeminal nucleus. Major central projections from mesencephalic trigeminal neurons were traced to the supratrigeminal nucleus and to the brainstem reticular formation. Smaller projections from these neurons could be followed to the borders of the solitary tract and hypoglossal nuclei as well as to lamina V of nucleus caudalis and corresponding areas in the dorsal horn at C₁−C₂ spinal cord segments. Labeling from trigeminal ganglion neurons was observed close to the trigeminal tract in all subdivisions of the trigeminal sensory nuclear complex and in the dorsal horn lamina I at C₁ and C₂ levels.     

Mesencephalic innervation of the vibrissal follicle–sinus complex in the mouse embryo

Peripheral projections of neurones whose cell bodies lie in the mesencephalic nucleus of the fifth cranial nerve, situated between the central grey and mesencephalic reticular formation, were studied in mouse embryos aged between day 9 and 15 and in postnatal day 1 mice. Nonspecific neural antibody staining allowed visualisation of the developing cranial nerves, in particular the descending mesencephalic tract. This facilitated successful dissection of the descending mesencephalic tract and trigeminal ganglion in the heads of fresh mouse embryos and postnatal mice. The fluorescent dye, 1,1'-dioctadecyl-3,3,3',3'-tetramethylindocarbocyanine perchlorate (Dil), was injected into the descending mesencephalic tract in mouse embryos aged 12.5, 13.5 and 15 days of gestation and also into postnatal day 1 mice. Following a period of incubation, 100 microm sections were viewed under visible light and episcopic fluorescence. Mesencephalic neurones were observed to pass superiorly over the trigeminal ganglion and enter the maxillary division to innervate vibrissal follicle-sinus complexes, whilst none was observed innervating mandibular and maxillary intraoral structures. There was no fluorescent labelling in non-Dil injected control specimens. Using a highly specific neuronal tracer, this study shows that mesencephalic neurones in the periphery project exclusively to follicle sinus complexes in the developing mouse embryo and remain at least until postnatal day 1. These observations, contrary to those made in other animals, indicate a species specificity of mesencephalic peripheral projections.     

Nitric oxide synthase/nicotinamide adenine dinucleotide phosphate-diaphorase in the brainstem trigeminal nuclei after transection of the masseteric nerve in rats.

In this study, the responses of nicotinamide adenine dinucleotide phosphate-diaphorase (NADPH-d) and neuronal nitric oxide synthase (nNOS) activities were quantitatively analyzed at different times in both ipsilateral and contralateral sides of trigeminal nuclei, after unilateral trigeminal muscle nerve transection, in Sprague Dawley rats. In the control animals, both NADPH-d- and nNOS-positive neurons were constitutively distributed in the rostrolateral solitary tract nucleus, dorsomedial part of trigeminal nucleus oralis (Vo/Sn), and superficial layers (VcI/II) of the trigeminal nucleus caudalis (Vc). NADPH-d-positive neurons appeared in the trigeminal mesencephalic nucleus ipsilaterally at 5 days (mean +/- SEM: 30.5 +/- 5.6) and were maintained until 8 weeks (33 +/- 10.6) after the denervation. In the trigeminal motor nucleus, NADPH-d-positive neurons appeared transiently and bilaterally, peaking at 1 week (663.5 +/- 156.2, ipsilateral side; 687.5 +/- 118.6, contralateral side) after unilateral denervation of the masseteric nerve. In both Vo/Sn and Vc, the number of NADPH-d-positive neurons in the control animals showed a decrease at 3 days but significantly increased from 5 days to 1 week and gradually fell to the control values by 8 weeks after the denervation. There were no significant differences observed between the two sides in either Vo/Sn or Vc. nNOS-positive neurons were similarly distributed and the numbers of labeled neurons were similar to those of NADPH-d-positive neurons after the denervation, although the changes were delayed by approximately 1 week. In conclusion, after unilateral nerve transection, the peak NADPH-d activity occurs 1 week prior to nNOS activity.     

Jaw-muscle spindle afferent pathways to the trigeminal motor nucleus in the rat.

Neural pathways conveying proprioceptive feedback from the jaw muscles were studied in rats by combining retrograde and intracellular neuronal labeling. Initially, horseradish peroxidase was iontophoresed unilaterally into the trigeminal motor nucleus (Vmo). Two days later, 1-5 jaw-muscle spindle afferent axons located in the mesencephalic trigeminal nucleus were physiologically identified and intracellularly stained with biotinamide. Stained mesencephalic trigeminal jaw-muscle spindle afferent axon collaterals and boutons were predominantly distributed in the supratrigeminal region (Vsup), Vmo, dorsomedial trigeminal principal sensory nucleus (Vpdm), parvicellular reticular formation (PCRt), alpha division of the parvicellular reticular formation (PCRtA), and dorsomedial portions of the spinal trigeminal subnuclei oralis (Vodm), and interpolaris (Vidm). Numerous neurons retrogradely labeled with horseradish peroxidase from the trigeminal motor nucleus were found bilaterally in the PCRt, PCRtA, Vodm, and Vidm. Retrogradely labeled neurons were also present contralaterally in the Vsup, Vpdm, Vmo, peritrigeminal zone, and bilaterally in the dorsal medullary reticular field. Putative contacts between intracellularly stained mesencephalic trigeminal jaw-muscle spindle afferent boutons and trigeminal premotor neurons retrogradely labeled with horseradish peroxidase were found in the ipsilateral Vodm, PCRtA, and PCRt, as well as the contralateral Vsup, Vmo, Vodm, PCRt, and PCRtA. Thus, multiple disynaptic jaw-muscle spindle afferent-motoneuron circuits exist. These pathways are likely to convey long-latency jaw-muscle stretch reflexes and may contribute to stiffness regulation of the masticatory muscles.     

A tract-tracing study of the central projections of the mesencephalic nucleus of the trigeminus in the guppy (Lebistes reticulatus, Teleostei), with some observations on the descending trigeminal tract

We studied the central projections of the mesencephalic nucleus of the trigeminal nerve (MesV) in the guppy (Lebistes reticulatus), after application of horseradish peroxidase or fluorescein dextran amine into the eye orbit. A small number (1 to 13) of large mesencephalic trigeminal neurons were solid labelled in the ipsilateral rostral mesencephalon. At the level of the trigeminal nerve entrance, the united process of each mesencephalic trigeminal cell bifurcates, giving rise to a peripheral branch that exits in the trigeminal nerve and a descending branch that runs caudally in a medial bundle separated from the descending trigeminal tract. This bundle passes close to the visceromotor nuclei of the medulla oblongata. Descending processes give rise to short collaterals to the descending nucleus of the trigeminus and the ventrolateral reticular area. Most MesV descending fibres terminate in this ventrolateral field at the transition of the medulla to the spinal cord, but one or two fibres could be followed to the C6 level, where they give rise to collaterals to the dorsal funicular nucleus. No collaterals directed to the trigeminal motor nucleus, the cerebellum, or the mesencephalic tegmentum were observed. These projections were also compared with those of the descending trigeminal tract.     

Enkephalin-, substance P- and serotonin-like immunoreactive axonal varicosities in close apposition to perikarya of mesencephalic trigeminal nucleus neurons in the cat

A light microscopic study in adult cats provided evidence suggesting that neuronal cell bodies of mesencephalic trigeminal nucleus neurons were often in direct contact with axonal varicosities showing enkephalin-, substance P- or serotonin-like immunoreactivity.     

Trigeminal mesencephalic neurons innervating functionally identified muscle spindles and involved in the monosynaptic stretch reflex of the lateral pterygoid muscle of the guinea pig

Location of the neurons in the trigeminal mesencephalic nucleus innervating stretch receptors of the lateral pterygoid muscle and the mode of their synaptic connection on the lateral pterygoid motoneurons of the guinea pig were studied physiologically as well as morphologically, in comparison with the trigeminal mesencephalic neurons innervating muscle spindles in the superficial masseter muscle, with the following results: stimulation of the caudal half of the trigeminal mesencephalic nucleus evoked monosynaptic excitatory postsynaptic potentials in the ipsilateral lateral pterygoid motoneurons. Stimulation of the lateral pterygoid nerve directly evoked spike potentials in the neurons located in the caudal half of the ipsilateral trigeminal mesencephalic nucleus, which responded with increased firing to stretch, and with silent period to twitch, of the ipsilateral lateral pterygoid muscle. Averaging of intracellular potentials of the lateral pterygoid motoneurons with extracellular spike potentials of these trigeminal mesencephalic neurons revealed excitatory postsynaptic potentials after a monosynaptic latency, but no inhibitory postsynaptic potentials. Injection of horseradish peroxidase into the lateral pterygoid muscle labeled 15-20 cells in the caudal half of the ipsilateral trigeminal mesencephalic nucleus, while 174-228 cells retrogradely labeled by horseradish peroxidase were found throughout the whole rostrocaudal extent of the ipsilateral trigeminal mesencephalic nucleus following injection of horseradish peroxidase into the masseter muscle. It was concluded that neurons in the caudal half of the trigeminal mesencephalic nucleus send their peripheral processes to stretch receptors, presumably muscle spindles, in the ipsilateral lateral pterygoid muscle and that their central processes have excitatory synapses on ipsilateral lateral pterygoid motoneurons, thus comprising the afferent limb of a monosynaptic stretch reflex arc of the lateral pterygoid muscle of the guinea pig.     

Cytochemical characteristics of neurons in the trigeminal mesencephalic nucleus of hatchling chicks

The goal of the present study was to identify cytochemical markers characteristic of muscle afferents in hatchling chicks. To this end, we stained neurons in the trigeminal mesencephalic nucleus with a variety of markers that label subsets of neurons in avian dorsal root ganglia. We found that trigeminal mesencephalic neurons are surprisingly heterogeneous in their cytochemical make-up, expressing, to varying degrees, substance P, cholecystokinin, carbonic anhydrase, calbindin D-28k, parvalbumin, and S-100β. Calbindin D28k and S-100β appeared to be expressed equally in medial and lateral divisions of the trigeminal mesencephalic nucleus. In contrast, substance P- and cholecystokinin-immunoreactive neurons were more abundant in the medial division, whereas carbonic anhydrase activity and parvalbumin immunoreactivity were stronger in the lateral division. We were unable to detect met-enkephalin, neuropeptide Y, calcitonin gene-related peptide, vasoactive intestinal peptide, somatostatin, γ-aminobutyric acid, or tyrosine hydroxylase in the trigeminal mesencephalic nucleus. Moreover, these neurons did not appear to bind the lectin Dolichos biflorus agglutinin. The heterogeneity of expression of markers among trigeminal mesencephalic nucleus neurons, especially between neurons in the medial and lateral divisions, suggests that these neurons are functionally diverse.     

Orexin B immunoreactive fibers and terminals innervate the sensory and motor neurons of jaw-elevator muscles in the rat

The relationship between orexinergic terminals and the sensory and motor neurons of jaw-elevator muscles was examined by means of anti-orexin B (OXB) immunohistochemistry combined with horseradish peroxidase (HRP) retrograde tracing in the rat. HRP was initially injected into the jaw-elevator muscles; 48 h later the animals were sacrificed and fixed for HRP reaction and anti-OXB immunohistochemistry. OXB-like terminals were observed to distribute in the trigeminal mesencephalic nucleus (Vme) and the trigeminal motor nucleus (Vmo) where they closely contact the Vme neuronal somata and the Vmo neuronal somata and dendrites retrogradely labeled with HRP. The results of this study provide anatomical evidence of a direct OXB orexinergic innervation of the sensory and motor neurons controlling jaw-elevator muscles involved in mastication. Its functional significance related to the feeding behavior and bruxism is discussed.     

Trigeminal proprioceptive projections to the hypoglossal nucleus and the cervical ventral gray column

Trigeminal proprioceptive projections to the hypoglossal nucleus and the cervical ventral gray column in the cat were investigated by means of neuroanatomical and neurophysiological methods. Degeneration studies (Nauta and Fink-Heimer methods) involved circumscribed electrolytic lesions of the trigeminal mesencephalic nucleus and/or the supratrigeminal nucleus. Degenerated fibers in Probst's tract, which is composed of the central processes of trigeminal mesencephalic neurons, terminated in the ventrolateral portion of the ipsilateral hypoglossal nucleus and in the medial part of the ventral gray column of C₁–C₄. The descending juxtatrigeminal fascicle, a separate bundle of degenerated fibers, originated from the supratrigeminal region, which is known to receive processes from trigeminal mesencephalic neurons. This descending fascicle contributed fibers to the spinal trigeminal nucleus and the juxtatrigeminal reticular formation, from which region interneurons connect to the hypoglossal nucleus. Probst's tract, as well as the descending juxtatrigeminal fascicle, could be considered as parts of two separate polysynaptic pathways from trigeminal proprioceptors to those motoneurons responsible for the innervation of the tongue and infrahyoid musculature. Electrophysiological experiments revealed that proprioceptive muscle afferents from the masseter muscle project directly to the ipsilateral hypoglossal nucleus and to the ipsilateral upper cervical ventral column.     

Profiles of serotonin receptors in the developing human thalamus

The critical importance of the thalamus and its serotonergic innervation with respect to neuropsychiatric syndromes is increasingly recognized. This study investigates the localization of serotonin (5-hydroxytryptamine; 5-HT) receptors by immunohistochemistry in the thalamic nuclei of human fetuses aged 21 to 32 weeks of gestation. Results indicate that, already at 21 weeks of gestation, two 5-HT receptors are present in the dorsomedial nucleus of the developing thalamus: 5-HT2A receptors are localized in neurons and 5-HT2C receptors in fibers. By 31 and 32 weeks of gestation, 5-HT1A and 5-HT4 receptors are also detected in neuronal fibers of the same nucleus. At this later developmental stage, the percentage of 5-HT2A labeled neurons has significantly increased in the dorsomedial nucleus, and 5-HT2C positive neurons are observed in the centromedian and lateroventral thalamic nuclei as well. In contrast, neither neuronal cells nor fibers display any immunoreactivity for 5-HT3 or 5-HT6 receptors at any of the ages examined. Our observation that 5-HT1A, 5-HT2A, 5-HT2C and 5-HT4 receptors are present in the human thalamus prenatally indicates that 5-HT may play a role during fetal development. Disrupted development of the thalamic serotonergic system during this gestational period may contribute to the pathophysiology of neuropsychiatric disorders.     

Developmentally Regulated Expression of mRNA for Neurotrophin High-Affinity (trk) Receptors within Chick Trigeminal Sensory Neurons

To investigate the distribution of neurons within the developing trigeminal sensory system which express mRNA for each of the three known high-affinity neurotrophin receptors (trk, trkB and trkC), we have performed in situ hybridization histochemistry on serial sections through the trigeminal ganglion and trigeminal mesencephalic nucleus at various ages of development using specific antisense oligonucleotide probes. We show that trkC mRNA is first expressed in the chicken embryo at stage 13, in presumptive neurons prior to the formation of the ganglion, that trkB mRNA labelling is initially observed within peripheral neurons slightly later, at stage 19, and that trk mRNA expression is not detectable until around embryonic day 3.5 (stage 21/22). The neurons which exhibit mRNA labelling for each of the high-affinity receptors occupy discrete regions within the ganglion, indicating that the ganglion comprises distinct neuronal subpopulations, each of which has a different capacity to respond to the different neurotrophins. Neurons which express trk mRNA are confined to the proximal region of the ganglion, whereas those which express trkB mRNA and trkC mRNA are located in two distinct regions within the distal aspect and also within the trigeminal mesencephalic nucleus. From the estimation of the number of neurons which exhibit labelling between embryonic days 9 and 18, we determined that the expression of mRNA for the high-affinity receptors changes during embryonic development of the ganglion. This is consistent with the observed differences in the response to neurotrophins in vitro.     

Immunohistochemical localization of calretinin in the rat hindbrain

The localization of calretinin in the rat hindbrain was examined immunohistochemically with antiserum against calretinin purified from the guinea pig brain. Calretinin immunoreactivity was found within neuronal elements. The distribution of calretinin-immunoreactive cell bodies and fibers is presented in schematic drawings and summarized in a table. Major calretinin-immunoreactive neurons were found in the lateral and medial geniculate nuclei, substantia nigra, ventral tegmental area, interpeduncular nucleus, periaqueductal gray, mesencephalic trigeminal nucleus, superior and inferior colliculi, pontine nuclei, parabrachial nucleus, dorsal and laterodorsal tegmental nuclei, cochlear nuclei, vestibular nuclei, medullary reticular nuclei, nucleus of the solitary tract, area postrema, substantia gelatinosa of the spinal trigeminal nucleus, and cerebellum. These results show that distinct calretinin-immunoreactive neurons are widely distributed in the rat hindbrain.     

Appearance of neuronal S-100β during development of the rat brain

In addition to being an astroglial protein, S-100β is localised in distinct populations of neurons in the adult rat hindbrain. We report, here, the expression of S-100β in both neurons and glia of the rat brain during development. Prenatally, S-100β immunoreactivity was confined to glial cells close to the germinal zone. After birth, S-100β positive glial cells were seen mainly in the brainstem and cerebellum, while only a few were detected in cerebral cortex and hippocampus. The number of S-100β containing glial cells increased steadily during the first 2 postnatal weeks after which the adult pattern was attained. No S-100β containing neurons were present prenatally. The first S-100β containing neurons were seen in the mesencephalic trigeminal nucleus at postnatal day 1 (P1), and in the motor trigeminal nucleus at P3. Neuronal S-100β immunoreactivity in other nuclei was mostly attained from the 10th to the 21st postnatal day. The neuronal S-100β immunoreactivity was first detected in the cell nuclei during development, then increased in the cytoplasm with ages. A nuclear staining in many immunoreactive neurons persisted until the adult. It usually took 1 to 2 weeks for neuronal S-100β to attain the adult staining pattern, i.e., heavy staining of the cytoplasm and processes, after its appearance. The forebrain never contained S-100β positive neurons. The S-100β is first expressed in glial cells, suggesting it is primarily of the glial origin. Coupled with neurotrophic effects of the protein, the time course of neuronal S-100β expression during the critical period of neuronal development implies that it may be involved in neuronal differentiation and maturation.     

Distribution of the P2X2 receptor subunit of the ATP-gated ion channels in the rat central nervous system

The distribution of the P2X2 receptor subunit of the adenosine 5'-triphosphate (ATP)-gated ion channels was examined in the adult rat central nervous system (CNS) by using P2X2 receptor-specific antisera and riboprobe-based in situ hybridisation. P2X2 receptor mRNA expression matched the P2X2 receptor protein localisation. An extensive expression pattern was observed, including: olfactory bulb, cerebral cortex, hippocampus, habenula, thalamic and subthalamic nuclei, caudate putamen, posteromedial amygdalo-hippocampal and amygdalo-cortical nuclei, substantia nigra pars compacta, ventromedial and arcuate hypothalamic nuclei, supraoptic nucleus, tuberomammillary nucleus, mesencephalic trigeminal nucleus, dorsal raphe, locus coeruleus, medial parabrachial nucleus, tegmental areas, pontine nuclei, red nucleus, lateral superior olive, cochlear nuclei, spinal trigeminal nuclei, cranial motor nuclei, ventrolateral medulla, area postrema, nucleus of solitary tract, and cerebellar cortex. In the spinal cord, P2X2 receptor expression was highest in the dorsal horn, with significant neuronal labeling in the ventral horn and intermediolateral cell column. The identification of extensive P2X2 receptor immunoreactivity and mRNA distribution within the CNS demonstrated here provides a basis for the P2X receptor antagonist pharmacology reported in electrophysiological studies. These data support the role for extracellular ATP acting as a fast neurotransmitter at pre- and postsynaptic sites in processes such as sensory transmission, sensory-motor integration, motor and autonomic control, and in neuronal phenomena such as long-term potentiation (LTP) and depression (LTD). Additionally, labelling of neuroglia and fibre tracts supports a diverse role for extracellular ATP in CNS homeostasis.     

Morphology of jaw-muscle spindle afferents in the rat

The morphology of jaw-muscle spindle afferents in the rat has been studied by intra-axonal injection of horseradish peroxidase. All stained axons were located in the motor root of the trigeminal nerve and could be traced dorsomedially to the vicinity of the trigeminal motor nucleus, where they divided into an ascending branch in the tract of the mesencephalic nucleus and a descending branch in the tract of Probst. Axon collaterals and swellings on fine collateral branches presumed to be synaptic boutons were located in the following regions: the trigeminal motor nucleus, the region dorsal to the trigeminal motor nucleus including the supratrigeminal nucleus, the parvicellular reticular formation immediately caudal to the trigeminal motor nucleus, the reticular formation at the level of the facial nucleus, and the caudal portion of the mesencephalic nucleus. No evidence of a projection to the cerebellum was observed. Boutons were most numerous in the region surrounding the trigeminal motor nucleus, especially dorsally. Here they were not demonstrated in close proximity to counterstained cells, and therefore it was not possible to determine how many of these contacts are located on cells in this region and how many are on the distal dendrites of trigeminal motorneurons. Boutons located within the trigeminal motor nucleus were always confined to a small portion of the nucleus and were significantly larger than those located dorsally. Some boutons were found in close apposition to trigeminal motorneurons and presumably make somatic contacts. These results suggest that jaw-muscle spindle afferents make somatic and proximal dendritic contacts with only a limited number of trigeminal motorneurons and also project to masticatory interneuronal regions dorsal and caudal to the motor nucleus.