Whisker-related circuitry in the trigeminal nucleus principalis: ultrastructure.

Trigeminal (V) nucleus principalis (PrV) is the requisite brainstem nucleus in the whisker-to-barrel cortex model system that is widely used to reveal mechanisms of map formation and information processing. Yet, little is known of the actual PrV circuitry. In the ventral "barrelette" portion of the adult mouse PrV, relationships between V primary afferent terminals, thalamic-projecting PrV neurons, and gamma-aminobutyric acid (GABA)-ergic terminals were analyzed in the electron microscope. Primary afferents, thalamic-projecting cells, and GABAergic terminals were labeled, respectively, by Neurobiotin injections in the V ganglion, horseradish peroxidase injections in the thalamus, and postembedding immunogold histochemistry. Primary afferent terminals (Neurobiotin- and glutamate-immunoreactive) display asymmetric and multiple synapses predominantly upon the distal dendrites and spines of PrV cells that project to the thalamus. Primary afferents also synapse upon GABAergic terminals. GABAergic terminals display symmetric synapses onto primary afferent terminals, the somata and dendrites (distal, mostly) of thalamic-projecting neurons, and GABAergic dendrites. Thus, primary afferent inputs through the PrV are subject to pre- and postsynaptic GABAergic influences. As such, circuitry exists in PrV "barrelettes" for primary afferents to directly activate thalamic-projecting and inhibitory local circuit cells. The latter are synaptically associated with themselves, the primary afferents, and with the thalamic-projecting neurons. Thus, whisker-related primary afferent inputs through PrV projection neurons are pre- and postsynaptically modulated by local circuits.

A novel neurological phenotype in mice lacking mitochondrial manganese superoxide dismutase.

Reactive oxygen species (ROS) have been implicated in a wide range of degenerative processes including amyotrophic lateral sclerosis, ischemic heart disease, Alzheimer disease, Parkinson disease and aging. ROS are generated by mitochondria as the toxic by-products of oxidative phosphorylation, their energy generating pathway. Genetic inactivation of the mitochondrial form of superoxide dismutase in mice results in dilated cardiomyopathy, hepatic lipid accumulation and early neonatal death. We report that treatment with the superoxide dismutase (SOD) mimetic Manganese 5, 10, 15, 20-tetrakis (4-benzoic acid) porphyrin (MnTBAP) rescues these Sod2tm1Cje(-/-) mutant mice from this systemic pathology and dramatically prolongs their survival. The animals instead develop a pronounced movement disorder progressing to total debilitation by three weeks of age. Neuropathologic evaluation reveals a striking spongiform degeneration of the cortex and specific brain stem nuclei associated with gliosis and intramyelinic vacuolization similar to that observed in cytotoxic edema and disorders associated with mitochondrial abnormalities such as Leighs disease and Canavans disease. We believe that due to the failure of MnTBAP to cross the blood brain barrier progressive neuropathology is caused by excessive mitochondrial production of ROS. Consequently, MnTBAP-treated Sod2tm1Cje(-/-) mice may provide an excellent model for examining the relationship between free radicals and neurodegenerative diseases and for screening new drugs to treat these disorders.

Ultrastructural analysis of glutamate-immunopositive synapses onto the rat jaw-closing motoneurons during postnatal development.

The excitatory synapses on the jaw-closing (JC) motoneurons mediate the neuronal input that ensures smooth and rhythmic movements of the jaw. Recently, we have shown that the neurotransmitter phenotype of the inhibitory boutons onto JC motoneurons shifts from GABA to glycine, and new inhibitory synapses onto JC motoneurons are continuously formed during postnatal development (Paik et al. [2007] J. Comp. Neurol. 503:779­789). To test whether the developmental pattern of the excitatory synapses onto JC motoneurons differs from that of the inhibitory synapses, we studied the distribution of glutamate-immunopositive boutons onto the rat JC motoneurons during postnatal development by using a combination of retrograde labeling with horseradish peroxidase (HRP), postembedding immunogold staining, and quantitative ultrastructural analysis. The analysis of 175, 281, and 465 boutons contacting somata of JC motoneurons at postnatal days P2, P11, and P31, respectively, revealed that the number of glutamate-immunopositive (Glut(+)) boutons increased by 2.6 times from P2 to P11 and showed no significant change after that, whereas the length of apposition of these boutons increased continuously from P2 to P31, suggesting that the time course for the development of Glut(+) boutons differed from that for Glut(-) boutons, most of which were immunopositive for GABA and/or glycine. Our findings indicate that excitatory and inhibitory synapses onto JC motoneurons exhibit distinctly different developmental patterns that may be closely related to the maturation of the masticatory system.

Development of γ-aminobutyric acid-, glycine-, and glutamate-immunopositive boutons on the rat genioglossal motoneurons.

Detailed information about the development of excitatory and inhibitory synapses on the genioglossal (GG) motoneuron may help to understand the mechanism of fine control of GG motoneuron firing and the coordinated tongue movement during postnatal development. For this, we investigated the development of γ-aminobutyric acid (GABA)-immunopositive (GABA +), glycine + (Gly +), and glutamate + (Glut +) axon terminals (boutons) on the somata of rat GG motoneurons at a postnatal day 2 (P2), P6 and P18 by retrograde labeling of GG motoneurons with horseradish peroxidase, electron microscopic postembedding immunogold staining with GABA, Gly, and Glut antisera, and quantitative analysis. The number of boutons per GG motoneuron somata and the mean length of bouton apposition, measures of bouton size and synaptic covering percentage, were significantly increased from P2/P6 to P18. The number and fraction of GABA + only boutons of all boutons decreased significantly, whereas those of Gly + only boutons increased significantly from P2/P6 to P18, suggesting developmental switch from GABAergic to glycinergic synaptic transmission. The fraction of mixed GABA +/Gly + boutons of all boutons was the highest among inhibitory bouton types throughout the postnatal development. The fractions of excitatory and inhibitory boutons of all boutons remained unchanged during postnatal development. These findings reveal a distinct developmental pattern of inhibitory synapses on the GG motoneurons different from that on spinal or trigeminal motoneurons, which may have an important role in the regulation of the precise and coordinated movements of the tongue during the maturation of the oral motor system.

Upregulation of silent information regulator 1 alleviates mitochondrial dysfunction in the trigeminal nucleus caudalis in a rat model of chronic migraine.

Although the mechanism of chronic migraine is still unclear, more and more studies have shown that mitochondrial dysfunction plays a possible role in migraine pathophysiology. Silent information regulator 1 (SIRT1) plays a vital role in mitochondrial dysfunction in many diseases. However, there is no research on the role of SIRT1 in mitochondrial dysfunction of chronic migraine. The aim of this study was to explore the role of SIRT1 in mitochondrial dysfunction in chronic migraine. A rat model was established through repeated dural infusions of inflammatory soup for 7 days to simulate chronic migraine attacks. Cutaneous hyperalgesia caused by the repeated infusions of inflammatory soup was detected using the von Frey test. Then, we detected SIRT1 expression in the trigeminal nucleus caudalis. To explore the effect of SIRT1 on mitochondrial dysfunction in chronic migraine rats, we examined whether SRT1720, an activator of SIRT1, altered mitochondrial dysfunction in chronic migraine rats. Repeated infusions of inflammatory soup resulted in cutaneous hyperalgesia accompanied by downregulation of SIRT1. SRT1720 significantly alleviated the cutaneous hyperalgesia induced by repeated infusions of inflammatory soup. Furthermore, activation of SIRT1 markedly increased the expression of peroxisome proliferator-activated receptor gamma-coactivator 1-alpha, transcription factor A, nuclear respiratory factor 1 and nuclear respiratory factor 2 mitochondrial DNA and increased the ATP content and mitochondrial membrane potential. Our results indicate that SIRT1 may have an effect on mitochondrial dysfunction in chronic migraine rats. Activation of SIRT1 has a protective effect on mitochondrial function in chronic migraine rats.

Ultrastructural analysis of glutamate-, GABA-, and glycine-immunopositive boutons from supratrigeminal premotoneurons in the rat trigeminal motor nucleus.

The supratrigeminal region (Vsup) is important for coordination of smooth jaw movement. However, little is known about the synaptic connections of the Vsup premotoneurons with the trigeminal motor neurons. In the present study, we examined axon terminals of Vsup premotoneurons in the contralateral trigeminal motor nucleus (Vmo) by a combination of anterograde tracing with cholera toxin B-horseradish peroxidase (CTB-HRP), postembedding immunohistochemistry for the amino acid transmitters glutamate, GABA, and glycine, and electron microscopy. Tracer injections resulted in anterograde labeling of axon terminals of the Vsup premotoneurons in the motor trigeminal nucleus (Vmo). The labeled boutons in Vmo exhibited immunoreactivity for glutamate, GABA, or glycine: glutamate-immunopositive boutons (69%) were more frequently observed than GABA- or glycine-immunopositive boutons (19% and 12%, respectively). Although most labeled boutons (97%) made synaptic contacts with a single postsynaptic dendrite, a few glutamate-immunopositive boutons (3%) showed synaptic contact with two dendrites. No labeled boutons participated in axoaxonic synaptic contacts. Most labeled boutons (78%) were presynaptic to dendritic shafts, and the remaining 22% were presynaptic to somata or primary dendrites. A large proportion of GABA- or glycine-immunopositive boutons (40%) were presynaptic to somata or primary dendrites, whereas most glutamate-immunopositive boutons (86%) were presynaptic to dendritic shafts. These results indicate that axon terminals of Vsup premotoneurons show simple synaptic connection with Vmo neurons. This may provide the anatomical basis for the neural information processing responsible for jaw movement control.

Distribution of excitatory and inhibitory axon terminals on the rat hypoglossal motoneurons.

Detailed information about the excitatory and inhibitory synapses on the hypoglossal motoneurons may help understand the neural mechanism for control of the hypoglossal motoneuron excitability and hence the precise and coordinated movements of the tongue during chewing, swallowing and licking. For this, we investigated the distribution of GABA-, glycine (Gly)- and glutamate (Glut)-immunopositive (+) axon terminals on the genioglossal (GG) motoneurons by retrograde tracing, electron microscopic immunohistochemistry, and quantitative analysis. Small GG motoneurons (< 400 µm2 in cross-sectional area) had fewer primary dendrites, significantly higher nuclear/cytoplasmic ratio, and smaller membrane area covered by synaptic boutons than large GG motoneurons (> 400 µm2). The fraction of inhibitory boutons (GABA + only, Gly + only, and mixed GABA +/Gly + boutons) of all boutons was significantly higher for small GG motoneurons than for large ones, whereas the fraction of Glut + boutons was significantly higher for large GG motoneurons than for small ones. Almost all boutons (> 95%) on both small and large GG motoneurons were GABA + , Gly + or Glut + . The frequency of mixed GABA +/Gly + boutons was the highest among inhibitory boutons types for both small and large GG motoneurons. These findings may elucidate the anatomical substrate for precise regulation of the motoneuron firing required for the fine movements of the tongue, and also suggest that the excitability of small and large GG motoneurons may be regulated differently.

Expression of P2X3 receptor in the trigeminal sensory nuclei of the rat.

Trigeminal primary afferents expressing P2X(3) receptor are involved in the transmission of orofacial nociceptive information. However, little is known about their central projection pattern and ultrastructural features within the trigeminal brainstem sensory nuclei (TBSN). Here we use multiple immunofluorescence and electron microscopy to characterize the P2X(3)-immunopositive (+) neurons in the trigeminal ganglion and describe the distribution and synaptic organization of their central terminals within the rat TBSN, including nuclei principalis (Vp), oralis (Vo), interpolaris (Vi), and caudalis (Vc). In the trigeminal ganglion, P2X(3) immunoreactivity was mainly in small and medium-sized somata, but also frequently in large somata. Although most P2X(3) (+) somata costained for the nonpeptidergic marker IB4, few costained for the peptidergic marker substance P. Most P2X(3) (+) fibers in the sensory root of trigeminal ganglion (92.9%) were unmyelinated, whereas the rest were small myelinated. In the TBSN, P2X(3) immunoreactivity was dispersed in the rostral TBSN but was dense in the superficial laminae of Vc, especially in the inner lamina II. The P2X(3) (+) terminals contained numerous clear, round vesicles and sparse large, dense-core vesicles. Typically, they were presynaptic to one or two dendritic shafts and also frequently postsynaptic to axonal endings, containing pleomorphic vesicles. Such P2X(3) (+) terminals, showing glomerular shape and complex synaptic relationships, and those exhibiting axoaxonic contacts, were more frequently seen in Vp than in any other TBSN. These results suggest that orofacial nociceptive information may be transmitted via P2X(3) (+) afferents to all TBSN and that it may be processed differently in different TBSN.

GABA- and glycine-like immunoreactivity in axonal endings presynaptic to the vibrissa afferents in the cat trigeminal interpolar nucleus.

The goal of this study was to analyze the synaptic interaction of primary afferents with GABA- and/or glycine-immunopositive presynaptic endings in the cat trigeminal interpolar nucleus (Vi). Fast adapting vibrissa afferents were labeled by intra-axonal injections of horseradish peroxidase. Postembedding immunogold labeling on serially cut ultrathin sections and quantitative ultrastructural analysis of the labeled boutons and their presynaptic endings (p-endings) in the Vi were performed. The majority of p-endings presynaptic to labeled boutons (83%) were immunopositive for both GABA and glycine and 8% were immunopositive for glycine alone. A small fraction of p-endings were immunopositive for GABA alone (4%) or immunonegative for both GABA and glycine (4%). Ultrastructural parameters related to synaptic release, i.e. bouton volume, mitochondrial volume, and active zone area, were significantly larger in the labeled boutons of primary afferents than in the p-endings. The volume of labeled boutons was positively correlated with the number of the postsynaptic dendrites and p-endings. In addition, fairly large-sized labeled boutons and p-endings were frequently observed in the Vi. These results reveal that large majority of vibrissa afferents in the Vi are presynaptically modulated by interneurons immunopositive for both GABA and glycine, and suggest that the Vi plays a distinct role in the processing of orofacial sensory information, different from that of other trigeminal sensory nuclei.

Dynorphin-A immunoreactive terminals on the neuronal somata of rat mesencephalic trigeminal nucleus.

Dynorphin-A-like immunoreactivity was investigated in the rat mesencephalic trigeminal nucleus (Mes 5) at the light and electron microscopic levels. Dynorphin-A immunoreactive fibers and puncta, likely representing nerve terminals, were observed throughout rostrocaudal extension of the Mes 5 at the light microscopic level. Within the rostrocaudal extension, more abundant fibers and puncta were localized in the midbrain-pontine junction and pontine areas than in the midbrain area. At the electron microscopic level, dynorphin-A immunoreactive synapses were observed on the somata of Mes 5. Dynorphin-A-like immunoreactivity tended to be restricted to dense-cored vesicles in the synapses. These results suggest that dynorphin-A-containing fiber systems affect mastication through the Mes 5.

Changes in the axon terminals of primary afferents from a single vibrissa in the rat trigeminal nuclei after active touch deprivation or exposure to an enriched environment.

Lasting modifications of sensory input induce structural and functional changes in the brain, but the involvement of primary sensory neurons in this plasticity has been practically ignored. Here, we examine qualitatively and quantitatively the central axonal terminations of a population of trigeminal ganglion neurons, whose peripheral axons innervate a single mystacial vibrissa. Vibrissa follicles are heavily innervated by myelinated and unmyelinated fibers that exit the follicle mainly through a single deep vibrissal nerve. We made intraneural injections of a mixture of cholera-toxin B (CTB) and isolectin B4, tracers for myelinated and unmyelinated fibers, respectively, in three groups of young adult rats: controls, animals subjected to chronic haptic touch deprivation by unilateral whisker trimming, and rats exposed for 2 months to environmental enrichment. The regional and laminar pattern of terminal arborizations in the trigeminal nuclei of the brain stem did not show gross changes after sensory input modification. However, there were significant and widespread increases in the number and size of CTB-labeled varicosities in the enriched condition, and a prominent expansion in both parameters in laminae III-IV of the caudal division of the spinal nucleus in the whisker trimming condition. No obvious changes were detected in IB4-labeled terminals in laminae I-II. These results show that a prolonged exposure to changes in sensory input without any neural damage is capable of inducing structural changes in terminals of primary afferents in mature animals, and highlight the importance of peripheral structures as the presumed earliest players in sensory experience-dependent plasticity.

Expression of vesicular glutamate transporter 1 immunoreactivity in peripheral and central endings of trigeminal mesencephalic nucleus neurons in the rat.

The major neuronal components of the trigeminal mesencephalic nucleus (Vmes) are primary afferent neurons that convey proprioceptive information from the cranioorofacial regions. In the present study, we examined expression of vesicular glutamate transporters (VGLUTs), VGLUT1 and VGLUT2, in the primary afferent neurons of the Vmes (Vmes neurons) in neonatal and adult rats. VGLUT1 immunoreactivity was detected in the cell bodies of Vmes neurons in neonatal rats younger than 11 days old, but not in older rats. However, in situ hybridization signals for VGLUT1 mRNA were detected in both neonatal and adult rats. No VGLUT2 immunoreactivity was detected in Vmes neurons of neonatal or adult rats. VGLUT1 immunoreactivity was also seen in the peripheral sensory endings on the equatorial regions of intrafusal fibers of muscle spindles in the masseter muscles in both neonatal and adult rats. In adult rats injected with cholera toxin B subunit (CTb) into the masseter nerve, central axon terminals of Vmes neurons were identified on masseter motoneurons within the trigeminal motor nucleus (Vm) by transganglionically and retrogradely transported CTb. VGLUT1-immunopositive axon terminals in close apposition to CTb-labeled Vm motoneurons were also detected by dual-immunofluorescence histochemistry for VGLUT1/CTb. Electron microscopy after dual immunolabeling for VGLUT1/CTb by the VGLUT1/immunoperoxidase and CTb/immunogold-silver methods further revealed synaptic contact of VGLUT1- and CTb-immunopositive axon terminals upon CTb-labeled neuronal profiles within the Vm. These data indicate that VGLUT1 is expressed in both the central axon terminals and the peripheral sensory endings of Vmes neurons, although no VGLUT1 immunoreactivity was detectable in the cell bodies of Vmes neurons in adult rats.

Central distribution of nociceptive intradental afferent nerve fibers in the rat.

The central distribution of intradental afferent nerve fibers was investigated by combining electron microscopic observations with a selective method for inducing degeneration of the A delta- and C-type afferent fibers. Degenerating terminals were found on the proprioceptive mesencephalic trigeminal neurons and on dendrites in the neuropil of the trigeminal motor nucleus after application of capsaicin to the rat's lower incisor tooth pulp. The results give anatomical evidence of new sites of central projection of intradental A delta- and C-type fibers whereby the nociceptive information from the tooth pulp can affect jaw muscle activity.

The synaptic microcircuitry associated with primary afferent terminals in the interpolaris and caudalis of trigeminal sensory nuclear complex.

Previous ultrastructural studies indicating a higher number of axoaxonic contacts on individual low-threshold mechanoreceptive afferents in the principalis (Vp) than in the oralis (Vo) of cat trigeminal sensory nuclear complex (TSNC) suggest that the synaptic microcircuitry associated with primary afferents manifests unique differences across the sensory nuclei of TSNC. To address this issue, we analyzed synaptic microcircuits associated with fast adapting vibrissa afferent terminals in the interpolaris (Vi) and caudalis (Vc, laminae III/IV) by using intraaxonal injections of horseradish peroxidase (HRP) in cats. Forty-two and 65 HRP-labeled boutons were analyzed in the Vi and Vc, respectively. The labeled boutons contained clear, spherical vesicles. They most frequently formed asymmetric axodendritic synapses and were commonly postsynaptic to unlabeled axon terminals containing pleomorphic vesicles (p-endings) with symmetric junctions. The examination of synaptic contacts over the entire surface of individual boutons indicated that the afferent boutons made contacts with an average of two postsynaptic targets in the Vi and Vc. In contrast, axoaxonic contacts, and labeled boutons participating in synaptic triads, where p-endings contacted both the boutons and their postsynaptic targets, were, on average, higher in the Vi than in the Vc. These results suggest that the output of sensory information conveyed through low-threshold mechanoreceptive afferents is more strongly controlled at the level of the first synapse by presynaptic and postsynaptic mechanisms in the Vi responsible for sensory discriminative functions than in the Vc for sensorimotor reflexive functions.

Morphology and synaptic connections of myelinated primary axons in the ventrolateral region of rat trigeminal nucleus oralis.

Neurons in the ventrolateral (VL) subdivision of rat trigeminal nucleus oralis (Vo) have most of their dendritic arbors confined within this region. This study examines the morphology and synaptic connections of a population of myelinated primary trigeminal axons that arborize within VL and are in a position to provide input directly to VL neurons. Primary axons were visualized for light and electron microscopic analysis by injecting 30% horseradish peroxidase (HRP) in 2% dimethylsulfoxide (DMSO) into the sensory root of the trigeminal nerve and allowing 24-36 hours for the anterograde transport of HRP into the terminal axonal arbors. This population is characterized by its cone-shaped terminal arbors, which generate many axonal endings (2-8 micron in diameter) along unmyelinated terminal strands. These arbors arise from collaterals emanating from thinly myelinated (2-5 micron in diameter) parent branches descending in the spinal V tract, which, on the basis of their size, are considered to be small myelinated (A sigma) primary trigeminal axons. HRP-labeled P endings belonging to this population of primary axons are scalloped, filled with spherical to ovoid (40-70 nm in diameter) synaptic vesicles, and lie centrally in glomeruli where they make asymmetrical axodendritic synapses on dendritic shafts and spine heads. It is at these synapses that this population of primary trigeminal axons is probably transferring its input directly to the dendritic arbors of VL neurons. The dendritic shafts and spine heads also receive symmetrical to intermediate axodendritic synapses from endings containing flattened (70 X 29 nm) synaptic vesicles. These terminals also establish axo-axonic synapses on the P ending. Other synaptic components found less often in the glomeruli include small terminals containing oval (14-23 nm) synaptic vesicles that establish symmetrical to intermediate synapses on the P ending, boutons containing pleomorphic (35-80 nm) synaptic vesicles that form symmetrical to intermediate synapses on the P ending as well as on dendritic shafts, and small peripheral endings containing round (20-40 nm) synaptic vesicles that establish asymmetrical synapses on dendritic shafts.

Subnuclear organization of the ophidian trigeminal motor nucleus. II. Ultrastructural measurements on motoneurons innervating antagonistic muscles.

Horseradish peroxidase (HRP), injected intramuscularly, specifically labeled motoneurons innervating antagonistic jaw muscles in the cottonmouth mocassin, Agkistrodon piscivorus piscivorus. Adductor mandibulae profundus, part 3a, motoneurons were localized in the lateral regions of the ventral and intermediate subnuclei of the trigeminal (V) motor nucleus. These were large cells containing fine, granular reaction product characteristic of alpha-motoneurons. Small cells, which contained large coarse reaction granules characteristic of gamma-motoneurons were localized in a separate cluster in the lateral regions of the dorsal subnucleus of the V motor nucleus. Depressor mandibulae motoneurons were localized in the ventromedial regions of the facial (VII) motor nucleus, primarily in the caudal half. Cell sizes ranged from 30--50 micrometers in diameter and HRP staining characteristics were variable, indicating a mixed population of motoneuron functional types without the segregation noted in the V motor nucleus. Boutons which made synaptic contact with labeled somata or processes were classified according to morphological type and their frequency of appearance. Boutons containing spherical vesicles (S-, C-, T-) were distributed similarly on motoneurons of both muscles, but more F-boutons, those with flattened vesicles, synapsed on the adductor motoneurons. Comparison of snake bouton distribution with that in mammalian spinal cord indicates that synaptology on all these motoneurons are remarkably similar. The more frequent occurrence of C-boutons, those with a subsynaptic cistern, on reptilian motoneurons may indicate a stronger intrasegmental input, as determined from mammalian degeneration studies.

A light and electron microscopic study of premotor neurons for the trigeminal motor nucleus.

Premotor neurons sending their axons to the trigeminal motor nucleus were observed in the cat by light and electron microscopy after labeling the neurons retrogradely or anterogradely with horseradish peroxidase (HRP). After HRP injection into the trigeminal motor nucleus, retrogradely labeled neurons were seen most frequently in the parvocellular reticular formation bilaterally. Many labeled neurons were also seen contralaterally in the intermediate zone at the rostralmost levels of the cervical cord and its rostral extension into the caudalmost levels of the medulla oblongata. Additionally, some neurons were labeled ipsilaterally in the mesencephalic trigeminal nucleus, contralaterally in the main sensory trigeminal nucleus and the trigeminal motor nucleus, and bilaterally in the oral and interpolar subnuclei of the spinal trigeminal nucleus. Only a few labeled neurons were seen in the confines of the gigantocellular reticular formation. All labeled neurons were small or of medium size; no large neurons were labeled. After HRP injection into the regions around the trigeminal motor nucleus or the parvocellular reticular formation, axodendritic terminals containing HRP granules were found contralaterally within the trigeminal motor nucleus. Some of these labeled terminals were filled with round synaptic vesicles and others contained pleomorphic synaptic vesicles. The varied morphology of labeled axon terminals was considered to reflect the functional heterogeneity of the premotor neurons for the trigeminal motor nucleus.

Axon morphology of mesencephalic trigeminal neurons in a snake, Thamnophis sirtalis.

The morphology of single axons of mesencephalic trigeminal neurons (Mes V) was studied in the eastern garter snake (Thamnophis sirtalis) by solid filling them with an extracellular horseradish peroxidase technique. Each Mes V axon can be divided into central, peripheral, and descending branches. The central branch descends from its soma of origin in the mid-brain to the dorsal aspect of the motor nucleus of the trigeminal (Motor V) and the motor root, where it splits into peripheral and descending branches. The descending branch travels caudally from Motor V to the brainstem-spinal cord junction. The peripheral branch passes dorsal to motor V and joins the motor root of V to exit the brainstem. All three branches issue a massive collateral system that distributes terminal swelling within the nuclear boundaries of Motor V. Single Mes V axons diverge to sparsely contact a large number of motoneurons throughout the nucleus, suggesting that single motoneurons receive a convergent input from many Mes V neurons. Since Motor V contains multiple, highly overlapping motor pools, single afferents are positioned to contact different motor pools. The descending branch is situated medial and adjacent to the spinal sensory nucleus of the trigeminal (Sensory V). It issues a collateral field to the entire length of Sensory V. The terminal swellings of these collaterals form rostrocaudally aligned sheets, flattened in the horizontal plane. Single terminal sheets have a divergent projection to a large field of sensory cells and single, fusiform sensory cells are positioned to receive a convergent projection from many terminal sheets. The results provide the first detailed description of Mes V axon morphology. The overall pattern of these axons closely resembles that recently described for spinal Ia afferent fibers in cat. There is evidence in both cases for divergence of single afferent terminal fields to set of spatially overlapping motor pools and a convergence of input to single motoneurons from a large population of afferents. This anatomical pattern is consistent with the recently proposed role of sensory feedback in the activity of single motoneurons.

Fine structural survey of the rat's brainstem sensory trigeminal complex.

The fine structural organization of the principal sensory trigeminal nucleus was compared with that of the spinal trigeminal nucleus (subnuclei oralis, interpolaris, and the deep layers of caudalis) in adult albino rats. Direct comparisons indicate similarities between all of the subdivisions of the brainstem trigeminal complex both in the major morphological classes of neurons present and in basic patterns of synaptic connections. Major differences between the several subdivisions occur in the relative numbers and distribution of the different cell types. The spinal trigeminal nucleus is distinguished by more numerous large (22-40 micron) polygonal neurons which give rise to long straight primary dendrites. Both the perikaryal surface and the thick primary dendrites of many of these cells are densely innervated by synaptic terminals. Especially large cells of this type are a prominent feature of subnucleus oralis. By contrast, the principal sensory nucleus is distinguished by its high density of small to medium-sized (8-20 micron) round or ovoid neurons. These smaller neurons tend to receive a sparse axosomatic innervation. In addition to these differences the spinal trigeminal neuropil is distinguished by the striking manner in which it is broken up by large rostrocaudally oriented bundles of myelinated axons. Proximal dendrites of polygonal and fusiform neurons often wrap around these large axon bundles. Morphologically heterogeneous populations of synaptic terminals with round vesicles (R terminals) and terminals with predominantly flattened vesicles (F terminals) occur in all of the subdivisions of the trigeminal complex. Both types of terminal make primarily axodendritic synapses, but both also make axosomatic synapses, and axospinous synapses with somatic as well as dendritic spines. In addition, axoaxonic synaptic contacts from F terminals onto large R terminals are seen in all subdivisions. Convincing examples of presynaptic dendrites were not observed in any of the brainstem subdivisions. Synaptic glomeruli, characteristic groupings of dendrites and synaptic terminals, are found throughout the brainstem trigeminal complex. The dendritic elements in these glomeruli tend to be small-diameter dendrites, spines, and large, spinelike appendages. Within the glomerulus these elements are postsynaptic to a single large R terminal and may also be postsynaptic to smaller F terminals. In addition, axoaxonic synaptic contacts from the F terminals onto the R terminal are a consistent feature of trigeminal synaptic glomeruli.(ABSTRACT TRUNCATED AT 400 WORDS)

Morphology and synaptic connections of slowly adapting periodontal afferent terminals in the trigeminal subnuclei principalis and oralis of the cat.

Previous studies suggest that sensory information from primary afferent fibers is processed in a distinct manner in the individual subnuclei of trigeminal sensory nuclear complex. The present study has addressed this issue by using intra-axonal labeling with horseradish peroxidase to examine the ultrastructure and synaptic organization of axon terminals from slowly adapting (SA) periodontal afferents in the ventral subdivision (Vpv) of principalis and the rostro-dorsomedial part (Vo.r) of oralis. Our observations are based on complete or near-complete reconstructions of 139 synaptic boutons in Vpv and 105 in Vo.r. All the labeled boutons contained clear, spherical, synaptic vesicles and were presynaptic to unlabeled dendrites, and they were frequently postsynaptic to unlabeled axon terminals containing pleomorphic synaptic vesicles (P-endings). The P-endings frequently formed axodendritic synapses on dendrites which received axodendritic synapses from labeled boutons (synaptic triads). On the basis of the number of contacts, synaptic arrangements associated with the labeled boutons could be subgrouped into simple (one or two contacts), intermediate (three or four contacts), and complex (five or more contacts) types. The labeled boutons varied from round to elongated forms with smooth to more irregular or scalloped contours. The boutons with scalloped contour were much more frequent in the complex type. The boutons of the intermediate type were significantly smaller than the complex type and larger than the simple type. The SA periodontal afferent terminals participated in each type of synaptic arrangements in Vpv, but were mostly of the simple type in Vo.r. The size of labeled boutons was significantly larger in Vpv than in Vo.r. The total number of axodendritic and axoaxonic contacts per labeled bouton was significantly higher in Vpv than in Vo.r. Another difference was the more frequent occurrence of synaptic triads in Vpv than in Vo.r. These observations provide evidence that sensory information from primary afferent fibers is processed in a different manner in the two subnuclei.