Inputs from identified jaw-muscle spindle afferents to trigeminothalamic neurons in the rat: A double-labeling study using retrograde HRP and intracellular biotinamide

Projections from physiologically identified jaw-muscle spindle afferents onto trigeminothalamic neurons were studied in the rat. Trigeminothalamic neurons were identified by means of retrograde transport of horseradish peroxidase from the ventroposteromedial nucleus of the thalamus. Labeled neurons were found contralaterally in the supratrigeminal region (Vsup), the trigeminal principal sensory nucleus, the ventrolateral part of the trigeminal subnucleus oralis, the spinal trigeminal subnuclei interpolaris and caudalis, the reticular formation, and an area ventral to the trigeminal motor nucleus (Vmo) and medial to the trigeminal principal sensory nucleus (AVM). Jaw-muscle spindle afferents were physiologically identified by their increased firing during stretehing of the jaw muscles and intracellularly injected with biotinamide. Axon collaterals and boutons from jaw-muscle spindle afferents were found in Vmo; Vsup; the dorsomedial part of the trigeminal principal sensory nucleus (Vpdm); the dorsomedial part of the spinal trigeminal subnuclei oralis, interpolaris (Vidm) and caudalis; the parvicellular reticular formation (PCRt); and the mesencephalic trigeminal nucleus. Trigeminothalamic neurons in Vsup, Vpdm, Vidm, PCRt, and AVM were associated with axon collaterals and boutons from intracellularly stained jaw-muscle spindle afferents. Trigeminothalamic neurons in Vsup, Vpdm, Vidm, and PCRt were closely apposed by one to 14 intracellularly labeled boutons from jaw-muscle spindle afferents, suggesting a powerful input to some trigeminothalamic neurons. These data demonstrate that muscle length and velocity feedback from jaw-muscle spindle afferents is projected to the contralateral thalamus via multiple regions of the trigeminal system and implicates these pathways in the projection of trigeminal proprioceptive information to the cerebral cortex. © 1995 Wiley-Liss, Inc.     

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.     

The anatomical evidence of recurrent axonal collaterals of the thalamus projecting neurons of the rostral pole of the trigeminal sensory nuclear complex in the rat.

Thalamus projecting neurons and their recurrent axonal collaterals were observed in the dorsomedial part of the trigeminal principal sensory nucleus (Vpdm) and the caudolateral part of supratrigeminal nucleus (Vsup CL) after injection of horseradish peroxidase (HRP) into the contralateral ventrobasal complex of the thalamus (VBm) by using the HRP retrogradely tracing-Golgi-like staining method. About 7% (8/120) parent axons of the labeled cells gave rise to recurrent axon collaterals. However, no retrogradely labeled cells were observed in the VBm after injection of HRP into the Vpdm and Vsup CL. In an electron microscopic study, the terminals of recurrent axon collaterals made synapses with the dendrites of the thalamus projecting neurons or non-labeled neurons in the neuropil of the Vpdm and Vsup CL. It is suggested that the recurrent axon collaterals might play a role of negative feedback in transmission of the proprioceptive message from the jaw-closing muscle spindles to the thalamus.     

Projection of jaw-muscle spindle afferents to the caudal brainstem in rats demonstrated using intracellular biotinamide

Intracellular staining with biotinamide was used to study the axonal projection and synaptic morphology of rat jaw-muscle spindle afferents. Intracellular recordings in the mesencephalic trigeminal nucleus (Vme) were identified as spindle afferent responses by their increased firing during stretching of the jaw-elevator muscles. Biotinamide-stained axon collaterals with boutons were found in the trigeminal motor nucleus (Vmo), Vme, the region dorsal to Vmo including the supratrigeminal region, the dorsomedial portion of the trigeminal principal sensory nucleus, and the dorsomedial part of the rostral spinal trigeminal subnucleus oralis. Additional, previously undescribed projections of jaw-muscle spindle afferents were found to the dorsomedial portiori of the caudal spinal trigeminal subnucleus oralis (Vodm), the dorsomedial part of the spinal trigeminal subnucleus interpolaris (Vidm), the caudal parvicellular reticular formation, laminae IV and V of the spinal trigeminal subnucleus caudalis (Vc), and the dorsal division of the medullary reticular field. Labeled spindle boutons in Vodm formed predominately axodendritic synapses. Some of these boutons received presynaptic inputs from unlabeled P-type boutons containing clear, spherical, or flattened vesicles. In Vidm, labeled collaterals and boutons were densely clustered into glomerular-like structures. Labeled boutons in Vidm made axodendritic, axosomatic, and axoaxonic synapses and received synaptic contacts from unlabeled boutons containng clear, spherical, or flat and pleomorphic vesicles. Unlabeled presynaptic boutons in Vidrn occasionally contained dense core vesicles. Labeled boutons in Vc mainly formed synaptic contacts with large diameter dendrites. This projection of jaw-muscle spindle afferents to caudal brainstem regions may play a significant role in masticatory-muscle stretch reflexes and in the integration of trigeminal proprioceptive information and its transmission to higher centers. © 1995 Wiley-Liss, Inc.     

Projections from subnucleus oralis of the spinal trigeminal nucleus to contralateral thalamus via the relay of juxtatrigeminal nucleus and dorsomedial part of the principal sensory trigeminal nucleus in the rat.

Following injection of HRP into contralateral thalamus, retrogradely labeled cells were observed in principal sensory trigeminal nucleus (Vp) and an area of juxtatrigeminal nucleus (JX) formerly described by John and Tracey (1987). When PHA-L was delivered to dorsomedial part of the subnucleus oralis (Vodm), PHA-L labeled terminals were seen in dorsomedial part of the Vp (Vpdm) and in the JX region. Comparing the distribution of PHA-L labeled terminal field with that of HRP labeled JX neurons showed that the labeled terminals and neurons were overlapped closely in the JX. The distribution patterns of the labeled terminals and JX neurons were also the same: viewed on the coronal planes caudal-rostrally, both of the labelings began to appear at the levels where the facial nerve root was just broken. Rostrally, at middle levels of the motor trigeminal nucleus (Vmo), the labelings showed their typical view covering dorsal and ventral JX (dJX, vJX). The labelings disappeared at rostral poles of the Vmo and Vp. When injections of PHA-L into the Vodm and HRP into the contralateral thalamus was made in one rat, the contacts between Vodm projecting terminals labeled with PHA-L and HRP labeled trigemino-thalamic neurons were seen in the JX and also in the Vpdm. Then, electron microscopic (EM) study was done, injections of kainic acid into the Vodm and HRP into the contralateral thalamus was performed simultaneously. After EM embedding, the JX and Vpdm regions were selected, ultrathin sections were cut and observed with EM. In both areas, axo-somatic and axo-dendritic synapses were seen between degenerated boutons and HRP labeled somata or dendrites. Namely, the Vodm projecting terminals synapsed on trigemino-thalamic neurons in the JX and Vpdm. Anyway, axo-dendritic synapses was the main type of observed synapses. Thus, the present work demonstrated 1. the JX containing a group of trigemno-thalamic neurons was a target of special projections froin the Vodm; 2. The Vodm neurons projected to the contralateral thalamus through the relay of JX and Vpdm neurons.     

Physiological and morphological characteristics of periodontal mesencephalic trigeminal neurons in the cat--intra-axonal staining with HRP

Intra-axonal recording and horseradish peroxidase (HRP) injection techniques were employed to define the response properties of periodontal mechanoreceptive afferents originating from the trigeminal mesencephalic nucleus (Vmes) and their morphological characteristics. The periodontal Vmes neurons were classified into two types: slowly adapting (SA) and fast adapting (FA) types. The central terminals of 7 SA and 4 FA afferents were recovered for detailed analyses. The whole profile of SA and FA neurons were unipolar in shape and their cell bodies were located in the dorsomedial parts of the Vmes. The united (U) fiber traveled caudally from the soma to the dorsolateral aspect of the trigeminal motor nucleus (Vmo), where it split into the peripheral (P) and C fibers with a T- or Y-shaped appearance. The P fiber joined the trigeminal sensory or motor tract. The C fiber descended caudally within Probst's tract. All 3 stem fibers issued main collaterals. The main collaterals of all neurons examined formed terminal arbors in the supratrigeminal nucleus (Vsup) and all but two SA neurons projected to the intertrigeminal region (Vint), while the projections to other nuclei of the trigeminal motor nucleus (Vmo), juxtatrigeminal region (Vjux), main sensory nucleus (Vp) and oral nucleus (Vo.r) differed between SA and FA afferents and between neurons of the same type. The SA and FA neurons were classified into three and two subgroups, respectively. The major differences in central projections between the two types were that all the FA neurons projected to the Vp or Vo.r but none of SA type and this relation was reversed in the projection to the Vjux, and that more than half of SA neurons projected to Vmo but only one FA neuron to the Vmo. The Vmes neurons which sent their collaterals into the Vmo had the P fiber passing through the tract of the trigeminal motor nerve. The average size of somata and mean diameters of U fibers and main collaterals from C fiber were significantly larger in SA neurons than FA neurons. The average size of fiber varicosities became smaller in the following nuclei, Vmo, Vsup, Vp, Vint and Vo.r, but not significant between the two functional types. The functional role of the periodontal Vmes afferents to jaw reflexes was discussed particularly with respect to their central projection sites in the brainstem nuclei.     

Morphology of single mesencephalic trigeminal neurons innervating periodontal ligament of the cat

The morphology of single neurons in the trigeminal mesencephalic nucleus (Vmes) that innervate periodontal ligament was studied in cats by the method of intraaxonal injection of horseradish peroxidase (HRP). Two kinds of Vmes neurons were distinguished on the basis of differences in axon profile and its central projection. The first type of Vmes neurons was unipolar in shape and its axon was divided into united (U), peripheral (P), and central axons (C). The U axon traveled caudally within the Vmes from the soma to the dorsolateral aspect of trigeminal motor nucleus (Vmo), where it split into the P and C axons with a T-shaped appearance. The P axon joined the spinal trigeminal tract across the trigeminal principal nucleus and ran within the tract and sensory root to exit the brainstem. The C axon traveled caudally within Probst's tract. All 3 axons issued axon collaterals. Axon collaterals from the U, P and the proximal C axons sent their terminal branches into the supra (Vsup) and intertrigeminal regions (Vint). Most axon collaterals from the C axon sent their terminal branches into the juxtatrigeminal regions (Vjuxta). The second type of Vmes neurons was bipolar and issued P and C axons. The C axon ran a short distance in the Vmes to leave the Vmes, and then it traveled caudolaterally in the rostrodorsomedial aspect of the Vmo. Finally, it entered in the Vmo and traveled caudally in the dorsolateral subdivision of the nucleus to its rostrocaudal mid-level. The C axon gave off massive axon collaterals.(ABSTRACT TRUNCATED AT 250 WORDS)     

Light and electron microscopic observations of a direct projection from mesencephalic trigeminal nucleus neurons to hypoglossal motoneurons in the rat

A direct projection from rat mesencephalic trigeminal nucleus (Vme) neurons to the hypoglossal nucleus (XII) motoneurons was studied using a double labeling method of anterogradely biotinylated dextran amine (BDA) tracing combined with retrogradely horseradish peroxidase (HRP) transport at both light and electron microscopic levels. BDA was iontophoresed unilaterally into the caudal Vme, and 7 days later HRP was injected into the ipsilateral tongue to label hypoglossal motoneurons. The BDA-labeled fibers were seen descended along Probst' tract and were traced to the caudal medulla. In this course, the fibers gave off axon collaterals bearing varicosities in the trigeminal motor nucleus (Vmo), the parvicellular reticular formation (PCRt), the dorsomedial portions of the subnuclei of oralis (Vodm) and interpolaris (Vidm) and in the XII ipsilaterally. The labeling of terminals was most dense in the PCRt at the levels of caudal pons and rostral medulla, which displayed a "dumbbell-shaped" form in the transverse planes. In the XII, labeled terminals were distributed mainly in the dorsal compartment of the nucleus. One hundred sixty-eight appositions made by BDA-labeled terminals on HRP-labeled motoneurons were seen in the dorsal compartment (71%) and in the lateral subcompartment (24%) of the ventral XII. Under electron microscopy BDA-labeled boutons containing clear, spherical synaptic vesicles were found to form synaptic contacts with the somata and dendrites of hypoglossal motoneurons with asymmetric specializations. The present study provides new evidence that the trigeminal proprioceptive afferent neurons terminate in the XII and make synaptic contacts with their motoneurons.     

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. © 2008 Wiley‐Liss, Inc.     

大鼠5—HT能纤维与三叉神经本体觉中枢四级通路Parvalbumin样阳性二级神经元接触

采用免疫荧光三重标记技术，对大鼠三叉神经本体觉中枢四级通路Parvalbumin（PV）样阳性二级神经元与5-羟色胺（5-HT）样阳性纤维间的联系在激光扫描共聚焦显微镜下进行了观察。结果表明：将葡聚糖胺结合的四甲基罗达明（TMR-DA）注入三叉神经感觉主核背内侧部（Vpdm）并对PV，5-HT及TMR-DA进行免疫荧光染色后，在三叉神经脊束核吻侧亚核背内侧部及其邻接的外侧网状结构（Vodm-LRF）中观察到5-HT样阳性纤维和PV/TMR-DA双标细胞；5-HT样阳性纤维的膨体或终扣与PV/TMR-DA双标细胞的胞体或树突有紧密接触。提示5-HT能终末与大鼠三叉神经本体觉中枢四级通路PV样阳性二级神经元间可能存在突触联系，并在二级神经元水平对三叉神经本体觉信息有调节作用。     

Morphology of single mesencephalic trigeminal neurons innervating masseter muscle of the cat

The morphology of functionally identified single axons of mesencephalic trigeminal neurons was studied in the cat by the method of intra-axonal injection of horseradish peroxidase (HRP). Each axon can be divided into united (U), peripheral (P) and central branches (C). The united axon (U) descends from its soma within the tract of the trigeminal mesencephalic nucleus to the dorsal aspect of the trigeminal motor nucleus (Vmo), where it splits into peripheral and descending central branches with a Y-shaped bifurcation. The peripheral axon (P) joins the motor root of the trigeminal nerve to exit the brainstem. The central axon (C) travels caudally within the juxtatrigeminal regions (or lateral reticular formation). All 3 branches issue axon collaterals that distribute terminal boutons within the dorsolateral subdivision of Vmo, supra- and intertrigeminal regions. Collaterals emanating from the central axon (C) except for its proximal segment travel ventrolaterally within the juxtatrigeminal regions, and send their terminal branches into the lateral boundaries adjacent to the spinal trigeminal nucleus. The trajectory of terminal branches distinguishes group Ia afferents from the possible group II afferents. The majority of terminal boutons are found to distribute in the supra- and intertrigeminal regions for group II afferent fibers and in the dorsolateral subdivision of Vmo for group Ia afferents.     

Neuronal circuitry and synaptic organization of trigeminal proprioceptive afferents mediating tongue movement and jaw-tongue coordination via hypoglossal premotor neurons

The neural framework and synaptic organization of trigeminal proprioceptive afferent-mediated jaw-tongue coordination were studied in rats using multiple electrophysiological and neuroanatomical approaches. Electrostimulation of the masseter nerve evoked short-latency responses (5.86 +/- 2.59 ms) in hypoglossal premotor pools including the parvocellular (PCRt) and intermediate (IRt) reticular nuclei and the dorsomedial part of the spinal trigeminal nucleus oralis (Vodm) and interpolaris (Vidm). Biocytin-labelled axon terminals from these areas traveled into the hypoglossal nucleus (XII) and contacted motoneurons. Double labelling of biotinylated dextran amine (BDA) tracing and cholera toxin B (CTB) transport demonstrated that labelled axons and terminals from the mesencephalic trigeminal nucleus (Vme) overlapped with XII premotor neurons in the alpha division and in PCRt, IRt, Vodm and Vidm. Confocal microscopic observations revealed that Vme terminals closely contacted XII premotor neurons. Dual labelling of intracellular neurobiotin staining of jaw-muscle spindle afferents (JMSAs) combined with horseradish peroxidase (HRP) retrograde transport revealed that 498 JMSA boutons apposed to 146 HRP-labelled premotor neurons. Electron microscopic observations demonstrated that 127 JMSA boutons made both axodendritic (68%) and axosomatic (32%) synapses with XII premotor neurons. Eighty-three per cent of synapses were asymmetric and the rest (17%) were symmetric. Thirty-nine per cent of JMSA boutons received presynaptic contacts from P-type terminals. Varieties of synaptic organizations were found. These results provide evidence that trigeminal proprioceptive afferents mediate jaw-tongue coordination through XII premotor neurons. Ultrastructural findings demonstrated that synapses between JMSA boutons and XII premotor neurons are predominantly excitatory, and synaptic transmission to XII motoneurons is modified on XII premotor neurons by presynaptic mechanisms. These frameworks and synaptic organizations are most probably the neural substrate for trigeminal proprioceptive afferent-mediated jaw-tongue coordination.     

Corticofugal projections to trigeminal motoneurons innervating antagonistic jaw muscles in rats as demonstrated by anterograde and retrograde tract tracing

Little is known about the organization of corticofugal projections controlling antagonistic jaw muscles. To address this issue, we employed retrograde (Fluorogold; FG) and anterograde (biotinylated dextran amine; BDA) tracing techniques in rats. Three groups of premotoneurons were identified by injecting FG into the jaw-closing (JC) and -opening (JO) subdivisions of the trigeminal motor nucleus (Vmo). These were 1) the intertrigeminal region (Vint) and principal trigeminal sensory nucleus for JC nucleus; 2) the reticular region medial to JO nucleus (RmJO) for JO nucleus; and 3) the parabrachial (Pb) and supratrigeminal (Vsup) nuclei, reticular regions medial and ventral to JC nucleus, rostrodorsomedial oralis (Vor), and juxtatrigeminal region (Vjuxt) containing a mixture of premotoneurons to both the nuclei. Subsequently, FG was injected into the representative premotoneuron structures. The JC and JO premotoneurons received main afferents from the lateral and medial agranular fields of motor cortex (Agl and Agm), respectively, whereas afferents to the nuclei with both JC and JO premotoneurons arose from Agl also and from primary somatosensory cortex (S1). Finally, BDA was injected into each of the three cortical areas representing the premotoneuron structures to complement the FG data. The Agl and Agm projected to reticular regions around the Vmo, whereas the Pb, Vsup, Vor, and Vjuxt received input from Agl. The S1 projected to the trigeminal sensory nuclei as well as to the Pb, Vsup, and Vjuxt. These results suggest that corticofugal projections to Vmo via premotoneuron structures consist of multiple pathways, which influence distinct patterns of jaw movements. J. Comp. Neurol. 514:368–386, 2009. © 2009 Wiley-Liss, Inc.     

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.     

Two types of jaw-muscle spindle afferents in the cat as demonstrated by intra-axonal staining with HRP

Intra-axonal records and horseradish peroxidase (HRP) injection techniques were employed to define the response properties of the jaw-closing muscle spindle afferents in the trigeminal mesencephalic nucleus (Vmes) and their morphological characteristics. The axonal trajectories of 9 spindle afferents from the masseter and 4 afferents from the temporalis were recovered for detailed analyses. Of 13 afferents, 6 cell bodies were stained and they were located at the rostrocaudal mid-levels of the Vmes. The central courses of the stem fibers were organized in a similar manner to the Vmes periodontal afferent nerves with the exception that peripheral (P) fibers of all spindle afferents passed through the trigeminal motor tract and root. On the basis of collateral terminal arborizations, the Vmes spindle afferents could be classified into two types: type I (n = 6) and type II (n = 7). Type I afferents sent their collaterals into the trigeminal motor nucleus (Vmo), intertrigeminal region (Vint) and juxtatrigeminal region (Vjux), but collaterals from the two neurons also projected to Vmes and the nucleus oralis (Vo). The collaterals from type II afferents formed their terminal arbors in the supratrigeminal nucleus (Vsup) in addition to the Vmo, Vint and Vjux, but collaterals from one neuron also projected to the Vo. In type I afferents, terminal arbors encompassed the whole Vmo including jaw-closing motoneurons. In contrast, boutons from type II afferents were restricted to a few small portions within the Vmo in proximity to its lateral and dorsal boundaries. The diameters of the united (U), central (C) and peripheral (P), fibers were larger in type I than type II afferents; those of the U fibers were statistically significant. Any differences between the two distinct types were not found in the response pattern to the sustained jaw opening. These results suggest that the difference of primary and secondary muscle-spindle afferent nerves is reflected in a distinctive morphology in the terminal arborizations and in the diameters of united fibers rather than the response patterns in deeply anesthetized cats.     

Morphological and functional properties of trigeminal nucleus oralis neurons projecting to the trigeminal motor nucleus of the cat

Horseradish peroxidase (HRP) was injected into the somata located in the rostrodorsomedial part (Vo.r) of the trigeminal nucleus oralis; an axonal projection to the trigeminal motor nucleus (Vmo) was demonstrated in two Vo.r neurons. The two neurons differed in their morphological and functional properties. The first Vo.r neuron responded to stimulation of low-threshold mechanoreceptors and its stem axon gave off massive axon collaterals that issued terminal branches to the dorsolateral subdivision of Vmo, Vo.r, and the medial and lateral parts of the lower brainstem reticular formation. The second Vo.r neuron was activated by stimulation of the tooth pulp or lingual nerve at twice longer latency than that of the first neuron. This stem axon was divided into two main ascending and one descending branches, and one of the main ascending branches was further bifurcated into two branches. The main non-bifurcated ascending branch gave off 4 collaterals, two of which sent terminal branches into the dorsolateral subdivision of Vmo and others into the Vo.r and juxta-trigeminal regions. The somato-dendroarchitectonic differences were also described in the two Vo.r neurons stained.     

Physiologic and morphologic properties of motoneurons and spindle afferents innervating the temporal muscle in the cat

Little is known about physiology and morphology of motoneurons and spindle afferents innervating the temporalis and on synaptic connections made between the two. The present study was aimed at investigating the above issues at the light microscopic level by using the intracellular recording and horseradish peroxidase or biotinamide labeling techniques and by the use of succinylcholine (SCh) for the classification of spindle afferents in the cat. Temporalis motoneurons had dendritic trees that ranged from a spherical form to an egg-shaped form. The shape deformation was more prominent for the dendritic trees made by motoneurons located closer to the nuclear border. No axon collaterals of the motoneurons were detected. On the basis of the values for the dynamic index after SCh infusion, temporalis spindle afferents were classified into two populations: presumptive groups Ia and II. The spindle afferents terminated mainly in the supratrigeminal nucleus (Vsup), region h, and the dorsolateral subdivision (Vmo.dl) of the trigeminal motor nucleus (Vmo). The proportion of group Ia afferent terminals was lower in the Vsup than that of group II afferents. In the Vmo.dl, the proportion of group Ia afferent terminals was nearly even throughout the nucleus, but that of group II afferent terminals increased in the more outlying regions. The proportion of terminal distribution in the central region of Vmo.dl was higher for group Ia than group II. The frequency of contacts (presumptive synapses) made by a single spindle afferent on a motoneuron was higher for group Ia than group II. The present study provided evidence that the central organization of spindle afferent neurons is different between groups Ia and II.     

Electron microscopic observation of synaptic connections of jaw-muscle spindle and periodontal afferent terminals in the trigeminal motor and supratrigeminal nuclei in the cat

Previous studies indicate that the trigeminal motor nucleus (Vmo) and supratrigeminal nucleus (Vsup) receive direct projections from muscle spindle (MS) and periodontal ligament (PL) afferents. The aim of the present study is to examine the ultrastructural characteristics of the two kinds of afferent in both nuclei using the intracellular horseradish peroxidase (HRP) injection technique in the cat. Our observations are based on complete or near-complete reconstructions of 288 MS (six fibers) and 69 PL (eight fibers) afferent boutons in Vmo, and of 93 MS (four fibers) and 188 PL (four fibers) afferent boutons in Vsup. All the labeled boutons contained spherical synaptic vesicles and were presynaptic to neuronal elements, and some were postsynaptic to axon terminals containing pleomorphic, synaptic vesicles (P-endings). In Vmo neuropil, MS afferent boutons were distributed widely from soma to distal dendrites, but PL afferent boutons predominated on distal dendrites. Most MS afferent boutons (87%) formed synaptic specialization(s) with one postsynaptic target while some (13%) contacting two or three dendritic profiles; PL afferents had a higher number of boutons (43%) contacting two or more dendritic profiles. A small but significant number of MS afferent boutons (12%) received contacts from P-endings, but PL afferent boutons (36%) received three times as many contacts from P-endings as MS afferents. In Vsup neuropil, most MS (72%) and PL (87%) afferent boutons formed two contacts presynaptic to one dendrite and postsynaptic to one P-ending, and their participation in synaptic triads was much more frequent than in Vmo neuropil. The present study indicates that MS and PL afferent terminals have a distinct characteristic in synaptic arrangements in Vmo and Vsup and provides evidence that the synaptic organization of primary afferents differs between the neuropils containing motoneurons and their interneurons. © 1996 Wiley-Liss, Inc.     

Central terminals of orofacial primary afferents and NADPH-diaphorase activity in the trigemino–solitary complex of rats

Nicotinamide adenine dinucleotide phosphate-diaphorase (NADPH-d) activity and the central terminal fields of branches of the mandibular and chorda tympani nerves were visualized histochemically at the same time using transganglionic transport of wheat germ agglutinin conjugated with horseradish peroxidase. The blue NADPH-d-positive neurons comprised a sparse network in the dorsomedial spinal trigeminal subnucleus oralis and a dense one in the rostral lateral division of the nucleus of the solitary tract. In the subnucleus caudalis, most labeled neurons were in the superficial zone, and smaller numbers were in the magnocellular zone. The NADPH-d-positive neurons in the subnucleus oralis and the nucleus of the solitary tract overlapped mostly with the transganglionically labeled terminal field from the lingual nerve, partly with the terminal field from the inferior alveolar and chorda tympani nerves, and rarely with the terminal field from the mental nerve. The NADPH-d-positive neurons in the dorsomedial paratrigeminal nucleus and subnucleus caudalis overlapped mostly with the terminal field from the lingual nerve, partly with the terminal field from the inferior alveolar and mental nerves and never with the terminal field from the chorda tympani. A statistically significant reduction in the number of NADPH-d-positive neurons was seen bilaterally in subnucleus oralis and the nucleus of the solitary tract when the lingual nerve was transected. Inflammatory insults to the lingual nerve or tooth pulps significantly increased the number of NADPH-d-positive neurons in subnucleus oralis, the nucleus of the solitary tract, and subnucleus caudalis. These results show that the NO/cyclic GMP system in the trigeminal and solitary nuclei is differentially regulated trans-synaptically by trigeminal afferents depending on the nucleus and sensory modality.     

An HRP study of the central projections of primary trigeminal neurons which innervate tooth pulps in the cat

Trigeminal ganglia and brain stem of adult cats were studied following HRP injections into tooth pulps or after exposure of the cut end of the inferior alveolar nerve to HRP. Ipsilateral ganglion cells within a wide range of sizes were labeled in both experimental situations, whereas no labeled cells were observed in the contralateral ganglion in any animal. Labeled central branches of tooth pulp and inferior alveolar neurons were observed in all subdivisions of the ipsilateral trigeminal sensory complex. Terminal labeling in the tooth pulp experiments was confined to the dorsomedial parts of the main sensory nucleus and subnuclei oralis and interpolaris. Caudal to the obex terminal labeling was restricted to the medial halves of laminae I, IIa and V of the medullary dorsal horn. In the inferior alveolar nerve experiments dense terminal labeling was observed in the dorsal parts of the main sensory nucleus and subnuclei oralis and interpolaris. Caudal to the obex terminal labeling was located throughout laminae I to V in contrast to the tooth pulp experiments. Neither of the two experimental situations offers any evidence for a bilateral or contralateral brain stem projection of primary trigeminal neurons.