Parvicellular Reticular Formation Research Articles

An origin of carotid vasodilation extends along the full extent of the parasympathetic parvicellular reticular region in the rat brainstem.

During grooming in rats, cranial parasympathetic activation leads to increased carotid artery blood flow, but the brainstem origin of this vasodilation signal is poorly understood. The aim of this study was to map brainstem sites wherein chemical stimulation with l-cysteine, an ionotropic excitatory amino acid receptor activator, can trigger carotid vasodilation in anesthetized intact and superior cervical sympathectomized (SCD) rats. The right side of the brainstem was accessed ventrally; arterial blood pressure and right carotid artery flow resistance were monitored. Employing microinjections of l-cysteine into the ipsilateral brainstem, vasodilation triggering sites were identified dorsal and rostral to the rostral ventrolateral medulla (RVLM) presympathetic pressor area, forming a parasagittal plate-like response zone that varied from rat to rat. In SCD rats (N=6), we observed a vasodilation zone in a region that included the RVLM pressor area, suggesting a parasympathetic origin. A similar zone was identified in intact rats (N=8), except that it did not include part of the RVLM pressor area. Dye injected into vasodilator sites labelled the rostral part of the parvicellular reticular formation (PcRt), consistent with a cranial parasympathetic origin. Employing mapping with L-cysteine, a putative neuromodulator, this study provides the first functional demonstration of a carotid vasodilation area that extends along the full extent of the parasympathetic rostral PcRt region in rats.

A Functionally and Anatomically Bipartite Vocal Pattern Generator in the Rat Brain Stem.

SummaryThe mammalian vocal pattern generator is situated in the brainstem but its exact structure is debated. We mapped these circuits in rats by cooling and microstimulation. Local cooling disrupted call production above an anterior and a posterior brainstem position. Anterior cooling affected predominantly high-frequency calls, whereas posterior cooling affected low-frequency calls. Electrical microstimulation of the anterior part led to modulated high-frequency calls, whereas microstimulation of the posterior part led to flat, low-frequency calls. At intermediate positions cooling did not affect calls and stimulation did not elicit calls. The anterior region corresponds to a subsection of the parvicellular reticular formation that we term the vocalization parvicellular reticular formation (VoPaRt). The posterior vocalization sites coincide with the nucleus retroambiguus (NRA). VoPaRt and NRA neurons were very small and the VoPaRt was highly myelinated, suggestive of high-speed processing. Our data suggest an anatomically and functionally bipartite vocal pattern generator.

Control Pathways from the Subcortical Limbic Structures to the Trigeminal Motor System in the Lower Brainstem: A Hodological Review

The organization of the emotion-related somatic motor behavior, including jaw movements, is governed not only by the cortical limbic system but also by the subcortical limbic system including the amygdala and hypothalamus. GABAergic neurons in the central amygdaloid nucleus (CeA) and glutamatergic neurons in the posterior lateral hypothalamus (PLH) exert inhibitory and excitatory influences, respectively, upon premotor neurons for the motor trigeminal nucleus (Vm) in the parvicellular reticular formation (RFp) of the medulla oblongata. The CeA also has an inhibitory influence on non-dopaminergic (probably GABAergic) neurons in the retrorubral field of the midbrain that send their axons to the RFp. Furthermore, the CeA and lateral hypothalamus including the PLH may also modulate Vm neurons via projections to the mesencephalic trigeminal nucleus which contains the cell bodies of primary afferent neurons conveying inputs from the masticatory muscle spindles or the periodontal ligament receptors to jaw closing motoneurons within the Vm. These pathways from the subcortical limbic structures to the trigeminal motor system in the lower brainstem underlie the regulation of emotional jaw movements.

The rostral parvicellular reticular formation neurons mediate lingual nerve input to the rostral ventrolateral medulla

In rats that had been anesthetized by urethane-chloralose, we investigated whether neurons in the rostral part of the parvicellular reticular formation (rRFp) mediate lingual nerve input to the rostral ventrolateral medulla (RVLM), which is involved in somato-visceral sensory integration and in controlling the cardiovascular system. We determined the effect of the lingual nerve stimulation on activity of the rRFp neurons that were activated antidromically by stimulation of the RVLM. Stimulation of the lingual trigeminal afferent gave rise to excitatory effects (10/26, 39%), inhibitory effects (6/26, 22%) and no effect (10/26, 39%) on the RVLM-projecting rRFp neurons. About two-thirds of RVLM-projecting rRFp neurons exhibited spontaneous activity; the remaining one-third did not. A half (13/26) of RVLM-projecting rRFp neurons exhibited a pulse-related activity, suggesting that they receive a variety of peripheral and CNS inputs involved in cardiovascular function. We conclude that the lingual trigeminal input exerts excitatory and/or inhibitory effects on a majority (61%) of the RVLM-projecting rRFp neurons, and their neuronal activity may be involved in the cardiovascular responses accompanied by the defense reaction.

Neurochemical Properties of the Synapses in the Pathways of Orofacial Nociceptive Reflexes

The brainstem premotor neurons of the facial nucleus (VII) and hypoglossal (XII) nucleus can integrate orofacial nociceptive input from the caudal spinal trigeminal nucleus (Vc) and coordinate orofacial nociceptive reflex (ONR) responses. However, the synaptoarchitectures of the ONR pathways are still unknown. In the current study, we examined the distribution of GABAergic premotor neurons in the brainstem local ONR pathways, their connections with the Vc projections joining the brainstem ONR pathways and the neurochemical properties of these connections. Retrograde tracer fluoro-gold (FG) was injected into the VII or XII, and anterograde tracer biotinylated dextran amine (BDA) was injected into the Vc. Immunofluorescence histochemical labeling for inhibitory/excitatory neurotransmitters combined with BDA/FG tracing showed that GABAergic premotor neurons were mainly distributed bilaterally in the ponto-medullary reticular formation with an ipsilateral dominance. Some GABAergic premotor neurons made close appositions to the BDA-labeled fibers coming from the Vc, and these appostions were mainly distributed in the parvicellular reticular formation (PCRt), dorsal medullary reticular formation (MdD), and supratrigeminal nucleus (Vsup). We further examined the synaptic relationships between the Vc projecting fibers and premotor neurons in the VII or XII under the confocal laser-scanning microscope and electron microscope, and found that the BDA-labeled axonal terminals that made asymmetric synapses on premotor neurons showed vesicular glutamate transporter 2 (VGluT2) like immunoreactivity. These results indicate that the GABAergic premotor neurons receive excitatory neurotransmission from the Vc and may contribute to modulating the generation of the tonic ONR.

Nociceptive Afferents to the Premotor Neurons That Send Axons Simultaneously to the Facial and Hypoglossal Motoneurons by Means of Axon Collaterals

It is well known that the brainstem premotor neurons of the facial nucleus and hypoglossal nucleus coordinate orofacial nociceptive reflex (ONR) responses. However, whether the brainstem PNs receive the nociceptive projection directly from the caudal spinal trigeminal nucleus is still kept unclear. Our present study focuses on the distribution of premotor neurons in the ONR pathways of rats and the collateral projection of the premotor neurons which are involved in the brainstem local pathways of the orofacial nociceptive reflexes of rat. Retrograde tracer Fluoro-gold (FG) or FG/tetramethylrhodamine-dextran amine (TMR-DA) were injected into the VII or/and XII, and anterograde tracer biotinylated dextran amine (BDA) was injected into the caudal spinal trigeminal nucleus (Vc). The tracing studies indicated that FG-labeled neurons receiving BDA-labeled fibers from the Vc were mainly distributed bilaterally in the parvicellular reticular formation (PCRt), dorsal and ventral medullary reticular formation (MdD, MdV), supratrigeminal nucleus (Vsup) and parabrachial nucleus (PBN) with an ipsilateral dominance. Some FG/TMR-DA double-labeled premotor neurons, which were observed bilaterally in the PCRt, MdD, dorsal part of the MdV, peri-motor nucleus regions, contacted with BDA-labeled axonal terminals and expressed c-fos protein-like immunoreactivity which induced by subcutaneous injection of formalin into the lip. After retrograde tracer wheat germ agglutinated horseradish peroxidase (WGA-HRP) was injected into VII or XII and BDA into Vc, electron microscopic study revealed that some BDA-labeled axonal terminals made mainly asymmetric synapses on the dendritic and somatic profiles of WGA-HRP-labeled premotor neurons. These data indicate that some premotor neurons could integrate the orofacial nociceptive input from the Vc and transfer these signals simultaneously to different brainstem motonuclei by axonal collaterals.

Amygdala connections with jaw, tongue and laryngo-pharyngeal premotor neurons

As the central nucleus (CE) is the only amygdaloid nucleus to send axons to the pons and medulla, it is thought to be involved in the expression of conditioned responses by accessing hindbrain circuitry generating stereotypic responses to aversive stimuli. Responses to aversive oral stimuli include gaping and tongue protrusion generated by central pattern generators and other premotor neurons in the ponto-medullary reticular formation. We investigated central nucleus connections with the reticular formation by identifying premotor reticular formation neurons through the retrograde trans-synaptic transport of pseudorabies virus (PRV) inoculated into masseter, genioglossus, thyroarytenoid or inferior constrictor muscles in combination with anterograde labeling of CE axons with biotinylated dextran amine. Three dimensional mapping of PRV infected premotor neurons revealed specific clusters of these neurons associated with different oro-laryngo-pharyngeal muscles, particularly in the parvicellular reticular formation. CE axon terminals were concentrated in certain parvicellular clusters but overall putative contacts were identified with premotor neurons associated with all four oro-laryngo-pharyngeal muscles investigated. We also mapped the retrograde trans-synaptic spread of PRV through the various nuclei of the amygdaloid complex. Medial CE was the first amygdala structure infected (4 days post-inoculation) with trans-synaptic spread to the lateral CE and the caudomedial parvicellular basolateral nucleus by day 5 post-inoculation. Infected neurons were only very rarely found in the lateral capsular CE and the lateral nucleus and then at only the latest time points. The data demonstrate that the CE is directly connected with clusters of reticular premotor neurons that may represent complex pattern generators and/or switching elements for the generation of stereotypic oral and laryngo-pharyngeal movements during aversive oral stimulation. Serial connections through the amygdaloid complex linked with the oro-laryngo-pharyngeal musculature appear quite distinct from those believed to sub-serve fear responses, suggesting there are distinct “channels” for the acquisition and expression of particular conditioned behaviors.

Central amygdaloid axon terminals are in contact with retrorubral field neurons that project to the parvicellular reticular formation of the medulla oblongata in the rat

The retrorubral field (RRF) contains numerous dopaminergic neurons and projects to the parvicellular reticular formation (RFp) of the medullary and pontomedullary brainstem, where many premotor neurons project to the orofacial motor nuclei. To know how the amygdala affects the RRF–RFp pathway in the rat, we first examined the synaptic organization between the central amygdaloid nucleus (CeA) fibers and the RFp-projecting RRF neurons by using combined anterograde and retrograde tracing techniques. After ipsilateral injections of biotinylated dextran amine (BDA) into the CeA and Fluoro-gold (FG) into the RFp, the prominent overlapping distribution of BDA-labeled axon terminals and FG-labeled neurons was found in the lateral part of the RRF ipsilateral to the injection sites, where the BDA-labeled axon terminals made symmetrical synapses with somata and dendrites of the FG-labeled neurons. Using a combination of retrograde tracing and immunohistochemistry for tyrosine hydroxylase (TH), we secondly demonstrated that the RFp-projecting RRF neurons were immunonegative for TH. Using a combination of anterograde tracing and immunohistochemistry for glutamic acid decarboxylase (GAD), we finally revealed that the CeA axon terminals in the RRF were immunoreactive for GAD. The present results suggest that GABAergic CeA neurons may exert inhibitory influences on non-dopaminergic RRF neurons that project to the RFp in the control of orofacial movements closely related to emotional behavior.

Multiple forebrain systems converge on motor neurons innervating the thyroarytenoid muscle

The present study investigated the central connections of motor neurons innervating the thyroarytenoid laryngeal muscle that is active in swallowing, respiration and vocalization. In both intact and sympathectomized rats, the pseudorabies virus (PRV) was inoculated into the muscle. After initial infection of laryngomotor neurons in the ipsilateral loose division of the nucleus ambiguus (NA) by 3 days post-inoculation, PRV spread to the ipsilateral compact portion of the NA, the central and intermediate divisions of the nucleus tractus solitarii, the Botzinger complex, and the parvicellular reticular formation by 4 days. Infection was subsequently expanded to include the ipsilateral granular and dysgranular parietal insular cortex, the ipsilateral medial division of the central nucleus of the amygdala, the lateral, paraventricular, ventrolateral and medial preoptic nuclei of the hypothalamus (generally bilaterally), the lateral periaqueductal gray, the A7 and oral and caudal pontine nuclei. At the latest time points sampled post-inoculation (5 days), infected neurons were identified in the ipsilateral agranular insular cortex, the caudal parietal insular cortex, the anterior cingulate cortex, and the contralateral motor cortex. In the amygdala, infection had spread to the lateral central nucleus and the parvicellular portion of the basolateral nucleus. Hypothalamic infection was largely characterized by an increase in the number of infected cells in earlier infected regions though the posterior, dorsomedial, tuberomammillary and mammillary nuclei contained infected cells. Comparison with previous connectional data suggests PRV followed three interconnected systems originating in the forebrain; a bilateral system including the ventral anterior cingulate cortex, periaqueductal gray and ventral respiratory group; an ipsilateral system involving the parietal insular cortex, central nucleus of the amygdala and parvicellular reticular formation, and a minor contralateral system originating in motor cortex. Hypothalamic innervation involved several functionally specific nuclei. Overall, the data imply complex CNS control over the multi-functional thyroarytenoid muscle.

Posterior lateral hypothalamic axon terminals are in contact with trigeminal premotor neurons in the parvicellular reticular formation of the rat medulla oblongata

This study was performed to understand the anatomical substrates of hypothalamic modulation of jaw movements. After cholera toxin B subunit (CTb) injection into the parvicellular reticular formation (RFp) of the rat medulla oblongata, where many trigeminal premotor neurons have been known to exist, numerous CTb-labeled neurons were found in the posterior lateral hypothalamus (PLH) bilaterally with a clear-cut ipsilateral dominance. After ipsilateral injections of biotinylated dextran amine (BDA) into the PLH and CTb into the motor trigeminal nucleus (Vm), the prominent distribution of BDA-labeled axon terminals around CTb-labeled neurons was found in the RFp region just ventral to the nucleus of the solitary tract and medial to the spinal trigeminal nucleus ipsilateral to the injection sites. Within the neuropil of the RFp, BDA-labeled axon terminals made an asymmetrical synaptic contact predominantly with dendrites and additionally with somata of the RFp neurons, some of which were labeled with CTb. It was further revealed that these BDA-labeled axon terminals were immunoreactive for vesicular glutamate transporter 2. The present data suggest that the PLH plays an important role in the control of jaw movements by exerting its glutamatergic excitatory action upon RFp neurons presynaptic to trigeminal motoneurons.

Lingual nerve modulation of the cardiac related activity in neurons of the rostral parvicellular reticular formation

Lingual nerve modulation of the cardiac related activity in neurons of the rostral parvicellular reticular formation

The effect of lingual nerve stimulation on the cardiac-related activity of the parvicellular reticular formation neurons, and their projections to the rostral ventrolateral medulla

The effect of lingual nerve stimulation on the cardiac-related activity of the parvicellular reticular formation neurons, and their projections to the rostral ventrolateral medulla

Rostral parvicellular reticular formation neurons projecting to rostral ventrolateral medulla receive cardiac inputs in anesthetized rats

The rostral parvicellular reticular formation (rRFp) was explored electrophysiologically in urethane-chloralose anesthetized rats. Spontaneously-active neurons that exhibited a pulse-related activity were recorded and tested for their projections to the rostral ventrolateral medulla (RVLM). About one-third (10/29) of the rRFp neurons that exhibited a pulse-related activity were antidromically activated by RVLM stimulation with conduction velocities between 0.2–4.4 m/s and fell within the B and C fibre range. A majority (8/10) of these neurons had a low (<10 spikes/s) mean firing rate, whereas a small proportion (2/10) had a high (>15 spikes/s) mean firing rate. These findings suggest a direct pathway from the rRFp to the RVLM and suggest that neurons projecting to the RVLM receive cardiac inputs and can modulate RVLM neuronal activity.

Role of the parvicellular reticular formation in facilitating the jaw-opening reflex in rats by stimulation of the red nucleus

In a previous study, we have shown that electrical and chemical stimulation of the red nucleus (RN) facilitates the jaw-opening reflex (JOR). The RN sends projection fibers bilaterally, with contralateral dominance, to the part of the parvicellular reticular formation (RFp) containing premotor neurons projecting to the trigeminal motor nucleus. This implies that RN-induced facilitation of the JOR might be mediated via last-order neurons in the RFp. Here, we address this issue by investigating whether microinjection of lidocaine or l-glutamate into the RFp affects RN-induced modulation of the JOR. Experiments were performed on rats anesthetized with urethane–chloralose. The JOR was evoked by electrical stimulation of the inferior alveolar (IA) nerve and was recorded as an electromyographic response from the anterior belly of the digastric muscle. Conditioning stimulation was delivered unilaterally to the RN 12 ms before the IA test stimulation. We found that local injections of 2% lidocaine (0.5 μl) into the RFp, contralateral to the RN, significantly ( P < 0.05) reduce RN-induced facilitation of the JOR, whereas corresponding injections of 0.1 mM l-glutamate (0.5 μl) significantly ( P < 0.05) increase it. These results suggest that the facilitatory effect of RN stimulation on the JOR is mediated partly by a relay in the RFp.

Role of GABA A receptors in the retrorubral field and ventral pallidum in rat jaw movements elicited by dopaminergic stimulation of the nucleus accumbens shell

The role of γ-aminobutyric acid (GABA) A receptors in the retrorubral field in the production of rat repetitive jaw movements was examined, as this nucleus receives a GABAergic, inhibitory input from the nucleus accumbens and is connected with the parvicellular reticular formation, a region that is directly connected with the orofacial motor nuclei. The GABA A receptor antagonist bicuculline (150 ng/0.2 μl per side) significantly produced repetitive jaw movements when injected bilaterally into the retrorubral field, but not the ventral pallidum. The effects of bicuculline were GABA A receptor specific, because the effects were abolished by muscimol, a GABA A receptor agonist, given into the same site. The bicuculline-induced jaw movements differed qualitatively from those elicited by injection of a mixture of (±)-6-chloro-7,8-dihydroxy-3-allyl-1-phenyl-2,3,4,5-tetrahydro-1 H-3-benzazepine-7,8-diol (SKF 82958; 5 μg) and quinpirole (10 μg), agonist at dopamine D1 and D2 receptors respectively, into the nucleus accumbens shell. Nevertheless, bilateral injections of muscimol (10 ng, 25 ng and 50 ng/0.2 μl per side) into the retrorubral field significantly inhibited jaw movements evoked by the dopamine D1/D2 receptor stimulation in the nucleus accumbens shell. Bilateral injections of bicuculline (50 ng and 150 ng/0.2 μl per side) also reduced the dopamine D1/D2 receptor-mediated jaw movements. Essentially similar effects were obtained when muscimol and bicuculline were given into the ventral pallidum, a region that is also known to receive GABAergic inhibitory inputs from the nucleus accumbens. In conclusion, GABA A receptor blockade in the retrorubral field elicits characteristic repetitive jaw movements, and the GABA A receptors in that region as well as in the ventral pallidum modulate the accumbens-specific, dopamine D1/D2 receptor-mediated jaw movements.

Amygdaloid axon terminals are in contact with trigeminal premotor neurons in the parvicellular reticular formation of the rat medulla oblongata

After ipsilateral injections of biotinylated dextran amine (BDA) into the central nucleus of the amygdala (ACe) and cholera toxin B subunit (CTb) into the motor trigeminal nucleus (Vm) in the rat, numerous BDA-labeled axons with bouton-like varicosities were distributed bilaterally with a clear-cut ipsilateral dominance in the parvicellular reticular formation (RFp), where many CTb-labeled neurons existed bilaterally with slightly ipsilateral dominance. The prominent overlapping distribution of these labeled axons and neurons was found in the RFp region just ventral to the nucleus of the solitary tract and medial to the spinal trigeminal nucleus throughout the caudalmost part of the pons and the rostral half of the medulla oblongata. Within the neuropil of the RFp region in the rostral half of the medulla oblongata, BDA-labeled axons made symmetrical synaptic contacts predominantly with the dendrites and additionally with the somata of RFp neurons, some of which were labeled with CTb. These data suggest that output signals from the ACe may be transmitted disynaptically to the Vm via the RFp neurons in the medulla oblongata for the control of jaw movements.

Neural activities in the substantia nigra modulated by stimulation of the orofacial motor cortex and rhythmical jaw movements in the rat

Neurons related to jaw movements in the substantia nigra pars reticulata were explored by examining changes in their neural activities in response to electrical stimulation of the orofacial sensorimotor cortex and during rhythmical jaw movements induced by mechanical stimulation applied to the oral cavity in the rat. Out of 80 neurons tested, 59 showed changes in their firing patterns of activities in response to the electrical stimulation of the cortex. The responding neurons were mainly located in the dorsolateral part of the substantia nigra pars reticulata. The substantia nigra pars reticulata neurons showing responses were classified into the following five types according to their response patterns: (1) an inhibition preceded by an early excitation and followed by a late excitation ( n=26), (2) an inhibition preceded by an early excitation but not followed by a late excitation ( n=7), (3) an inhibition not preceded by an early excitation but followed by a late excitation ( n=2), (4) an inhibition without early or late excitations ( n=7) and (5) an excitation without an inhibition ( n=17). Out of 18 neurons responding to the cortical stimulation, 11 (61.1%) increased or decreased their neural activities during rhythmical jaw movements. Some of these neurons had a projection to the lateral part of the superior colliculus ( n=5) and/or to the parvicellular reticular formation ( n=2). These results provide first neurophysiological evidence for neurons in the dorsolateral part of the substantia nigra pars reticulata with inputs from and outputs to the areas related to jaw movements. These neurons may participate in the control of jaw movements in the rat.

Rhombencephalic pathways and neurotransmitters controlling deglutition

Information from neuronal pathway tracing and pharmacologic microstimulation studies, in conjunction with electrophysiological data, has begun to coalesce into a coherent, if still incomplete, picture of the brain stem circuitry responsible for generating the motor patterns underlying deglutition and esophageal peristalsis in the rat. The intermediate, interstitial, and ventral subnuclei of the solitarius complex appear to play a pivotal part, as evidenced by their viscerosensory inputs and extensive projections to the parvicellular intermediate reticular formation of the medulla, to the brain stem deglutitive motor neuron pools, and to general visceral efferent preganglionic neurons controlling the upper alimentary tract striated and smooth musculature, respectively. The dense projections of the solitarial central subnucleus form a separate subcircuit controlling esophageal, and also some aspects of gastric, motility. Although not extensive, direct connections between the latter subnucleus and interneurons coordinating the buccopharyngeal stage of swallowing appear to exist. In both subcircuits, fast information transfer uses excitatory amino acidergic transmission by means of several glutamate-receptor subtypes. Release from tonic GABAergic inhibition exerted by local solitarial interneurons may provide a mechanism for triggering deglutitive premotoneuronal activity. Local or reticular cholinergic neurons are implicated in pharyngoesophageal coupling and the generation of propulsive esophagomotor output. The solitary interneurons under investigation engage in complex local dendritic and axonal projections within the solitarius complex. Further analysis of these local circuits and their transmitters should yield essential clues regarding the mechanisms underlying deglutitive motor pattern generation.

Projections from the red nucleus to the parvicellular reticular formation and the cervical spinal cord in the rat, with special reference to innervation by branching axons

After biotinylated dextranamine injection into the dorsal part of the red nucleus (RN) in the rat, labeled axons were distributed contralaterally in the lateral tegmental field including the parvicellular reticular formation (RFp), and ipsilaterally in the medial reticular formation. In the cervical spinal cord, labeled axons were present bilaterally with a contralateral dominance mainly in laminae V–VI and the dorsal part of laminae VII. After ipsilateral injections of rhodamine dextranamine, Fluoro-ruby (FR) into the RFp and Fluoro-gold (FG) into the upper cervical spinal cord, a population of FR-labeled neurons was found in the dorsal part of the contralateral RN, whereas the majority of FG-labeled neurons were located more ventrally. However, some of them were intermingled with FR-labeled neurons, and as many as one-third of FR-labeled neurons were labeled with FG. After combined injections of FR into the RFp and FG into the lower cervical spinal cord, RN neurons labeled with FG existed more ventrally than those retrogradely labeled from the upper cervical spinal cord, and less than 10% of FR-labeled neurons were labeled with FG. The present data suggest that axon collateral innervation of the RFp and the upper cervical spinal cord by single RN neurons may be responsible for coordinating head and orofacial movements.

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.