Subnucleus Interpolaris Research Articles

Physiological Identification of Jaw-Movement-Related Neurons in the Trigeminal Nucleus of Cats

Although neurons responsive to jaw movements have been identified in most parts of the trigeminal brainstem nuclei, little is known about how this information is relayed to the thalamus and ultimately to the cortex for kinesthetic functions and sensorimotor integration. The present extracellular recording experiments showed that a substantial amount of movement-related information is relayed to the thalamus through the caudal part of subnucleus interpolaris (Vi) in adult cats. Vertical jaw displacements, natural mechanical stimuli, and electrical stimulation of the masseter nerve were used to determine the receptive fields and response properties of movement-related neurons. Movement-related responses were observed in 161 units. The receptive fields of these units were located in the masseter muscle, other deep structures, hairy skin, oral mucosa, or some combination of these structures (i.e., convergent). The latency of units responding to masseter nerve stimulation ranged from 1.0 msec to 20 msec, which suggested that some movement-related information was provided by smaller-diameter muscle afferents. Movement responses were either tonic or phasic. Tonic units showed continuous firing at some jaw position; some of these showed a "dynamic" response to jaw displacement. Phasic units were only active, or showed increased activity, when the jaw moved through a specific position. Seventy-one movement-related units were activated by stimulation from the contralateral ventroposteromedial nucleus (VPM) of the thalamus. Most of the brainstem recording sites were located in the dorsal part of Vi between the caudal pole of the facial motor nucleus and the obex. Neurons in caudal Vi may be important for facial kinesthesia.(ABSTRACT TRUNCATED AT 250 WORDS)

Convergent projections of trigeminal afferents from the principal nucleus and subnucleus interpolaris upon rat ventral posteromedial thalamic neurons

Synaptic boutons originating from the trigeminal principal sensory nucleus (PrV) and the subnucleus interpolaris (SpI) which contact rat ventral posteromedial (VPM) neurons are similar in appearance. They are large boutons and contain a moderate packing density of round synaptic vesicles and establish asymmetric (Gray type I) synaptic contacts principally on dendrites and occasionally on somata. These boutons are similar to RL boutons of dorsal column nuclei and spinothalamic tract origin found in the ventral posterolateral thalamic nucleus. The boutons of PrV and SpI origin have an overlapping distribution on proximal dendrites of VPM neurons. Double labeling using degeneration and WGA-HRP shows that boutons from PrV and SpI contact the same VPM neuron confirming there is convergence of trigeminal afferents in VPM.

Neurons in the rat spinal trigeminal complex driven by corneal nociceptors: receptive-field properties and effects of noxious stimulation of the cornea

1. A survey of the receptive-field properties of neurons in the spinal trigeminal complex driven by stimulation of corneal afferents has been carried out. The afferent inputs to these neurons from the cornea and from the adjacent skin were studied as well as changes in the excitability of the cells and in the size of their receptive fields after thermal noxious stimulation of the cornea. 2. Single-unit electrophysiological recordings were made in pentobarbitone anesthetized rats from 54 neurons all of which were activated by mechanical stimulation of the ipsilateral cornea. Seventeen of these neurons were activated only by corneal stimulation and the other 37 had an additional cutaneous receptive field in the periorbital skin. Of the 37 neurons with a cutaneous receptive field, 29 were activated exclusively by noxious stimulation of the skin (Class 3) and the remaining 8 were driven by both innocuous and noxious cutaneous stimuli (Class 2). 3. All of the neurons were located in the ventro-lateral area of a region of the spinal trigeminal complex between +0.5 mm and -1.0 mm from the obex. This area corresponds to the most caudal part of subnucleus interpolaris, the transition zone between interpolaris and caudalis and the rostral half of subnucleus caudalis. Most neurons were located in the superficial layers of this part of the spinal trigeminal complex. No differences were observed between the locations of the recording sites of neurons with an exclusive corneal input and those with a corneal and a cutaneous receptive field.(ABSTRACT TRUNCATED AT 250 WORDS)

Morphology and topography of identified primary afferents in trigeminal subnuclei principalis and oralis.

1. Intra-axonal recording, receptive field mapping, horseradish peroxidase injection, cytochrome oxidase staining, and computer-assisted reconstruction/morphometric methods were used to elucidate the structure and topography of trigeminal primary afferent collaterals in the normal adult rat. Prior studies focused on trigeminal brain stem subnuclei interpolaris and caudalis. This work is extended here to the remaining 2 subnuclei, principalis (PrV) and oralis (SpVo), where collaterals from 66 axons in 37 adult rats were studied. In nine rats, three to five axons were stained for within-nucleus comparisons of different fibers. Quantitative analyses were restricted to vibrissa sensitive fibers. 2. All of the axons conducted rapidly with small, low-threshold receptive fields. The majority responded to vibrissa deflection (n = 47); the remainder responded to guard hair deflection; gentle pressure applied to hairy skin, glabrous skin, lingual mucosa, or an incisor; or jaw movement. All descended in the trigeminal sensory root where some bifurcated into ascending and descending branches. Each well-stained fiber gave rise to transversely oriented collaterals in PrV and SpVo. 3. Within PrV and SpVo, fibers with differing adaptation properties and receptive fields had indistinguishable collateral morphologies. Arbors from single axons were rostrocaudally discontinuous, small relative to collaterals in subnuclei interpolaris and caudalis, circumscribed and topographically organized in a manner consistent with cytochrome oxidase and bulk-labeled primary afferent staining patterns. In SpVo and caudal PrV, the map is inverted with the nose pointing medially. In rostral PrV, the map turns 90 degrees such that the nose points dorsally. 4. Axons had different quantitative properties along the rostrocaudal axis of the trigeminal brain stem complex. Whereas arbors subtended similar transverse areas throughout PrV and SpVo, collaterals in the rostral third of PrV had a relatively low bouton density. Arbors in the caudal two thirds of PrV had the highest bouton density. Arbors in SpVo tended to be more variable in size and shape than those of caudal PrV, and their bouton numbers were significantly lower than in PrV. 5. In PrV, arbors were largely confined to somatotopically corresponding cytochrome oxidase patches, precluding significant overlap of neighboring whisker projections. In SpVo, termination sites were not as strictly confined and numerous examples of within- and between-row overlap were obtained for whisker afferents in cases where multiple axons were stained.(ABSTRACT TRUNCATED AT 400 WORDS)

Microinjections of glutamate within trigeminal subnucleus interpolaris alters adrenal and autonomic function in the cat

The influence of rostral portions of the trigeminal sensory complex on adrenal and autonomic function was assessed by microinjections of l-glutamate (500 or 5 mM, 100 nl) directed at subnucleus interpolaris (Vi) or at the nucleus principalis/subnucleus oralis level (Vp/Vo) in chloralose-anesthetized cats. Microinjections of glutamate (500 mM) within Vi evoked prompt (by + 1 min) dose-related increases in the adrenal secretion of epinephrine ( +11.4±2.5ng/min, P < 0.001), adrenal blood flow ( +0.19±0.06 ml/min, P |s 0.05), mean arterial pressure ( +6.6±3.0mmHg, P < 0.025) and heart rate ( +8.0±2.7beats/min, P < 0.01, n = 16). Microinjections of lower doses of l-glutamate ( 5mM, n = 7) within Vi had no effect. Microinjections of 500 mM glutamate within Vp/Vo ( n = 15) or within the spinal trigeminal tract ( n = 13) had no consistent effect on adrenal or autonomic function. Plasma concentrations of ACTH were not altered significantly by glutamate regardless of dose or of the site of injection. The results suggest that local release of glutamate within Vi, but not within Vp/Vo, influences adrenal and autonomic function. Together with previous results obtained after injections of glutamate within subnucleus caudalis, these data indicate that glutaminergic input to both Vi and to more caudal portions of the spinal trigeminal nucleus contribute to the control of autonomic function such as that which often accompanies trigeminal nociception.

Structure-function relationships in rat brainstem subnucleus interpolaris. X. Mechanisms underlying enlarged spared whisker projections after infraorbital nerve injury at birth

Prior studies indicate that the central projections of noninfraorbital vibrissae occupy greater than normal transverse areas in the rat trigeminal brainstem complex after infraorbital nerve section at birth. Here, we assessed the development of this phenomenon and possible underlying mechanisms. Cytochrome oxidase patches representing spared supraorbital (SO) or posteroorbital (PO) whiskers in the trigeminal subnucleus interpolaris (SpVi) were not reliably larger than those on the control side 24 hr after the infraorbital lesion. By 72 hr, SO and PO patches were 91% and 28% larger than those on the control side. Reliable increases were also observed on postnatal day 5 (PND5), PND7, and PND10 for the SO (59%, 65%, 66%) and PO (23%, 44%, 51%) patches. To test the hypothesis that central reorganization reflects the maintenance of peripheral supernumerary axons, myelinated and unmyelinated axons in SO vibrissa follicles were counted at PND0, PND7, PND17, and PND60. A corollary hypothesis, that peripheral regeneration errors result in both SO and surviving infraorbital axons, contributing to central SO patches, was tested with retrograde double-labeling methods. Both hypotheses were rejected. Thus, enlargement of SO patches is not due to either the maintenance of an immature peripheral innervation pattern, or regeneration of infraorbital axons into SO follicles. To determine if the enlargement of SO and PO patches produced by infraorbital nerve section is due to an activity-dependent competitive disadvantage imposed upon infraorbital afferents, TTX or bupivicaine was applied to the intact infraorbital nerve over the first 5-9 postnatal days. Brainstem maps developed normally and SO and PO patch areas were unaffected. Thus, impulse activity-based mechanisms do not appear to contribute to injury-induced patch enlargement. To test the hypothesis that patch enlargement is due to central collateral reorganization, intra-axonal recording and staining methods were applied to control and spared-whisker primary afferents in adult rats. Total bouton or collateral numbers did not differ in SpVi; however, arbor areas were reliably larger in experimental (14,879 +/- 350 microns 2) versus control (5527 +/- 1811 microns 2) fibers.(ABSTRACT TRUNCATED AT 400 WORDS)

Galanin immunoreactivity reveals a vibrissae-related primary afferent pattern in perinatal rats after neonatal infraorbital nerve transection

Immunocytochemistry was used to demonstrate the distribution of galanin immunoreactivity in the trigeminal (V) brainstem complex of normal rats and rats that sustained neonatal transection of the infraorbital nerve (ION, the V branch that supplies the mystacial vibrissae). In normal adult and perinatal (postnatal day [P-] 7) rats, there is very little galanin immunoreactivity in the rostral part of the V brainstem complex. However, there is dense immunoreactivity for this peptide in layers I and II of V subnucleus caudalis (SpC). There was a marked upregulation of galanin immunoreactivity at all levels of the V brainstem complex of P-7 rats that sustained ION transection on P-0. In V subnucleus interpolaris and the magnocellular part of SpC, this immunoreactivity occurred in clusters which had a pattern resembling that of the mystacial vibrissae. In animals examined at 30 days or more after ION transection, there were a few more galanin-immunoreactive fibers on the deafferented side of the brainstem than on the intact side, but there was no evidence of a vibrissae-related pattern. Our interpretation of these results is that galanin immunocytochemistry reveals a vibrissae-related pattern in the central arbors of axotomized primary afferents for at least 1 week after they have been disconnected from the periphery.

Trigeminal nuclear complex of the ferret: anatomical and immunohistochemical studies.

In order to establish the ferret as an animal model for studies of trigeminal pain, we describe the cytoarchitecture and neurochemistry of the trigeminal nuclear complex in the ferret and compare them to those of the cat and rat. The complex was divided as previously described, but the ferret differed in the extent of the nuclear boundaries. The neuroanatomical istribution of substance P-, calcitonin gene-related peptide-, galanin-, enkephalin-, serotonin-, somatostatin-, neuropeptide Y-, and neurotensin-immunoreactivity was determined throughout the rostrocaudal extent of the complex. In subnucleus caudalis, substance P-, calcitonin gene-related peptide-, enkephalin-, serotonin-, somatostatin-, neuropeptide Y-, and galanin-immunoreactivity was densest in laminae I and II. In subnucleus interpolaris, immunoreactivity for all the above neurochemicals was most dense along the lateral border and the ventral third of the caudal part of the subnucleus. Enkephalin-immunoreactive cell bodies were present in subnucleus caudalis and interpolaris. In subnucleus oralis, labelling for substance P, calcitonin gene-related peptide, galanin, enkephalin, and serotonin was most prominent in the dorsomedial part of the subnucleus. Somatostatin-immunoreactive cell bodies were distributed throughout the spinal nucleus. Labelling of serotonin, substance P, calcitonin gene-related peptide, galanin, enkephalin, and somatostatin was present in the main sensory nucleus. The motor nucleus contained fibers immunoreactive for substance P, enkephalin, serotonin and neuropeptide Y, and cell bodies immunoreactive for calcitonin gene-related peptide. The majority of neurotensin-immunoreactivity was found at the level of subnucleus caudalis, where it was densest in the trigeminal extension of the lateral cervical nucleus. The distribution of peptides in this species throughout the spinal nucleus is consistent with the notion that all the subnuclei may be involved in the processing of nociceptive inputs.

Cat trigeminal neurons innervated from the planum nasale: their medullary location and their responses to mechanical stimulation.

Experiments were performed to determine whether the receptors of the glabrous skin of the cat planum nasale (PN) could function in tactile analysis. Trigeminal projection sites of the PN were first identified using transganglionic transport of wheat germ agglutinin-horseradish peroxidase and horseradish peroxidase. Restricted projection sites were identified in this way among the interstitial neurons of the trigeminal tract, in the dorsal horn of the medulla, in subnucleus interpolaris and to a lesser extent in subnucleus oralis. Electrophysiological recording in the trigeminal spinal nucleus confirmed the major neuroanatomical findings and confirmed the paucity of PN projections to the trigeminal system. Most neurons innervated from the PN have small receptive fields, are rapidly adapting and responsive to PN vibration at amplitudes as low as 10 micros. Neurons could be entrained at frequencies below 80 Hz. This upper limit for entrainment presumably reflects the lack of pacinian corpuscles in the PN. A limited number of slowly adapting neurons were found, but only responded to PN displacements of 400 microns and above. The data suggest that the PN can function in tactile analysis to a limited degree. The significance of these findings is considered with respect to the organization of neural systems controlling head movement.

Physiological properties and morphological characteristics of cutaneous and mucosal mechanical nociceptive neurons with A‐δ peripheral axons in the trigeminal ganglia of crotaline snakes

Primary A-delta nociceptive neurons in the trigeminal ganglia of immobilized crotaline snakes were examined by intrasomal recording and injection of horseradish peroxidase in vivo. Thirty-four neurons supplying the oral mucosa or facial skin were identified as A-delta nociceptive neurons which responded exclusively to noxious mechanical stimuli and had a peripheral conduction velocity ranging from 2.6 to 15.4 m/s. These neurons were subdivided into a fast-conducting type (FC-type) and a slowly conducting type (SC-type). Neurons of both types had a receptive field limited to a single spot which responded to pin prick stimulus with a threshold of more than 5 g. The FC-type neurons had a narrow spike followed by a shorter after-hyperpolarization. In contrast, SC-type neurons exhibited a broad spike with a hump on the falling phase and a longer after-hyperpolarization. The diameters of the stem, central and peripheral axons of the FC-type neurons were significantly thicker than those of the SC-type neurons, but there was no statistical difference in the soma size of the two types. Central axons of both types of neurons were thinner than their stem and peripheral axons. Dichotomizing fibers of peripheral axons were observed within the ganglion on 3 neurons. Central axons of the FC-type neurons terminated ipsilaterally in the nucleus principalis, the subnucleus oralis, interpolaris and caudalis and the interstitial nucleus, whereas those of the SC-type neurons generally projected only to the caudal half of the subnucleus interpolaris, subnucleus caudalis and interstitial nucleus ipsilaterally. The present data showed for the first time the physiological and morphological heterogeneity of the primary trigeminal A-delta nociceptive neurons and revealed that the trigeminal nucleus principalis and all the subdivisions of the trigeminal descending nucleus are involved in nociception as relay nuclei, but the subnucleus caudalis and the caudal half subnucleus interpolaris are the essential relay sites of the primary nociceptive afferents supplying the oral mucosa and facial skin. The interstitial nucleus also appears to play an important role in orofacial nociception.

Effects of neonatal transection of the infraorbital nerve upon the structural and functional organization of the ventral posteromedial nucleus in the rat.

The present study examined the way in which an indirect partial deafferentation of the medial portion of the ventrobasal complex (VPM/VPL) induced by neonatal transection of the infraorbital nerve (ION) altered the structural and functional properties of its constituent neurons. This manipulation significantly reduced the volume of the contralateral VPM/VPL. In addition, cell counts in Nissl-stained material revealed a significant reduction of the number of VPM/VPL neurons contralateral to neonatal ION transection. We also analyzed the effect of neonatal ION transection on the soma-dendritic morphology of individual neurons in the ventral posteromedial nucleus of the thalamus (VPM) by intracellular injection of horseradish peroxidase (HRP) in vivo and Lucifer yellow in fixed slices. Neonatal transection of the ION resulted in increased dendritic length, area, and volume of VPM neurons in both preparations; however only the changes observed in fixed slices reached statistical significance. Alterations in the functional characteristics of VPM neurons were also observed following neonatal nerve damage. There was a significant decrease in the percentage of vibrissae-sensitive neurons and a corresponding increase in the percentages of neurons responsive to guard hair deflection or that were unresponsive to peripheral stimulation. Neonatal nerve damage also resulted in significantly longer latencies of VPM cells after stimulation of either trigeminal nucleus principalis or subnucleus interpolaris. The present results indicate that the development of normal response properties and soma-dendritic morphology of VPM neurons is dependent upon intact afferent input during development. Indirect partial deafferentation of VPM/VPL by neonatal transection of the ION results in reduced neuron number, which may result in decreased competition among the dendrites of these neurons. This proposal is consistent with observations of increased dendritic dimensions of VPM neurons contralateral to neonatal ION damage.

Responses of trigeminal subnucleus interpolaris neurons to afferent inputs from deep oral structures

Responses of trigeminal subnucleus interpolaris neurons to natural and electrical stimulation of the temporomandibular joint (TMJ) and the masseter muscle (Mm) were studied in the anesthetized rat. Interpolaris neurons could be placed in one of three classes determined by their responses to noxious and innocuous stimuli: Class I, excited by only innocuous stimuli; Class II, excited by only noxious stimuli; Class III, excited by both. In each class, the conduction velocities of the primary afferents were roughly equivalent to those of small-diameter fibers. Approximately 70% of the interpolaris neurons tested received nociceptive inputs from the TMJ and/or the Mm. Most of these neurons had extensive convergence of afferent inputs, including the TMJ, the Mm, and/or the facial skin. The results suggest involvement of interpolaris neurons in the characteristics of deep pain, such as its spread and referral.

Normal development and effects of neonatal infraorbital nerve damage upon the innervation of the trigeminal brainstem complex by primary afferent fibers containing calcitonin gene‐related peptide

Immunocytochemistry was used to study the normal development and response to infraorbital nerve (ION) damage of the innervation of the trigeminal (V) brainstem complex by axons recognized by an antibody directed against calcitonin gene-related peptide (CGRP). CGRP-like immunoreactivity (CGRPLI) was present in axons that occupied the outer V spinal tract (TrV) at all levels of the V brainstem complex. Almost no fibers terminated within V nucleus principalis (PrV), but there was dense CGRPLI in the supratrigeminal nucleus. There was also very little CGRPLI within rostral V subnucleus oralis (SpO). However, in the caudal one-half of the nucleus, a dense elongated patch of immunoreactivity was consistently present just medial to TrV. Only occasional CGRP-positive axons could be seen within V subnucleus interpolaris (SpI), but the paratrigeminal nucleus contained dense immunoreactivity. Trigeminal subnucleus caudalis (SpC) also contained CGRPLI that was very dense in lamina I and the outer portion of lamina II. Scattered terminals were also present in layers III and IV and dense terminal clusters were in lamina V. CGRP-immunoreactive neurons were present in the V ganglion by embryonic (E-) day 16 and immunoreactive axons could be seen in the V brainstem complex on E-17. At birth, CGRP-positive axons in the V brainstem complex had achieved a distribution very similar to that in adult rats. The major difference between the patterns of labelling in neonates and adults was the presence of relatively large numbers of CGRP-positive fibers in ventral PrV and SpO of the former animals. The disappearance of these fibers was completed by the middle of the third postnatal week. Transection of the ION on the day of birth had little effect upon CGRP in SpO, SpI, and SpC, but it did result in an increase in CGRP-positive fibers in PrV ipsilateral to the damaged nerve. When considered together with previous findings, these results suggest that CGRP-positive axons express this peptide well after they have entered the V brainstem complex and that the central terminal field of these fibers is not substantially altered by a manipulation which results in the death of nearly 60% of all V primary afferent neurons.

Trigeminal and dorsal column nuclei projections to the anterior pretectal nucleus in the rat

The projections of the trigeminal (V) sensory nuclei (VSN) and the dorsal column nuclei (DCN) to the anterior pretectal nucleus (APT) of the rat were investigated by the use of anterograde and retrograde transport of wheat-germ agglutinin-conjugated horseradish peroxidase (WGA-HRP). Injections of WGA-HRP into the APT retrogradely labeled neurons in the contralateral VSN and DCN. The labeled neurons in the VSN were most concentrated in the rostral V subnucleus interpolaris (Vi), but were also found in caudal V subnucleus oralis (Vo). No labeled neurons were seen in V subnucleus caudalis. In the DCN, retrogradely labeled neurons were observed in rostral portions of both the cuneate (Cu) and gracile (Gr) nuclei. Injections of WGA-HRP into the rostral Vi or caudal Vo resulted in dense anterograde terminal labeling in the ventral two-thirds of the APT; the labeling was maximal in the ventromedial part of the caudal half of the APT and did not extend into its most rostral portion. Labeling resulting from injections of tracer into Cu or Gr was located primarily in the ventral half of the APT, was maximal in the mid-levels of the nucleus and extended into its rostral portions. These results indicate the existence of prominent somatosensory projections to the APT and are consistent with recent findings suggesting a role for the APT in sensorimotor integration.

Time course of serotonergic afferent plasticity within rat spinal trigeminal nucleus following infraorbital nerve transection

High-performance liquid chromatography with electrochemical detection (HPLC-ED) and immunocytochemistry were used to examine the time course of serotonergic afferent plasticity within trigeminal subnucleus interpolaris (SpVi) following infraorbital nerve (ION) transection in adult rats. Biochemical analysis was also performed in trigeminal subnucleus caudalis (SpVc) to examine the possibility of transient lesion-induced changes in this region. No significant changes in serotonin (5-HT) or 5-hydroxyindoleacetic acid (5-HIAA) concentration, or in density of 5-HT-immunoreactive (5-HTIR) axonal varicosities were observed in either subnucleus on the lesioned side, up to 51 days following ION cut. However, at 76–79 days post-lesion, a significant increase in 5-HT concentration was again demonstrated within SpVi.

Differential effects of peripheral damage on vibrissa-related patterns in trigeminal nucleus principalis, subnucleus interpolaris, and subnucleus caudalis

Histochemistry for cytochrome oxidase reveals a vibrissa-related pattern in trigeminal nucleus principalis, subnucleus interpolaris, and the magnocellular portion of subnucleus caudalis. This pattern is apparent in late fetal animals and is disrupted by transection of the infraorbital nerve on the day of birth. We recently reported results suggesting that the cytochrome oxidase pattern reflects primary afferent-induced clustering of second order neurons in all of these nuclei. If this conclusion is correct, it should follow that primary afferent lesions made after the cytochrome oxidase pattern became established in the brainstem might have little effect upon it. Accordingly, we transected the infraorbital nerve (the trigeminal branch that supplies the vibrissae) on postnatal days 0–10 and evaluated the vibrissa-related pattern in the brainstem with cytochrome oxidase histochemistry at varying intervals after these lesions. If the infraorbital nerve was sectioned on postnatal days 0–2, the vibrissa-related pattern was absent in trigeminal nucleus principalis, and both subnucleus interpolaris and caudalis. If such lesions were made after postnatal day 9, there was no appreciable effect upon the cytochrome oxidase pattern in any portion of the trigeminal brainstem complex. However, if lesions were made between postnatal days 3 and 8, the density and clarity of the cytochrome oxidase staining pattern were reduced in interpolaris and caudalis, but not in principalis. This difference was not due to differential transganglionic degeneration in these nuclei. Tracing with horseradish peroxidase demonstrated qualitatively equivalent primary afferent losses in principalis, interpolaris, and caudalis. Immunocytochemistry with a monoclonal antibody directed against parvalbumin also demonstrated a vibrissa-related pattern of cell bodies in principalis and interpolaris in rats killed on postnatal day 9 or later ages. The combination of retrograde tracing and immunocytochemistry revealed that the parvalbuminimmunoreactive neurons in principalis projected to thalamus while those in interpolaris were not labelled by tracer injections into the thalamus, midbrain, cerebellum or spinal cord. Infraorbital nerve transections made as late as postnatal day 8 resulted in a sharp decrease in the staining of parvalbumin-positive neurons in interpolaris, but not in principalis. Lesions made on postnatal day 10 had no qualitative effect upon parvalbumin-positive neurons in any portion of the trigeminal brainstem complex. The results of this study support the conclusion that the vibrissa-related cytochrome oxidase pattern in principalis becomes independent of primary afferent input at a very short interval after its initial appearance. In contrast, the patterns in more caudal portions of the trigeminal brainstem complex require maintenance of primary afferent input for a much longer postnatal period. The present results also provide further support for the conclusion that postsynaptic elements in the trigeminal brainstem complex may underlie the primary afferent-induced cytochrome oxidase pattern.

Excitatory amino acid binding sites in the trigeminal principal sensory and spinal trigeminal nuclei of the rat

Quantitative autoradiography was used to examine the density and distribution of excitatory amino acid (EAA) binding site subtypes in the principal sensory and spinal trigeminal nuclei of the rat trigeminal complex. The highest densities of N-methyl-d-aspartate (NMDA), α-amino-3-hydroxy-5-methylisoxazole-4-proprionic acid (AMPA), kainate and metabotropic receptors were found in the superficial laminae (I and II) of subnucleus caudalis, a region known to be densely innervated by primary afferent nociceptive terminals. Lower densities of EAA binding sites were observed in spinal subnuclei interpolaris and oralis and within the principal sensory nucleus. These results are consistent with the hypothesis that EAAs are involved in primary afferent nociceptive neurotransmission.

Parvalbumin and calbindin immunocytochemistry reveal functionally distinct cell groups and vibrissa-related patterns in the trigeminal brainstem complex of the adult rat.

Immunocytochemistry for calbindin (CA) and parvalbumin (PA) was combined with retrograde tracing from the thalamus, superior colliculus (SC), and cerebellum to define the ascending projections of neurons in the rat's trigeminal (V) brainstem complex that express immunoreactivity for these calcium binding proteins. Many PA-immunoreactive neurons were observed in trigeminal nucleus principalis (PrV). Many of these cells projected to thalamus and a few sent axons to SC. In ventral PrV, PA-immunoreactive neurons were arranged in a vibrissa-related pattern. A very small number of large CA-immunoreactive neurons were observed in dorsomedial PrV. None of these cells were labeled by our tracer deposits. Small neurons in V subnucleus oralis (SpO) were also immunoreactive for PA, but none were retrogradely labeled. A small percentage of the large neurons in SpO were CA-immunoreactive; many of these were retrogradely labeled by tracer injections in the thalamus and/or SC. In V subnucleus interpolaris (SpI), many small to medium sized cells were PA-positive and they were arrayed in a vibrissae-like pattern. None of these neurons were retrogradely labeled from any of the above-listed targets, but many were retrogradely labeled by tracer injections into ipsilateral PrV. SpI also contained many large CA-immunoreactive cells. Many of these projected to the thalamus and/or SC and some were also retrogradely labeled by tracer injections into ipsilateral PrV. In V subnucleus caudalis (SpC), very dark PA-immunoreactive neurons were located in the inner part of lamina II and less often in laminae I. Lightly labeled cells were located in the magnocellular laminae and formed vibrissa-related aggregates. None of these neurons were retrogradely labeled by our tracer injections. CA-immunoreactive cells were located throughout the depth of lamina II in SpC and smaller numbers were also visible in lamina I and layers III-V. A small percentage of the CA-positive cells in lamina I and in the magnocellular layers were retrogradely labeled from the thalamus. These data indicate that PA and CA antisera identify two cell populations in whisker-related regions of the V brainstem complex and that PA cells are somatotopically patterned in PrV, SpI, and SpC. These markers also distinguish two cell groups in superficial laminae of the medullary dorsal horn.

Response characteristics of tooth pulp-driven postsynaptic neurons in the spinal trigeminal subnucleus oralis of the cat.

Neuronal activity in the spinal trigeminal subnucleus oralis in response to electrical tooth stimulation was recorded in the anaesthetized cat in order to compare the electrophysiological characteristics of the oralis neurons with those of subnucleus caudalis and interpolaris neurons recorded in previous studies. The most sensitive oralis neurons had lower thresholds and shorter latencies than the most sensitive caudalis and interpolaris neurons. The thresholds of the oralis neurons were lower and their strength-duration curves flatter than those depicting liminal dental pain in man but similar to those depicting liminal jaw reflexes in the cat. Noxious conditioning stimulus elevated the threshold of only 1 of 10 neurons tested. The converging input from the skin and oral mucosa was from low-threshold mechanoreceptors. The results indicate that the response properties of the subnucleus oralis neurons differ significantly from those of other spinal subnuclei. Human pain thresholds cannot be explained by the liminal response properties of oralis neurons. These neurons might be important in the mediation of liminal reflex events evoked by dental stimuli.

Evidence for prenatal competition among the central arbors of trigeminal primary afferent neurons

Previous studies have shown that damage to vibrissa follicles in newborn rats and mice does not alter the brainstem representations of the remaining vibrissa as demonstrated by staining for mitochondrial enzymes such as cytochrome oxidase (CO) succinic dehydrogenase. This study asked whether this lack of effect might be due to the fact that the trigeminal primary afferents in rodents are already quite well developed at birth. We assessed this possibility by using CO staining the evaluate patterns in the brainstems of pre- and postnatal rats. A vibrissa-related pattern began to emerge in trigeminal nucleus principalis and subnucleus interpolaris (Spl) by embryonic day (E-) 19 and appeared fully developed by the day of birth (P-0). We also made partial lesions of the vibrissa pad on E-15-20 and on P-0, killed pups on P-5-7, and measured the size of the CO-stained patches in Spl on both sides of the brainstem. The correspondence between CO patches and clusters of primary afferent terminal arbors was verified in some animals by combining transganglionic horseradish peroxidase tracing and CO staining. Vibrissa pad damage on E-15-18 resulted in significant (20.1-36.9%) increases in the average area of the remaining CO patches in Spl ipsilateral to the lesion. Vibrissa pad damage on E-19, E-20, and P-0 produced small (6.2-8.9%), but insignificant, increases in patch size in Spl ipsilateral to the lesion. We used anatomical and electrophysiological methods to determine whether our lesions altered the trigeminal innervation of surviving vibrissa follicles. We recorded single trigeminal ganglion cells from 12 rats that sustained vibrissa pad lesion on E-17. As in normal rats, all of the 49 vibrissa-sensitive ganglion cells isolated in the lesioned animals were responsive to deflection of one and only one vibrissa. We also dissected 11 deep vibrissal nerves from intact follicles in adult rats that sustained fetal vibrissa pad damage on E-17, and counted numbers of myelinated axons in 1 microns plastic sections. These data were compared with counts from corresponding follicles on the intact side of the face. The average number of myelinated axons innervating follicles in the damaged vibrissa pads was 196.8 +/- 27.9, and that for the corresponding contralateral nerves was 194.6 +/- 25.7. These data suggest that competitive interactions among the central arbors of trigeminal primary afferents in fetal life may influence the development of central vibrissa representations and, further, that lesion-induced central changes need not be correlated with alterations in the peripheral innervation of undamaged follicles.