Central projections and somatotopic organisation of trigeminal primary afferents in pigeon (Columba livia)

Injections of cholera toxin B-chain conjugated to horseradish peroxidase into individual peripheral branches of the trigeminal nerve or into the trigeminal ganglion showed that an ascending trigeminal tract (TTA) terminated in distinct ventral and dorsal divisions of the principal sensory nucleus (PrVv and PrVd, respectively), and a descending tract (TTD) terminated within pars oralis, pars interpolaris, and pars caudalis divisions of the nucleus of TTD (nTTD) and within the dorsal horn of the first six cervical spinal segments. In PrVD, mandibular, ophthalmic, and maxillary projections were predominantly located dorsally, ventrally, and medially, respectively. In nTTD, mandibular projections lay dorsomedially, ophthalmic projections lay ventrolaterally, and maxillary projections lay in between. At caudal medullary and spinal levels, mandibular projections were situated medially, ophthalmic projections were situated laterally, and maxillary projections were situated centrally. The terminations within the dorsal horn were most dense in laminae III and IV and were least dense in lamina II, with laminae III-IV also receiving topographically organised contralateral projections. Extratrigeminal projections were mainly to the external cuneate nucleus by way of a lateral descending trigeminal tract (ITTD; Dubbeldam and Karten [1978] J. Comp. Neurol. 180:661–678) and to the region of the tract of Lissauer and lamina I of the dorsal horn. Other projections were to a region medial to the apex of pars interpolaris, to the nuclei ventrolateralis anterior (Vla) and presulcalis anterior (Pas) of the solitary complex, and sparsely to the lateral reticular formation (plexus of Horsley) ventral to TTD. No projections were seen to the trigeminal motor nuclei or to the cerebellum. © 1996 Wiley-Liss, Inc.     

The central projections of trigeminal primary afferent neurons in the cat as determined by the tranganglionic transport of horseradish peroxidase

The central projections of five peripheral branches of the trigeminal nerve were investigated by the method of transganglionic transport of horseradish peroxidase (HRP). In separate animals, the corneal, supraorbital, infraorbital, mental, or inferior alveolar branches were transected and soaked in concentrated solutions of HRP. Forty-eight to 72 hours after surgery, the brainstem, upper cervical spinal cord, and trigeminal ganglia were perfusion-fixed and processed according to the tetramethylbenzidine technique. The results show that trigeminal primary afferent neurons which innervate the cornea project mainly to the levels of caudal pars interpolaris and caudal pars caudalis. In contrast, trigeminal primary afferent neurons whose peripheral processes course through the supraorbital, infraorbital, or mental nerves project most heavily to the trigeminal main sensory nucleus, pars interpolaris, and the rostrocaudal middle three-fifths of pars caudalis. Trigeminal primary afferent neurons which give origin to the inferior alveolar nerve project heavily and in approximately equal numbers of all rostrocaudal levels of the trigeminal brainstem nuclear complex (TBNC). A small number of fibers from each of the latter four cell populations project directly to the contralateral C1-C2 dorsal horn. A small number of fibers from each cell population studied end in the reticular formation immediately adjacent to the spinal nucleus of V. It is concluded that the cornea and facial skin regions of the cat are represented nonuniformly along the rostrocaudal length of the TBNC.     

Efferent projections from temperature sensitive recording loci within the marginal zone of the nucleus caudalis of the spinal trigeminal complex in the cat

The efferent projections from nucleus caudalis of the spinal trigeminal complex in cats were studied with retrograde and anterograde axonal transport techniques combined with localization of recording sites in the thalamus and marginal zone of nucleus caudalis to innocuous skin cooling. Results showed brainstem projections from nucleus caudalis to rostral levels of the spinal trigeminal complex, to the ventral division of the principal trigeminal nucleus, the parabrachial nucleus, cranial motor nuclei 7 and 12, solitary complex, contralateral dorsal inferior olivary nucleus, portions of the lateral reticular formation, upper cervical spinal dorsal horn and, lateral cervical nucleus. Projections to the thalamus included: a dorsomedial region of VPM (bilaterally) and to the main part of VPM and PO contralaterally. Neuronal activity was recorded in the dorsomedial region of VPM to cooling the ipsilateral tongue. HRP injections in this thalamic region retrogradely labeled marginal neurons in nucleus caudalis. These results show that marginal neurons of nucleus caudalis provide a trigeminal equivalent of spinothalamic projections to the ventroposterior nucleus in cats.     

Corneal sensory pathway in the rat: a horseradish peroxidase tracing study

The methods of transganglionic transport of horseradish peroxidase (HRP) and horseradish peroxidase--wheat germ agglutinin (HRP-WGA) were used to determine the location within the trigeminal ganglion of the primary afferent neurons that innervate the rat central cornea, and the brainstem and spinal cord termination sites of these cells. In each of 18 animals, solutions of HRP or HRP-WGA were applied to the scarified corneal surface and allowed to infiltrate into the corneal epithelium and stroma for 15 minutes. Postmortem examination of the corneal whole mounts from the experimental animals, and of corneas and neural tissues from several control animals, showed that the HRP/HRP-WGA remained confined to the central cornea with no spread into adjacent intra- or extraorbital tissues. HRP-labeled corneal afferent somata were located in the dorsal part of the ophthalmic region of the ipsilateral trigeminal ganglion. The central fibers of the corneal afferent neurons projected very heavily to interstitial nuclei of Cajal in the spinal tract of V at the level of caudal pars interpolaris and rostral pars caudalis, lightly to the pars caudalis/C1 transition zone, and sparsely to the dorsal horn of spinal cord segments C1-C3. The trigeminal main sensory nucleus, pars oralis, the rostral three-fourths of pars interpolaris, and an extensive midregion of pars caudalis were totally devoid of reaction product. Terminal fields in caudal pars caudalis and in the spinal cord dorsal horn were concentrated largely in the outer half of lamina II, with lesser accumulations in lamina I, the deeper half of lamina II, and in lamina III. The present study demonstrates for the first time by means of an anatomical tracing procedure the brainstem termination sites of corneal afferent neurons in the rat. The patchy, discontinuous nature of the corneal afferent projection to the caudal trigeminal brainstem nuclear complex (TBNC), and the total lack of corneal projections to rostral subdivisions of the TBNC, provide an exception to the general rule of trigeminal organization in which most areas of the head and face are represented as continuous columns throughout the rostrocaudal extent of the ipsilateral TBNC.     

The central projections of tooth pulp afferent neurons in the rat as determined by the transganglionic transport of horseradish peroxidase

Transganglionic transport of horseradish peroxidase (HRP) or horseradish peroxidase-wheat germ agglutinin conjugate (HRP-WGA) was used to map in detail the central projections of trigeminal primary afferent neurons that innervate the dental pulp organ of the rat. In each of ten animals, 0.5-2.0 microliters of enzyme solution was injected into the pulp chamber of the first maxillary molar tooth. Postmortem examination of the decalcified teeth in all cases showed that the HRP/HRP-WGA remained confined to the pulp chamber and pulp roots, with no spread of enzyme into periapical tissues. HRP-labeled tooth pulp afferent fibers projected to all four rostrocaudal subdivisions of the ipsilateral trigeminal brainstem nuclear complex (TBNC) and to the upper cervical spinal cord. The labeled terminal fields formed a column that stretched relatively uninterrupted from just caudal to the rostromedial tip of the trigeminal principal sensory nucleus to at least the C2 segment of the spinal cord. The density of the afferent projection varied markedly from one rostrocaudal level of the TBNC to the next but was heaviest in an area encompassing the caudal one-half of the principal sensory nucleus and the rostral two-thirds of pars oralis. Fibers projected only lightly to pars caudalis, where they terminated preferentially in laminae I, IIa, and the junctional zone between laminae IV and V. HRP-labeled terminals in C1 and C2 were located almost exclusively in laminae I. In the dorsoventral axis, the terminal fields in the TBNC were located in a surprisingly dorsal part of the complex, well within what has been shown by others to be largely an area of termination for mandibular division fibers. Most fibers ended in medial parts of the TBNC, with the exception of two modestly labeled terminal fields located in the lateral aspects of rostral pars oralis and rostral pars caudalis. No labeled fibers terminated in the contralateral TBNC or contralateral cervical spinal cord.     

The ascending input to the midbrain periaqueductal gray of the primate

To obtain a comprehensive map of the brainstem and spinal cord areas that project to the mesencephalic central gray small injections of hors-radish peroxidase were made into various regions of the periaqueductal gray in a series of monkeys. Despite the fact that different regions of the central gray were injected in separate animals, the majority of the brainstem areas containing retrogradely filled neurons remained the same. Labeled neurons were observed in the superior colliculus, periaqueductal gray, lateral parabrachial, locus coeruleus, nucleus raphe magnus and pallidus, and a variety of brainstem reticular nuclei. In contrast to labeled brainstem areas, where labeled neurons were present predominantly ipsilateral to the injection site, the spinal trigeminal nucleus pars caudalis and the spinal cord displayed labeled cells chiefly on the side contralateral to the injection. Also in contrast to the labeled brainstem sites, where medial and lateral injection sites produced a similar pattern of labeling, medial injections in the PAG labeled almost exclusively neurons in the deep laminae (V-X) in the spinal trigeminal nucleus pars caudalis and spinal cord while more lateral injections labeled neurons in both the deep (V-X) and superficial (I) laminae. No consistent differences were noted in the location of labeled neurons in either brainstem or spinal sites after dorsal vs. ventral injections or caudal vs. rostral injection sites. The present study has demonstrated that the central gray receives afferent projections from a number of brainstem and spinal areas which are known to be involved in the modulation andor conduction of nociception, while other inputs are probably involved in the regulation of visceral functions. These data support the hypothesis that the mesencephalic periaqueductal gray functions as a visceral, nociceptive, and cognitive integrator.     

Primary afferent projections from the upper respiratory tract in the muskrat.

The central projections of the ethmoidal, glossopharyngeal, and superior laryngeal nerves were determined in the muskrat by use of the transganglionic transport of a mixture of horseradish peroxidase (HRP) and wheat germ agglutinin (WGA)-HRP. The ethmoidal nerve projected to discrete areas in all subdivisions of the ipsilateral trigeminal sensory complex. Reaction product was focused in ventromedial portions of the principal nucleus, subnucleus oralis, and subnucleus interpolaris. The subnucleus oralis also contained sparse reaction product in its dorsomedial part. Projections were dense to ventrolateral parts of laminae I and II of the rostral medullary dorsal horn, with sparser projections to lamina V. Label in laminae I and V extended into the cervical dorsal horn. A few labeled fibers were followed to the contralateral dorsal horn. The interstitial neuropil of the ventral paratrigeminal nucleus was densely labeled. Extratrigeminal primary afferent projections in ethmoidal nerve cases involved the K?lliker-Fuse nucleus and ventrolateral part of the parabrachial nucleus, the reticular formation surrounding the rostral ambiguous complex, and the dorsal reticular formation of the closed medulla. Retrograde labeling in the brain was observed in only the mesencephalic trigeminal nucleus in these cases. The cervical trunk of the glossopharyngeal and superior laryngeal nerves also projected to the trigeminal sensory complex, but almost exclusively to its caudal parts. These nerves terminated in the dorsal and ventral paratrigeminal nuclei as well as lamina I of the medullary and cervical dorsal horns. Lamina V received sparse projections. The glossopharyngeal and superior laryngeal nerves projected to the ipsilateral solitary complex at all levels extending from the caudal facial nucleus to the cervical spinal cord. At the level of the obex, these nerves projected densely to ipsilateral areas ventral and ventromedial to the solitary tract. Additional ipsilateral projections were observed along the dorsolateral border of the solitary complex. Near the obex and caudally, the commissural area was labeled bilaterally. Labeled fibers from the solitary tract projected into the caudal reticular formation bilaterally, especially when the cervical trunk of the glossopharyngeal nerve received tracer. Labeled fibers descending further in the solitary tract gradually shifted toward the base of the cervical dorsal horn. The labeled fibers left the solitary tract and entered the spinal trigeminal tract at these levels. Retrogradely labeled cells were observed in the ambiguous complex, especially rostrally, and in the rostral dorsal vagal nucleus after application of HRP and WGA-HRP to either the glossopharyngeal or superior laryngeal nerves. In glossopharyngeal nerve cases, retrogradely labeled neurons also were seen in the inferior salivatory nucleus.(ABSTRACT TRUNCATED AT 400 WORDS)     

Central projections and trigeminal ganglion location of corneal afferent neurons in the monkey, Macaca fascicularis

The method of transganglionic transport of horseradish peroxidase-wheat germ agglutinin conjugate (HRP-WGA) was used to determine the location within the monkey trigeminal ganglion of the primary afferent neurons that innervate the cornea, and the brainstem and spinal cord termination sites of these cells. In each of four animals. Gelfoam pledgets were saturated with 2% HRP-WGA in saline and applied to the scratched surface of the central cornea for 30 minutes. Postmortem examination of the corneal whole mounts revealed that the tracer solution remained confined to approximately the central one-fourth of the cornea with no spread into the peripheral cornea or limbus. Seventy-two to 96 hours after tracer application, 126-242 labeled cell bodies were observed in the medial region of the ipsilateral trigeminal ganglion. The majority of neurons were concentrated in an area of the ganglion that lay directly caudal to the entering fibers of the ophthalmic nerve, but smaller numbers of cells lay somewhat more laterally, near the region where the ophthalmic and maxillary nerves come together. A very small number of neurons in one animal innervated the cornea by sending their fibers into the maxillary nerve. HRP-WGA-labeled terminal fields were present to some extent in all four major rostrocaudal subdivisions of the ipsilateral trigeminal brainstem nuclear complex (TBNC), but the size of the terminal fields and the intensity of labeling differed markedly from one level of the TBNC to the next. Labeled fibers projected heavily to the transitional zone between caudal pars interpolaris and rostral pars caudalis (i.e., the "periobex" region of the TBNC) and moderately to the trigeminal main sensory nucleus, pars oralis, and caudal pars caudalis at the level of the pyramidal decussation. Remaining areas of the TBNC, including rostral pars interpolaris and the midlevel of pars caudalis, received few, if any, corneal afferent projections. Occasional labeled fibers were observed in the dorsal horn of C1 and in the rostral half of C2. It is hoped that data generated in the current investigation of nonhuman primates will contribute to a better understanding of the neural substrates that subserve corneal sensation and the blink reflex in humans.     

Brain stem terminations of the trigeminal and upper spinal ganglia innervation of the cerebrovascular system: WGA-HRP transganglionic study

The central projections of the nerve fibers innervating the middle cerebral and basilar arteries were investigated by transganglionic tracing of wheat germ agglutinin conjugated with horseradish peroxidase (WGA-HRP) in the rat. WGA-HRP was applied to the exposed basilar and/or middle cerebral arteries. Sections of the brain, trigeminal and upper spinal ganglia were reacted with tetramethylbenzidine for detection of the tracer. The results demonstrate that trigeminal neurons that innervate the middle cerebral artery project to the trigeminal main sensory nucleus, pars oralis, and the dorsocaudal two-fifths of pars interpolaris of the trigeminal brain stem nuclear complex. Terminals were also visible in the ipsilateral nucleus motorius dorsalis nervi vagi (dmnX) and in the lateral nucleus tractus solitarius (nTs) bilaterally at the level of the obex. The ventral periaqueductal gray, including the dorsal raphe and C2 dorsal horn, were also innervated by nerve fibers from the middle cerebral artery. Ipsilateral trigeminal rhizotomy prior to WGA-HRP application over the middle cerebral artery impeded the visualization of nerve terminations throughout the brain stem. Pretreatment with capsaicin reduced the density of labeled neurons and terminals within the trigeminal ganglion and the brain stem, respectively, following WGA-HRP application over the middle cerebral artery. Basilar artery fibers terminate in the C2 dorsal horn, the cuneate nuclei, dmnX, and nTs bilaterally. A few projections were also labeled in the ventral periaqueductal gray. Unilateral upper two spinal dorsal rhizotomy prior to WGA-HRP application over the exposed basilar artery resulted in terminal labeling within the C2 dorsal horn, the cuneate nucleus, dmnX, and nTs contralateral to the rhizotomy, whereas the ipsilateral side was devoid of any labeling. Bilateral superior cervical ganglionectomy prior to WGA-HRP administration to the middle cerebral and basilar arteries did not alter the visualization of nerve terminations throughout the brain stem.     

Localization of met-enkephalin-like immunoreactivity within pain-related nuclei of cervical spinal cord, brainstem and midbrain in the cat

Met-enkephalin immunoreactivity was investigated with an indirect immunoperoxidase technique in the cervical spinal cord, brainstem and midbrain of the cat, paying special attention to pain-related nuclei. Different technical conditions were used to reveal preferentially met-enkephalin-containing fibres and terminals or perikarya. Immunoreactive fibres and terminals were revealed optimally in sections from control animals incubated with detergent (Triton X-100). Immunoreactive perikarya were revealed in colchicine treated animals. Comparison between different routes of administration showed that local injections of colchicine are needed to reveal optimally immunoreactive perikarya in nuclei located far from the ventricles. Met-enkephalin-containing fibres and terminals are widely distributed in the posterior brain and spinal cord. The densest network of immunoreactive fibres are observed in the superficial layers of the cervical spinal cord and the caudal trigeminal nucleus, in the nucleus of the solitary tract, the nucleus of the facial nerve, the nucleus of the prepositus hypoglossi, the nucleus raphe pallidus, the medial vestibular nucleus, the interpedoncular nucleus and the substantia nigra. A moderate staining of fibres is observed in various nuclei including the ventral horn of the spinal cord and caudal trigeminal nucleus, the brainstem and midbrain reticular formation, the inferior olivary complex, the nucleus of the descending trigeminal tract and the periaqueductal grey. Met-enkephalin-containing perikarya are present in all the nuclei cited before, except in the inferior olivary complex. The densest aggregation of enkephalin-like perikarya is observed in the nucleus raphe magnus, nucleus raphe obscurus, nucleus raphe pallidus, nucleus reticularis gigantocellularis pars α and nucleus reticularis lateralis. The general distribution of enkephalin-containing structures in the cervical spinal cord, brainstem and midbrain of the cat appears very similar to that of the rat except in the substantia nigra where met-enkephalin cell bodies are found in the cat but not in the rat. In particular the pain-related nuclei present a similar distribution of the peptide in the two species; however, met-enkephalin-containing cell bodies are much more numerous in the cat than in the rat (notably in the reticular formation). Similar types of metenkephalin innervation occur in the dorsal and intermediate grey of the spinal cord and of the caudal trigeminal nucleus supporting further that the functional organizations of these regions are closely related.     

Spinal and trigeminal projections to the nucleus of the solitary tract: a possible substrate for somatovisceral and viscerovisceral reflex activation

This study used the retrograde transport of a protein-gold complex to examine the distribution of spinal cord and trigeminal nucleus caudalis neurons that project to the nucleus of the solitary tract (NST) in the rat. In the spinal grey matter, retrogradely labeled cells were common in the marginal zone (lamina I), in the lateral spinal nucleus of the dorsolateral funiculus, in the reticular part of the neck of the dorsal horn (lamina V), around the central canal (lamina X), and in the region of the thoracic and sacral autonomic cell columns. The pattern of labeling closely resembled that seen for the cells at the origin of the spinomesencephalic tract and shared some features with that of the spinoreticular and spinothalamic tracts. Labeled cells in lamina IV of the dorsal horn were only observed when injections spread dorsally, into the dorsal column nuclei, and are thus not considered to be at the origin of the spinosolitary tract. They are probably neurons of the postsynaptic fibers of the dorsal column. Retrogradely labeled cells were also numerous in the superficial laminae of the trigeminal nucleus caudalis, through its rostrocaudal extent. The pattern of marginal cell labeling appeared to be continuous with that of labeled neurons in the paratrigeminal nucleus, located in the descending tract of trigeminal nerve. Since the NST is an important relay for visceral afferents from both the glossopharyngeal and vagus nerves, we suggest that the spinal and trigeminal neurons that project to the NST may be part of a larger system that integrates somatic and visceral afferent inputs from wide areas of the body. The projections may underlie somatovisceral and/or viscerovisceral reflexes, perhaps with a significant afferent nociceptive component.     

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

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

Central distribution of trigeminal and upper cervical primary afferents in the rat studied by anterograde transport of horseradish peroxidase conjugated to wheat germ agglutinin

Injections of WGA-HRP were made in the rat trigeminal ganglion and C1-3 dorsal root ganglia (DRGs) to study the central projection patterns and their relations to each other. Trigeminal ganglion injections resulted in heavy terminal labeling in all trigeminal sensory nuclei. Prominent labeling was also observed in the solitary tract nucleus and in the medial parts of the dorsal horn at C1-3 levels, but labeling could be followed caudally to the C7 segment. Contralateral trigeminal projections were found in the nucleus caudalis and in the dorsal horn at C1-3 levels. The C1 DRG was found to be inconstant in the rat. When it was present, small amounts of terminal labeling were found in the external cuneate nucleus (ECN) and the central cervical nucleus (CCN). No dorsal horn projections were seen from the C1 DRG. Injections in the C2 DRG resulted in heavy labeling in the ECN, nucleus X, CCN, and dorsal horn, where it was mainly located in lateral areas. Labeling could be followed caudally to the Th 7 segment. C2 DRG projections also appeared in the cuneate nucleus (Cun), in all the trigeminal sensory nuclei, and in the spinal, medial, and lateral vestibular nuclei. A small C2 DRG projection was observed in the ventral cochlear nucleus. C3 DRG injections resulted in heavy labeling in both medial middle and lateral parts of the dorsal horn, in the ECN, and in nucleus X, whereas the labeling in the CCN was somewhat weaker. Smaller projections were seen to trigeminal nuclei, Cun, and the column of Clarke. Comparisons of the central projection fields of trigeminal and upper cervical primary afferents indicated a somatotopic organization but with a certain degree of overlap.     

Trigemino-reticulo-facial and trigemino-reticulo-hypoglossal pathways in the rat

This study was undertaken to identify premotor neurons in the pontomedullary reticular formation serving as relay neurons between the sensory trigeminal complex and the motor nuclei of the VIIth and XIIth nerves. Trigeminoreticular projections were first investigated after injections of anterogradely transported tracers (biotinylated dextran amine, biocytin) into single subdivisions of the sensory trigeminal complex. The results show that the trigeminoreticular projections were abundant from the pars interpolaris (5i) and caudalis (5c) and moderate from pars oralis (5o) of the spinal trigeminal nucleus. Injections into the 5i and 5c produce dense anterograde labeling (1) in the dorsal medullary reticular field; (2) in the parvocellular reticular field, medially adjacent to the 5i; and (3) more rostral in the region dorsal and lateral to the superior olivary nucleus. Some labeled terminals were also found in the intermediate reticular field, whereas only light anterograde labeling was observed in the gigantocellular and oral pontine reticular formation. The 5o sends fibers and terminals throughout the whole reticular formation, with no clear preferential projections within a particular field. Only light projections originated from the principal nucleus (5P). In a second series of experiments, we examined whether premotor neurons in the reticular formation are afferented by trigeminal fibers. Double labeling was performed by injection of an anterograde tracer in the 5i and 5c and retrograde tracer (gold-horseradish peroxidase complex) into the VII or the XII motor nucleus on the same side. Retrogradely labeled neurons in contact with anterogradely labeled boutons were found throughout the reticular formation with predominance in the parvocellular and intermediate reticular fields. These experiments demonstrate the existence of trigeminal disynaptic influences, via reticular neurons of the pontomedullary reticular formation, in the control of orofacial motor behaviors.     

NADPH-diaphorase in the developing rat: Lower brainstem and cervical spinal cord,with special reference to the trigemino-solitary complex

A previous study indicated that in adult rat, a distinctive neuronal group in the dorsomedial division of the subnucleus oralis of the spinal trigeminal nucleus (SpVo) and the rostrolateral part of the nucleus of the solitary tract (Sn) is stained for nicotinamide adenine dinucleotide phosphate-diaphorase (NADPH-d), and suggested that the labeled structures are involved with sensorimotor reflexive functions. This study aimed to characterize the developmental expression of NADPH-d in SpVo and Sn, including other areas of the lower brainstem and cervical spinal cord, by means of the enzyme histochemical staining technique, from the prenatal through the postnatal period. On embryonic day 12 (E12), no neurons in the brain were stained for NADPH-d, whereas blood vessels were stained. Labeling in the vessels was consistently present throughout pre- and postnatal periods but decreased with development. On E15, labeled neurons appeared in the dorsomedial part of SpVo and the rostrolateral part of Sn, but not in the other nuclei. The labeled neurons in both nuclei increased in numbers drastically to E17. Postnatally, they tended to increase gradually in Sn, but to decrease slightly in SpVo. The cell size of labeled neurons reached a plateau at E17 in SpVo, but at postnatal day 4 (P4) in Sn. In other nuclei on E17, labeling appeared in the lateral paragigantocellular reticular, intermediate reticular, medullary reticular, pedunculopontine tegmental, and spinal vestibular nuclei, and laminae V, VI, and X of the cervical spinal cord. On E20 and P0, labeling appeared in the dorsal column, laterodorsal tegmental, raphe obscurus, parvocellular reticular, ventral gigantocellular reticular, and parahypoglossal nuclei, and laminae IX of the cervical spinal cord. On P4, labeling appeared in the parabrachial and median raphe nuclei, medial and caudolateral Sn, the magnocellular zone of subnucleus caudalis of the spinal trigeminal nucleus (SpVc), and laminae III/IV of the cervical spinal cord. On P10, labeling appeared in the paratrigeminal and dorsal raphe nuclei, the superficial zone of SpVc, and laminae I/II of the cervical spinal cord. No newly labeled neurons appeared in any nuclei after P14. The very early appearance of NADPH-d staining in SpVo and Sn, which precedes the appearance of NADPH-d elsewhere in the brainstem, suggests that the nitric oxide (NO)/cyclic guanosine monophosphate (cGMP) system has an important role for primitive orofacial sensorimotor reflexive functions. Furthermore, the pattern of developmental expression of NADPH-d in SpVo and Sn suggests that the NO/cGMP system is organized in a distinct manner in different nuclei. © 1996 Wiley-Liss, Inc.     

Localization of trigeminal, spinal, and reticular neurons involved in the rat blink reflex

Electrical stimulation of the supraorbital nerve (SO) induces eyelid closure by activation of orbicularis oculi muscle motoneurons located in the facial motor nucleus (VII). Neurons involved in brainstem central pathways implicated in rat blink reflex were localized by analyzing c-Fos protein expression after SO stimulation in conjunction with tracing experiments. A retrograde tracer (gold-horseradish peroxidase [HRP]) was injected into the VII. The distribution patterns of activated c-Fos-immunoreactive neurons and of neurons exhibiting both c-Fos immunoreactivity and gold-HRP labeling were determined in the sensory trigeminal complex (STC), the cervical spinal cord (C1), and the pontomedullary reticular formation. Within the STC, c-Fos immunoreactivity labeled neurons in the ipsilateral ventral part of the principal nucleus, the pars oralis and interpolaris, and bilaterally in the pars caudalis. Colocalization of gold-HRP and c-Fos immunoreactivity was observed in neurons of ventral pars caudalis layers I-IV and ventral pars interpolaris. In C1, SO stimulation revealed c-Fos neurons in laminae I-V. After additional injections in VII, the double-labeled c-Fos/gold-HRP neurons were concentrated in laminae IV and V. Although c-Fos neurons were found throughout the pontomedullary reticular formation, most appeared rostrally around the motor trigeminal nucleus and in the ventral parvocellular reticular nucleus medial to the fiber bundles of the seventh nerve. Caudally, c-Fos neurons were in the lateral portion of the dorsal medullary reticular field. In addition, these reticular areas contained double-labeled neurons in electrically stimulated rats that had received gold-HRP injections in the VII. The presence of double-labeled neurons in the STC, C1, and the reticular formation implies that these neurons receive sensory information from eyelids and project to the VII. These double-labeled neurons could then be involved in di- or trisynaptic pathways contributing to the blink reflex.     

Trigeminal-reticular connections: Possible pathways for nociception-induced cardiovascular reflex responses in the rat

Cardiovascular regulatory neurons of the ventral medulla and pons are thought to have an important role in the mediation of trigeminal nociception-induced reflex cardiovascular responses. However, the neural pathways that link the spinal trigeminal nucleus with ventral medullary and pontine autonomic cell groups are poorly understood. The present study utilized injections of the highly sensitive anterograde tracer substance biotinylated dextran combined with immunocytochemistry for tyrosine hydroxylase, the synthesizing enzyme for catecholamines, to investigate the distribution and morphology of projections from the spinal trigeminal subnucleus caudalis to ventral medullary and pontine catecholaminergic cell groups. Injection of biotylinated dextran into the dorsal subnucleus caudalis produced dense anterograde labeling in dorsal regions of the medullary and pontine reticular formation including the dorsal medullary reticular field, the parvicellular reticular field, and the parvicellular reticular field pars anterior. In the ventral medullary and pontine reticular formation, light anterograde labeling tended to be distributed in close proximity to the distal dendrites of catecholaminergic neurons located in the C1, A1, and A5 regions. Injections of anterograde tracer into the dorsal medullary reticular field produced dense anterograde labeling in the ventral medullary and pontine reticular formation. Numerous terminal-like varicosities were observed in close proximity to catecholaminergic neurons located in the C1, A1, and A5 regions. These data suggest that trigeminal pain-induced reflex cardiovascular responses involve indirect projections that terminate in the dorsal medullary and pontine reticular formation before reaching ventral medullary and pontine catecholaminergic cell groups known to be involved in cardiovascular regulation. J. Comp. Neurol. 391:526–544, 1998. © 1998 Wiley-Liss, Inc.     

Efferent connections of the subthalamic region in the rat. II. The zona incerta

The efferent connections of the zona incerta (ZI) were studied experimentally in the rat by the aid of the autoradiographic tracer technique.Small microelectrophoretic injections of tritiated proline and leucine practically confined to the ZI were found to label a widespread, predominantly ipsilateral system of descending and ascending fibers distributed to reticular structures of the brain stem (mesencephalic reticular formation, nucleus tegmenti pedunculopontinus pars compacta, parabrachial area, nuclei reticularis pontis oralis, pontis caudalis, gigantocellularis and medullae oblongatae, pars ventralis), precerebellar nuclei (nucleus reticularis tegmenti pontis, pontine nuclei and inferior olivary complex), the middle and deep layers of the superior colliculus, the pretectum (anterior, posterior and medial pretectal nuclei), perioculomotor nuclei (interstitial nucleus of Cajal, nucleus of Darkschewitsch and nuclei of the posterior commissure), the parvocellular portion of the red nucleus, the central gray substance, the nucleus tegmenti dorsalis lateralis, the ventral horn of the cervical spinal cord, non-specific thalamic nuclei (parafascicular, centralis medius, paracentralis centralis lateralis and ventromedial thalamic nuclei, nucleus reuniens), basal ganglia (entopeduncular nucleus and globus pallidus), hypothalamic structures (posterior hypothalamic nucleus, dorsal and lateral hypothalamic areas), and a subpallidal district of the substantia innominata. Isotope injections centered in Forel's field H₁ resulted in the labeling of a similar set of projections. Some of the possible functional correlates of these connections are briefly discussed.     

Contralateral projections of trigeminal mandibular primary afferents in the guinea pig as seen by transganglionic transport of horseradish peroxidase

Transganglionic transport of horseradish peroxidase (HRP) was used to investigate contralateral projections of trigeminal mandibular fibers in the guinea pig. After application of HRP to the buccal, lingual, auriculotemporal, mylohyoid, mental and inferior alveolar nerves, crossing fibers and contralateral endings were found in the caudal region of the nucleus of the solitary tract (most of these belonging to the buccal and lingual nerves), the dorsomedial region of the subnucleus caudalis of the trigeminal sensory nuclear complex (TSNC), and the dorsal horns of the first 5 cervical spinal cord segments (C1-C5). The greatest numbers of crossing fibers in the medullary and cervical dorsal horn segments belonged to the mental and mylohyoid nerves, though these nerves did not project contralaterally to C4-C5. Contralateral buccal and lingual endings were scattered sparsely from the subnucleus caudalis to C5, and only very few contralateral auriculotemporal terminals were observed. Though laminae I-V of the dorsomedial region of the medullary and cervical dorsal horns all exhibited contralateral endings of the mental and mylohyoid nerves, most such endings were found in laminae IIi-III, followed by lamina IV, which suggests their involvement in the reception of mechanical stimuli and in the sensory motor reflexes of the orofacial region. The contralateral buccal and lingual terminals were distributed somatotopically in the first 5 cervical cord segments, with the lingual endings rostral to the buccal terminals within each segment. In C4 and C5 lingual endings appeared exclusively in laminae I and IIo, suggesting that like the ipsilateral lingual projections at this level, which also terminate in these laminae, they may be involved in pain and temperature sensation.     

Somatosensory projection to the mesencephalon: an anatomical study in the monkey

The terminal areas and cells of origin of the somatosensory projection to the mesencephalon in the monkey were investigated by the intraaxonal transport method. Following injection of wheat germ agglutinin-horseradish peroxidase conjugate (WGA-HRP) into the spinal enlargements, the lateral cervical nucleus (LCN), the dorsal column nuclei (DCN), or the spinal trigeminal nucleus, anterograde labeling was observed in several regions of the mid-brain. (1) Injection of tracer into the spinal enlargements resulted in dense terminal labeling in the parabrachial nucleus (PBN) and the periaqueductal gray matter (PAG); moderate termination was observed in the intercollicular nucleus (Inc), the intermediate and deep gray layers of the superior colliculus (SGI, SGP), the posterior pretectal nucleus (PTP), and the nucleus of Darkschewitsch (D); and scattered terminal fibers were seen in the cuneiform nucleus (CNF) and the pars compacta of the anterior pretectal nucleus (PTAc). The projections from the cervical enlargement to PAG, Inc, and the superior colliculus terminated more rostrally than those from the lumbar segments, indicating a somatotopic organization. (2) Terminal labeling after injection of tracer into LCN was found mainly in Inc, SGI, and SGP, but sparse labeling was also observed in the nucleus of the brachium of the inferior colliculus (BIN), PAG, PBN, PTP, and D. (3) The projection from DCN terminated densely in the external and pericentral nuclei of the inferior colliculus (ICX, ICP), Inc, SGI, SGP, PTP, PTAc, the nucleus ruber, and D, and weak terminal labeling was seen in BIN, PAG, and PBN. Comparisons of the anterograde labeling following injections involving both the gracile nucleus and the cuneate nucleus with that after injection restricted to the gracile nucleus alone suggested a somatotopic termination pattern in Inc, the superior colliculus, and the pretectal nuclei. (4) The patterns of projection from the laminar and alaminar parts of the spinal trigeminal nucleus differed: injection of tracer into the caudal part of the alaminar spinal trigeminal nucleus (nucleus interpolaris) resulted in dense anterograde labeling in SGI and SGP, moderate termination in Inc, and minor projections to PBN, PAG, and PTP, whereas after tracer injection into the laminar trigeminal nucleus (nucleus caudalis) terminal labeling was present only in PBN and PAG. Following injection of tracer into the midbrain terminal areas retrogradely labeled neurons were found in the spinal cord, LCN, DCN, and the spinal trigeminal nucleus, with the majority of labeled cells situated on the side contralateral to the injection site.(ABSTRACT TRUNCATED AT 400 WORDS)