Brainstem projections from recipient zones of the anterior ethmoidal nerve in the medullary dorsal horn.

Stimulation of the anterior ethmoidal nerve or the nasal mucosa induces cardiorespiratory responses similar to those seen in diving mammals. We have utilized the transganglionic transport of a cocktail of horseradish peroxidase conjugates and anterograde and retrograde tract tracing techniques to elucidate pathways which may be important for these responses in the rat. Label was seen throughout the trigeminal sensory complex after the horseradish peroxidase conjugates were applied to the anterior ethmoidal nerve peripherally. Reaction product was most dense in the medullary dorsal horn, especially in laminae I and II. Injections were made of biotinylated dextran amine into the recipient zones of the medullary dorsal horn from the anterior ethmoidal nerve, and the anterogradely transported label documented. Label was found in many brainstem areas, but fibers with varicosities were noted in specific subdivisions of the nucleus tractus solitarii and parabrachial nucleus, as well as parts of the caudal and rostral ventrolateral medulla and A5 (noradrenergic cell group in ventrolateral pons) area. The retrograde transport of FluoroGold into the medullary dorsal horn after injections into these areas showed most neurons in laminae I, II, and V. Label was especially dense in areas which received primary afferent fibers from the anterior ethmoidal nerve. These data identify potential neural circuits for the diving response of the rat.

The central termination of sensory fibers from nerves to the gastrocnemius muscle of the rat.

Peripheral nerves innervating muscles have sensory fibers that relay information into the CNS information about proprioception, pain, and the metabolic state of the muscle. The present study shows the primary afferent projections into the spinal cord of the nerves innervating the gastrocnemius muscle of the rat using the transganglionic transport of a cocktail of horseradish peroxidase (HRP) conjugated to cholera toxin and wheat germ agglutinin; these markers have been shown to label large and small fibers, respectively. A dense projection into lamina I of the lumbar dorsal horn and a more moderate projection into lamina V were seen. Moreover, dense reaction product was found in the most medial aspect of lamina II, especially lamina II inner part, and less in lamina III and IV of levels L3-L5. Lamina VI had dense reaction product from the rostral sacral levels of the spinal cord that continued into Clarke's column at rostral lumbar levels. The nucleus gracilis also was labeled. Other nerves emerging from the popliteal fossa, including the tibial, peroneal, and sural nerves, also were injected with the HRP cocktail and their projections compared with those from the gastrocnemius muscle. Projections from the gastrocnemius muscle only partially overlapped with those from the tibial nerve, from which the nerves to the gastrocnemius muscle branch. However, the topology of projections from these nerves to laminae II-IV of the dorsal horn differed from that of the nerves of the gastrocnemius muscle, suggesting there was little spread to other nerves in the popliteal fossa. It was also noted that large labeled processes, presumably dendrites of retrogradely labeled motoneurons, entered the dorsal horn. These data provide information on the central projections of both the large and small fibers innervating the gastrocnemius muscle, and may aid in determining the circuitry utilized in the exercise pressor reflex as well as muscle pain.

Negative chronotropism of the heart is inhibited with lesions of the caudal medulla in the rat.

Neurons in the ventrolateral medulla are essential for cardiorespiratory regulation. It has been suggested that neurons in the caudal ventrolateral medulla are responsible for the negative chronotropic effect of the heart, at least in carnivores, because injection of glutamate into this area decreases heart rate significantly. In the present study, we monitored heart rate both before and after injections of the excitotoxin ibotenic acid into the most caudal part of the ventrolateral medulla in rats. We found that resting heart rate increased significantly by more than 53% (P<0.0001) after the ibotenic acid injections. This result suggests that neurons located in the caudal ventrolateral medulla are responsible for the negative chronotropic effect of the heart in the rat, especially its most caudal part.

Trigemino-autonomic connections in the muskrat: the neural substrate for the diving response.

Stimulation of the anterior ethmoidal nerve of the muskrat produces a cardiorespiratory depression similar to the diving response. This includes an apnea, a parasympathetic bradycardia, and a selective increase in sympathetic vascular tone. However, the brainstem circuitry that links the afferent stimulus to the efferent autonomic responses is unknown. We used the anterograde transneuronal transport of the herpes simplex virus (HSV-1), strain 129, after its injection into the anterior ethmoidal nerve to determine the primary, secondary, and tertiary brainstem relays responsible for this cardiorespiratory response. In an effort to check the validity of this relatively untested tracer, we also injected the medullary dorsal horn with biotinylated dextran amine to determine the secondary trigemino-autonomic projections. Approximately 1 microl (6x10(6) PFU) of the HSV-1 virus was injected directly into the anterior ethmoidal nerve of muskrats. After 2-6 days, their trigeminal ganglions, spinal cords and brainstems were cut and immunohistologically processed for HSV-1. Initially (2 days), HSV-1 was observed only in the trigeminal ganglion. After approximately 3 days, HSV-1 was observed first in many brainstem areas optimally labeled between 4 and 4.5 days. In these cases, the ventrolateral superficial medullary dorsal horn, the ventral paratrigeminal nucleus and the interface between the interpolar and caudal subnuclei were labeled ipsilaterally. The nucleus tractus solitarius (NTS), especially its ventrolateral, dorsolateral, and commissural subnuclei were labeled as well as the caudal, intermediate and rostral ventrolateral medulla. Within the pons, the superior salivatory nucleus, the A5 area, the ventrolateral part of the parabrachial nucleus and the Kölliker-Fuse nucleus were labeled. Only after a survival of 4 days or more, the locus coeruleus, the nucleus raphe magnus, the nucleus paragigantocellularis, pars alpha, and the pontine raphe nucleus were labeled. Injections of biotinylated dextran amine were made into the medullary dorsal horn (MDH) in a location similar to that labeled after the viral injections. Fine fibers and terminals were labeled in the same brainstem areas labeled after injections of HSV-1 into the anterior ethmoidal nerve. This study outlines the potential brainstem circuit for the diving response, the most powerful autonomic reflex known. It also confirms the efficacy for using HSV-1, strain 129, as an anterograde transneuronal transport method.

Electrical stimulation of the anterior ethmoidal nerve produces the diving response.

Stimulation of the upper respiratory tract usually produces apnea, but it can also produce a vagally mediated bradycardia and a sympathetically mediated increase in peripheral vascular resistance. This cardiorespiratory response, often called the diving response, is usually initiated by nasal stimulation. The purpose of this research was to investigate the anterior ethmoidal nerve (AEN) that innervates the nasal mucosa of muskrats (Ondatra zibethicus). Electrical stimulation of the AEN (typically 50 Hz, 100 micros and 500 microA) produced immediate and sustained bradycardia and cessation of respiration similar to that of the diving response. Heart rate (HR) significantly decreased from 264+/-18 to 121+/-8 bpm, with a concurrent 4.2+/-0.9 s apnea, during the 5 s stimulation period. BP decreased from 97.9+/-4.8 to 91.2+/-6.4 mmHg. Using estimations from (1) cross-sectional areas of AEN trigeminal ganglion cells labeled with WGA-HRP, and (2) electron microscopic analysis of the AEN, we found that approximately 65% of the AEN is composed of unmyelinated C-fibers. In addition, 72.4% of myelinated fibers from the nerves that innervate the nasal passages were of small diameter (<6 microm, presumably Adelta fibers). Thus, the AEN of the muskrat contains a high concentration of small diameter fibers (89.8%). We conclude that electrical stimulation of small diameter fibers within the AEN of muskrats can produce the cardiovascular and respiratory responses similar to that of the diving response.

The rostral ventrolateral medulla mediates the sympathoactivation produced by chemical stimulation of the rat nasal mucosa.

1. We sought to outline the brainstem circuit responsible for the increase in sympathetic tone caused by chemical stimulation of the nasal passages with ammonia vapour. Experiments were performed in alpha-chloralose-anaesthetized, paralysed and artificially ventilated rats. 2. Stimulation of the nasal mucosa increased splanchnic sympathetic nerve discharge (SND), elevated arterial blood pressure (ABP), raised heart rate slightly and inhibited phrenic nerve discharge. 3. Bilateral injections of the broad-spectrum excitatory amino acid receptor antagonist kynurenate (Kyn) into the rostral part of the ventrolateral medulla (RVLM; rostral C1 area) greatly reduced the effects of nasal mucosa stimulation on SND (-80 %). These injections had no effect on resting ABP, resting SND or the sympathetic baroreflex. 4. Bilateral injections of Kyn into the ventrolateral medulla at the level of the obex (caudal C1 area) or into the nucleus tractus solitarii (NTS) greatly attenuated the baroreflex and significantly increased the baseline levels of both SND and ABP. However they did not reduce the effect of nasal mucosa stimulation on SND. 5. Single-unit recordings were made from 39 putative sympathoexcitatory neurons within the rostral C1 area. Most neurons (24 of 39) were activated by nasal mucosa stimulation (+65.8 % rise in discharge rate). Responding neurons had a wide range of conduction velocities and included slow-conducting neurons identified previously as C1 cells. The remaining putative sympathoexcitatory neurons were either unaffected (n = 8 neurons) or inhibited (n = 7) during nasal stimulation. We also recorded from ten respiratory-related neurons, all of which were silenced by nasal stimulation. 6. In conclusion, the sympathoexcitatory response to nasal stimulation is largely due to activation of bulbospinal presympathetic neurons within the RVLM. We suggest that these neurons receive convergent and directionally opposite polysynaptic inputs from arterial baroreceptors and trigeminal afferents. These inputs are integrated within the rostral C1 area as opposed to the NTS or the caudal C1 area.

Evaluation of benzalkonium chloride chemoneurolytic proximal gastric vagotomy.

BACKGROUND: Transmucosal chemoneurolytic injection of benzalkonium chloride (BAC) has previously been shown to duplicate operative proximal gastric vagotomy (PGV) in controlling gastric acid secretion. In this study, BAC was evaluated as to efficacious dose, methods of delivery, and systemic toxicities. METHODS: Sham celiotomy, operative PGV controls, transmucosal injections through a gastrotomy, and transserosal injections of BAC (saline controls, 0. 625, 1.25, 2.5, 5.0, 10 mg BAC/kg body wt) were administered to Sprague-Dawley rats. After 3 months the rats underwent Congo red testing (CRT), horseradish peroxidase (HRP) neuronal staining, and necropsy. The color density change of the gastric mucosa from basic to acidic demonstrated by the CRT at the time of necropsy was used to calculate the residual anatomic acid-secreting area. Prior to necropsy, subserosal HRP injections into the anterior and posterior stomach walls assayed vagal neuronal viability via retrograde axonal flow. Results were compared by an ANOVA. RESULTS: The results demonstrated that 1.25-10 mg/kg transmucosal BAC replicated the results of operative PGV; 2.5 mg/kg was found to be the most effective dose. All injection groups including saline controls demonstrated similar diminished vagal retrograde axonal flow by HRP testing consistent with local BAC chemoneurolytic effects. No systemic toxic symptoms were observed after tail vein intravenous BAC 1.25, 2.5, and 5.0 mg/kg. CONCLUSIONS: These efficacy studies have demonstrated BAC's potential utility in the performance of endoscopic transmucosal chemoneurolytic PGV.

Fos immunohistochemical determination of brainstem neuronal activation in the muskrat after nasal stimulation.

Stimulation of the nasal passages of muskrats with either ammonia vapours or retrogradely-flowing water produced cardiorespiratory responses (an immediate 62% decrease in heart rate, 29% increase in mean arterial blood pressure, and sustained expiratory apnoea). We used the immunohistological detection of Fos, the protein product of the c-fos gene, as a marker of neuronal activation to help elucidate the brainstem circuitry of this cardiorespiratory response. After repeated ammonia stimulation of the nasal passages, increased Fos expression was detected within the spinal trigeminal nucleus (ventral laminae I and II of the medullary dorsal horn, ventral paratrigeminal nucleus, and spinal trigeminal nucleus interpolaris), an area just ventromedial to the medullary dorsal horn, the caudal dorsal reticular formation and the area of the A5 catecholamine group compared to control animals. Repeated water stimulation of the nasal passages produced increased Fos expression only in the A5 catecholamine group. There was an increase in the number of Fos-positive cells in the ammonia group in the ventral laminae I and II of the medullary dorsal horn and the ventral paratrigeminal nuclei compared with the water group. We conclude that ammonia stimulation of the nasal passages produces a different pattern of neuronal activation within the brainstem compared with water stimulation. We also conclude that Fos immunohistochemistry is a good technique to determine functional afferent somatotopy, but that immunohistochemical detection of Fos is not a good technique to identify the medullary neurons responsible for the efferent aspects of an intermittently produced cardiorespiratory reflex.

Brainstem origin of preganglionic cardiac motoneurons in the muskrat.

The muskrat, and aquatic rodent with a brisk and reliable diving response, shows a remarkable bradycardia after nasal stimulation. However, the medullary origin of cardiac preganglionic motoneurons is unknown in this species. We injected fat pads near the base of the heart of muskrats with a WGA-HRP solution to label retrogradely preganglionic parasympathetic neurons that project to the cardiac plexi. Results showed that the preponderance of labeled neurons was in ventrolateral parts of the medulla from 1.5 mm caudal to the obex to 2.0 mm rostral. Eighty-nine percent of the labeled neurons were located bilaterally in the external formation of the nucleus ambiguus, 5.6% were in the lateral extreme of the dorsal motor nucleus of the vagus nerve and 5.3% were found in the intermediate area in between these two nuclei. Although controversy still exists concerning the medullary origin of preganglionic cardiac motoneurons, our results from muskrats agree with those from most other species where preganglionic cardiac motoneurons were located just ventral to the nucleus ambiguus.

Trigeminally-mediated alteration of cardiorespiratory rhythms during nasal application of carbon dioxide in the rat.

Stimulation of the upper respiratory tract with air-borne irritants can result in dramatic alterations of cardiorespiratory rhythms that include apnea, bradycardia and selective peripheral vasoconstriction. Since carbon dioxide can stimulate receptors in the nasal passages, we wanted to determine if this odorless gas can induce the same autonomic changes as air-borne irritants. Passing 100% carbon dioxide through the nasal passages of rats anesthetized with chloralose-urethane produced apnea, a vagally-mediated bradycardia and a sympathetically-mediated increase in mean arterial blood pressure. Application of atropine blocked the bradycardia without affecting respiratory or blood pressure changes, while injection of prazosin eliminated blood pressure responses but did not affect heart rate or apnea. There were no significant autonomic responses to nasal application of 10, 25 or 50% carbon dioxide. The responses were mediated through the trigeminal innervation of the nasal mucosa since they could be blocked when the anesthetic procaine was applied to the nasal cavity. We conclude that these cardiorespiratory responses are due to stimulation of trigeminal nociceptors located within the nasal mucosa.

A medullary dorsal horn relay for the cardiorespiratory responses evoked by stimulation of the nasal mucosa in the muskrat Ondatra zibethicus: evidence for excitatory amino acid transmission.

Stimulation of the upper respiratory tract, including the nasal mucosa, with water, vaporous irritants, or gases, induces a collation of several cardiorespiratory responses including an apnea and bradycardia and often some change in arterial blood pressure. Since the nasal mucosa is innervated by branches of the trigeminal nerve, it implies that some part of the trigeminal system within the central nervous system mediates the autonomic responses induced by nasal stimulation. In the present study, respirations, heart rate and arterial pressure were monitored in muskrats anesthetized with a mixture of chloralose-urethane. We induced a bradycardia and apnea by stimulating the nasal mucosa of muskrats with brief (5 s) transnasal application of vapors of ammonia hydroxide. In an effort to determine the central site where the trigeminal mediation of the cardiorespiratory responses occurs, small nanoliter injections of 2% lidocaine were made bilaterally into the subnucleus caudalis of the spinal trigeminal nucleus (referred to as the medullary dorsal horn) to determine if the responses could be blocked. The responses could be blocked when the lidocaine injections on both sides were placed in the rostral, ventral parts of the medullary dorsal horn, but persisted when the injections were placed elsewhere. Since lidocaine blocks both neurons and fibers of passage, nanoliter injections of kynurenate, a general excitatory amino acid antagonist, were used in a similar paradigm to circumvent the problem of blocking only fibers of passage.(ABSTRACT TRUNCATED AT 250 WORDS)

Evaluation of alternative proximal gastric vagotomy techniques after a 9-month interval in a rat model.

Proximal gastric vagotomy (PGV) is an accepted operation for patients with ulcers that are refractory to medical management. Results comparable to those of standard, operative PGV have previously been demonstrated using endoscopic chemoneurolytic injection or laparoscopic laser seromyotomy in a porcine model. In this study, we evaluated several PGV techniques in regard to long-term effects on acid secretion, ulcer prophylaxis, and permanent vagal denervation in a rat model. Trans-mucosal injection of chemoneurolytic agents (cobaltous chloride, benzalkonium chloride, and phenol) and seromyotomy by CO2 laser were performed. After 9 months, all rats received sub-serosal gastric injections of horseradish peroxidase (HRP) during laparotomy. Twenty-four hours later, an ulcerogenic dose of pentagastrin was administered sub-cutaneously. Three days after administration of HRP (to allow time for retrograde axonal transport and labeling of cells of the dorsal vagal nucleus with HRP), necropsy was performed. The pre-pyloric gastric mucosa was inspected for ulcerogenic changes, and a Congo red solution was applied to the gastric mucosa to map the acid-secreting areas. All PGV methods significantly diminished pentagastrin-induced ulceration when compared to sham controls. Benzalkonium chloride chemoneurolytic and laser methods were most effective for decreasing the size of acid-secreting areas. A reduced number of HRP-stained cells in the dorsal vagal nucleus indicated permanent denervation of vagal-gastric connections by operative and laser techniques.

Trigeminal projections to the peribrachial region in the muskrat.

The anterograde and retrograde transport of wheat germ agglutinin-horseradish peroxidase was used to study the trigeminoperibrachial pathway in the muskrat after injections of tracer into either the medullary dorsal horn or the dorsolateral pons. After injections into the medullary dorsal horn, labeled fibers ascended into the ipsilateral dorsolateral pons via the spinal trigeminal tract, within the neuropil of the trigeminal sensory complex and within the reticular formation adjacent to the spinal trigeminal nucleus. At caudal levels of the ipsilateral peribrachial area, dense terminal-like label distributed in the Kölliker-Fuse nucleus continued into the lateral parabrachial nucleus. At intermediate levels ipsilaterally, the Kölliker-Fuse nucleus again was labeled densely, as were areas analogous to the external lateral and external medial subnuclei of the parabrachial nucleus in the rat. A thin band of label along the ventral spinocerebellar tract outlined an unlabeled area in the central portion of the lateral parabrachial nucleus. Rostrally near the pontomesencephalic junction, the area designated the superior lateral subnucleus in the hamster was labeled, while sparser label was present more dorsally. Contralateral to the injections, caudal and intermediate levels of the peribrachial area contained only scant reaction product. However, the rostral area of the superior lateral subnucleus was labeled densely via fibers ascending in the trigeminothalamic tract. Injections made just rostral to the obex and either centered in or including the dorsal or ventral paratrigeminal nuclei produced similar labeling at caudal and intermediate levels of the peribrachial area. An exception, however, was that the caudal medial parabrachial nucleus was also labeled after the dorsal paratrigeminal injection. Also, only scant label was found in the rostral third of the dorsolateral pons on either side after these injections. Both trigeminothalamic and trigeminolemniscal pathways were labeled contralaterally after these injections. These trigeminal projections to the dorsolateral pons were compared to the projections from the nucleus tractus solitarii and the ventrolateral medulla. Numerous trigeminal neurons were labeled retrogradely after injections of wheat germ agglutinin-horseradish peroxidase into the dorsolateral pons. In the medullary dorsal horn, they were found almost exclusively in laminae I and V. Labeled neurons in lamina I were especially prominent in rostral ventral levels of the medullary dorsal horn. Labeled cells in lamina I were continuous with others found in the displaced band of substantia gelatinosa at the interface of the subnucleus caudalis and subnucleus interpolaris, as well as with those found in the ventral and dorsal paratrigeminal nuclei.(ABSTRACT TRUNCATED AT 400 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)

Proximal gastric vagotomy by minimally invasive methods in an acute rat model.

In this prospective study, minimally invasive methods of proximal gastric vagotomy (PGV) were investigated in male Sprague-Dawley rats. Completeness of vagotomy by traditional operative therapy, by laser denervation of the gastric serosa, and by subserosal or transmucosal injections of chemoneurolytic agents was evaluated with postoperative Congo red testing, ulcerogenic stimulation of the gastric mucosa, and histochemical labeling of whatever vagal fibers remained in the gastric wall. Short-term results demonstrate that successful PGV can be performed with minimally invasive methods.

Trigeminal mediation of the diving response in the muskrat.

Stimulation of the nasal cavity elicits powerful cardiorespiratory responses similar to the diving response. In the present study, bradycardia and apnea were elicited in muskrats by stimulation of the nasal cavity with ammonia vapors. These responses could be blocked by injections of 2% lidocaine made bilaterally into the medullary dorsal horns of the trigeminal sensory complex. However, the bradycardia due to activation of the baroreceptor reflex with intravenous phenylephrine was retained. These data implicate trigeminal neurons in the medullary dorsal horn as modulators of autonomic activity, especially in the cardiorespiratory adjustments after nasal stimulation.

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 Kölliker-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)

Stereotaxic atlas of the brainstem of the muskrat, Ondatra zibethicus.

The muskrat is an aquatic rodent of moderate size and possesses dramatic respiratory and cardiovascular behaviors in response to submersion, stimulation of the upper respiratory tract, or environmental cues. This report provides a stereotaxic atlas of the muskrat's brainstem in the transverse plane to facilitate studies requiring an accurate localization of discrete brain structures. Plates are presented of brainstem sections between the obex and superior colliculus separated by 450 microns. Each figure shows a Nissl stained hemisection complemented by a line drawing. Our data shows mature muskrats have relatively uniform craniometric dimensions promoting their use for stereotaxic placements in their brains.

Trigeminal projections to contralateral dorsal horn originate in midline hairy skin.

The present study tested the hypothesis that the trigeminal (V) primary afferent projection to the contralateral dorsal horn originates in midline hairy skin. A prior study (Jacquin et al., 1990) showed that this crossed projection is heaviest to ophthalmic regions of medullary and cervical dorsal horns, and that it does not arise from V ganglion cells that innervate cornea, nasal mucosa, or cerebral dura mater. Here, retrograde double-labeling methods were used to show that many ophthalmic ganglion cells that innervate midline hairy skin via the supraorbital nerve project to the contralateral medullary and upper cervical dorsal horns. Diamidino yellow injections into the right dorsal horn labeled an average of 104 cells in the left V ganglion. Of these contralaterally projecting ganglion cells, an average of 45% were also labeled by horseradish peroxidase (HRP) injections into the left supraorbital nerve, and 25% were also labeled by HRP injections into the midline opthalmic hairy skin. However, only 2% were labeled by HRP injections restricted to left supraorbital vibrissae follicle nerves. Almost all of the double-labeled cells were located in the dorsal one-half of the V ganglion, and they did not differ in size from single-labeled cells. On the basis of these and prior data, we conclude that a high percentage of contralaterally projecting V ganglion cells originate in midline hairy skin. It is also likely that the contralaterally projecting V ganglion cells serve a low-threshold mechanoreceptive function, given the relatively large ganglion cells and axons giving rise to this pathway and their central terminations in dorsal horn laminae III-V.

Controlled bradycardia induced by nasal stimulation in the muskrat, Ondatra zibethicus.

Respiration was disrupted and bradycardia induced in anesthetized muskrats by stimulating the nasal cavity with a stream of either water or various concentrations of ammonia vapors. When responses induced by either ammonia or water were compared, ammonia vapors were considered preferable because the responses could be maintained reliably through relatively rapid periods of stimulation, and the post-stimulus recovery of heart rate and respiration was more predictable. Moreover, the bradycardia induced in the first 5 s of stimulation by dilutions of ammonia vapors was graded. After injections of lidocaine were made into the nucleus tractus solitarius a profound bradycardia to ammonia stimulation persisted despite disruption of normal respiratory rhythms and an inhibition of the baroreceptor reflex induced by phenylephrine administration. These results show that ammonia vapors stimulating the nasal chambers effectively elicit cardio-respiratory adjustments in anesthetized muskrats and that the bradycardia may be controlled by varying the intensity of the peripheral stimulus. The trigeminal contribution for this is emphasized since the bradycardia persists after reversible blockade of the solitary complex. These data suggest that the trigeminal input to cardiac motorneurons is via relatively few synapses and is over circuits which run parallel to those modulating cardiac activity in response to chemoreceptors, baroreceptors and pulmonary afferent fibers.