Observations on the afferent and efferent organization of the vagus nerve and the innervation of the stomach in the squirrel monkey

Horseradish peroxidase was injected into the cervical vagus nerve or stomach wall of adult squirrel monkeys. Following cervical vagus nerve injections, labelled afferent fibres were present in the tractus solitarius and labelled fibres and terminals were present in medial and lateral parts of the nucleus of the tractus solitarius (NTS) ipsilaterally. Afferent labelling was also seen in the ipsilateral commissural nucleus and in the area postrema. Labelling was present contralaterally in caudal levels of the medial parts of the NTS, in the commissural nucleus, and in the area postrema. Afferent projections to the ipsilateral pars interpolaris of the spinal trigeminal nucleus and to the substantia gelatinosa of the C1 segment of the spinal cord were also labelled. Following injections of HRP into the anterior and posterior stomach walls, the tractus solitarius was labelled bilaterally. Afferent labelling was concentrated bilaterally in the dorsal parts of the medial division of the NTS, i.e., in the subnucleus gelatinosus, and in the commissural nucleus. The regions of NTS immediately adjacent to the tractus solitarius were largely unlabelled. Injections of HRP into the cervical vagus nerve resulted in heavy retrograde labelling of neurons in the ipsilateral dorsal nucleus of the vagus (DMX) and in the nucleus ambiguus (NA). In addition a few neurones were labelled in the intermediate zone between these two nuclei. Retrogradely labelled neurons were also present in the nucleus dorsomedialis in the rostral cervical spinal cord and in the spinal nucleus of the accessory nerve. Injections of HRP into the left cricothyroid muscle in two cases resulted in heavy retrograde labelling of large neurons in the left NA. Following stomach wall injections of HRP retrograde labelling of neurons was seen throughout the rostrocaudal and mediolateral extent of the DMX; there was no apparent topographical organization of the projection. In these cases, a group of labelled smaller neurons was found lying ventrolateral to the main part of the NA through its rostral levels. This study in a primate indicates that a large vagal afferent projection originates in the stomach wall and terminates primarily in the subnucleus gelatinosus of the NTS and in the commissural nucleus with a distribution similar to that described previously in studies in several subprimate mammalian species. The present results and those of other studies suggest some degree of segregation of visceral input within different subnuclei of the NTS.(ABSTRACT TRUNCATED AT 400 WORDS)     

Vagal and gastric connections to the central nervous system determined by the transport of horseradish peroxidase

Horseradish peroxidase (HRP, Sigma Type VI) crystals were encased in a parafilm envelope and applied to the transected central ends of the left and right cervical vagus nerves and the anterior and posterior esophageal vagus nerves of adult male hooded rats. Injections of 30% HRP were made into the muscle wall of the fundus and antrum regions of the stomach. After 48 hr survival time, animals were perfused intracardially with a phosphate buffer plus sucrose wash followed by glutaraldehyde and paraformaldehyde fixative. The brain stem, spinal cord and corresponding dorsal root ganglia, superior cervical sympathetic ganglion, and the nodose ganglion were removed and cut into 50 micron sections. All tissue was processed with tetramethylbenzidine (TMB) for the blue reaction according to Mesulum and counterstained with neutral red. Sequential sections were examined under a microscope. Labeled neurons and nerve terminals were identified using bright and dark field condensers and polarized light. In tissue from animals that had HRP applied to the cervical vagus nerves, retrogradely labeled neurons were identified ipsilaterally in the medulla located in the dorsal motor nucleus of the vagus (DMN) and the nucleus ambiguus (NA). Labeled cells extended from the DMN into the spinal cord in ventral-medial and laminae X regions C1 and C2 of cervical segments. Many neurons were labeled in the nodose ganglion. Anterogradely labeled terminals were observed throughout and adjacent to the solitary nucleus (NTS) dorsal to the DMN and intermixed among labeled neurons located in the DMN. In tissue from animals that had HRP applied to the esophageal vagus nerves, similar labeling was observed. However, fewer neurons were identified in the NA, the nodose ganglion, and only in laminae X of the cervical spinal cord segments C1 and C2. Also, very little terminal labeling was observed in and adjacent to the NTS. Labeled neurons in tissue from animals that had HRP injected into the stomach wall were observed bilaterally in the DMN, nodose ganglion, and only in laminae X at the C1 and C2 levels of the spinal cord. Labeled neurons also were observed in the dorsal root ganglia of the thoracic cord. These data indicate that cervical cord and NA neurons are important in the supradiaphragmatic motor innervation by the vagus. Also, many afferents to the NTS originate above the diaphragm. In addition, some afferents from the stomach enter the central nervous system via the thoracic spinal cord.     

Central distribution of the cervical vagus nerve in Old and New World primates

The central distribution of the cervical vagus nerve was examined in Old and New World primates using anterograde transganglionic and retrograde horseradish peroxide (HRP) histochemistry. Crystals of HRP were applied to the cut central end of the cervical vagus nerve in two Old World (one bonnet, one cynomolgus) and two New World (squirrel) monkeys. Bright- and darkfield examination of coronal sections from the pons, medulla, and upper cervical spinal cord revealed two major concentrations of retrogradely labeled cells in the ipsilateral dorsal motor nucleus (DMX) and nucleus ambiguous (NA). DMX was heavily labeled, containing about 5 times as many labeled cells as NA. The anterograde distribution of reaction product did not extend as far in the rostrocaudal plane as did the retrograde distribution. Labeled afferent fibers entered the medulla at the level of the caudal dorsal cochlear nucleus, joined the solitary tract, and descended to the obex. Ipsilateral terminal label first appeared at the level where the nucleus of the solitary tract (NST) abuts the IVth ventricle. The terminal field grew in extent and density, until at the level of the area postrema (AP), the distribution extended throughout the medial NST, ventrolateral NST, and AP. Contralateral terminal label was sparse and restricted to the medial NST. In the commissural division of the solitary nucleus, sparse reaction product was present bilaterally, with the denser concentration ipsilateral to the treated nerve. Examination of peripheral ganglia revealed labeled somata in the nodose, jugular, and superior cervical ganglia.     

The location of extrinsic efferent and afferent nerve cell bodies of the normal canine stomach

The location of the extrinsic efferent and afferent nerve cell bodies to the mucosa, submucosa, and tunica muscularis of the cardiac, gastric, and pyloric gland regions of the ventral stomach and to the mucosa-submucosa alone of these 3 glandular gastric regions was determined using the horseradish peroxidase technique. All animals of the study demonstrated labeling bilaterally in the rostrocaudal extent of the dorsal motor nucleus of the vagus nerve (DMV) although mucosa-submucosa injections resulted in fewer labeled cells in the DMV. There was no evidence of viscerotopic organization within the DMV for the different gastric regions. However, the left nucleus generally contained a greater number of labeled cells than the right nucleus. Injection of the mucosa, submucosa, and tunica muscularis of the cardiac gland region also resulted in labeling in the nucleus ambiguus in 4 of 5 animals. The vast majority of labeled postganglionic sympathetic neurons were found in the celiacomesenteric ganglion. Labeled cells were also located variously in the stellate ganglion, middle cervical ganglion, and sympathetic trunk ganglia for the different groups. There was no discernible pattern of localization of labeled cells within a sympathetic ganglion. For the stomach, afferent labeled cells were located in the range of the first thoracic to fourth lumbar spinal ganglia and the nodose ganglia, bilaterally. As with sympathetic neurons, there was no discernible pattern of localization of labeled cells within a sensory ganglion.     

Brainstem distribution of neurons with efferent projections in the cervical vagus of the dog

The distribution within the brainstem of cell bodies and efferent fibers projecting in the cervical vagus was studied with retrograde transport of horseradish peroxidase (HRP). Five to eight days after multiple microinjections of HRP into either the cervical vagosympathetic trunk or the nodose ganglion the brainstems and nodose ganglia were perfused and processed by the tetramethyl benzidine method. HRP-positive neurons were found in three brainstem regions: a dorsal cell column comprising the dorsal motor nucleus of the vagus (dmnX), a ventrolateral group in the region of nucleus ambiguus (nA), and scattered cells along a line between these columns. The density of labeled neurons was greatest within dmnX. Axons from cells of the ventrolateral column projected dorsomedially; just ventral to dmnX they turned laterally to exit the medulla in multiple rootlets. Within nA labelled neurons were distributed according to size, with larger cells more medial and smaller ones more lateral. Caudal to nA in nucleus retroambigualis and nucleus dorsalis medialis cell bodies appeared segregated into clusters.     

Afferent projections of the cervical vagus and nodose ganglion in the dog

The distribution within the brain stem of the afferent projections of the cervical vagus and the nodose ganglion was studied with horseradish peroxidase (HRP) and HRP-wheat germ agglutinin conjugate. Two to eight days after application of tracer into the cervical vagosympathetic trunk or the nodose ganglion the brain stems and ganglia were perfused and processed by the tetramethyl benzidine method. Vagal afferent fibers entered the lateral medulla as a distinct bundle spatially separate from the vagal efferent rootlets which were caudal and ventral to the afferents. Labeled axons in the solitary tract began to enter the nucleus tractus solitarii (nTS) 4.5 mm anterior to obex and were seen throughout the ipsilateral nTS as far as 3.5 mm caudal to obex. Label density varied within the nTS, with heaviest labeling in the dorsal and dorsolateral portions. Label was also seen in the ipsilateral area postrema (ap) and dorsal motor nucleus of the vagus. Labeled fibers crossed in the commissural portions of ap and nTS to enter the contralateral ap and nTS.     

Brain stem projections of sensory and motor components of the vagus complex in the cat: I. The cervical vagus and nodose ganglion

The motor and sensory connections of the cervical vagus nerve and of its inferior ganglion (nodose ganglion) have been traced in the medulla oblongata of 32 adult cats with the neuroanatomical methods of horseradish peroxidase (HRP) histochemistry and amino acid autoradiography (ARG). In 14 of these subjects, an aqueous solution of HRP was applied unilaterally to the central end of the severed cervical vagus nerve. In 13 other cases, HRP was injected directly into the nodose ganglion. Three of these 13 subjects had undergone infranodose vagotomy 6 weeks prior to the HRP injection. A mixture of tritiated amino acid was injected into the nodose ganglion in five additional cats. The retrograde transport of HRP yielded reaction product in nerve fibers and perikarya of parasympathetic and somatic motoneurons in the medulla oblongata. Furthermore, a tetramethyl benzidine (TMB) method for visualizing HRP enabled the demonstration of anterograde and transganglionic transport, so that central sensory connections of the nodose ganglion and of the vagus nerve could also be traced. The central distribution of silver grain following injections of tritiated amino acids in the nodose ganglion corresponded closely with the distribution of sensory projections demonstrated with HRP, thus confirming the validity of HRP histochemistry as a method for tracing these projections. The histochemical and autoradiographic experiments showed that the vagus nerve enters the medulla from its lateral aspect in multiple fascicles and that it contains three major components—axons of preganglionic parasympathetic neurones, axons of skeletal motoneurons, and central processes of the sensory neurons in the nodose ganglion. Retrogradely labeled neurons were seen in the dorsal motor nucleus of X(dmnX), the nucleus ambiguus (nA), the nucleus retroambigualis (nRA), the nucleus dorsomedialis (ndm) and the spinal nucleus of the accessory nerve (nspA). The axons arising from motoneurons in the nA did not traverse the medulla directly laterally; rather, all of these axons were initially directed dorsomedially toward the dmnX, where they formed a hairpin loop and then accompanied the axons of dmnX neurons to their points of exit. Afferent fibers in the vagus nerve reached most of the subnuclei of the nTS bilaterally, with the more intense labeling being found on the ipsilateral side. Labeling of sensory vagal projections was also found in the area postrema of both sides and around neurons of the dmnX. These direct sensory projections terminating within the dmnX may provide an anatomical substrate for vagally mediated monosynpatic reflexes. Following deefferentiation by infranodose vagotomy 6 weeks prior to HRP injections into the nodose ganglion, a number of neurons in the dmnX were still intensely labeled with the HRP reaction product. The axons of these HRP-labeled perikarya may constitute the bulbar component of the accessory nerve.     

Projections from the nucleus tractus solitarii to the rostral ventrolateral medulla

Projections from the nucleus tractus solitarii (NTS) to autonomic control regions of the ventrolateral medulla, particularly the nucleus reticularis rostroventrolateralis (RVL), which serves as a tonic vasomotor center, were analyzed in rat by anterograde, retrograde, and combined axonal transport techniques. Autonomic portions of the NTS, including its commissural, dorsal, intermediate, interstitial, ventral, and ventrolateral subnuclei directly project to RVL as well as to other regions of the ventrolateral medulla. The projections are organized topographically. Rostrally, a small cluster of neurons in the intermediate third of NTS, the subnucleus centralis, and neurons in proximity to the solitary tract selectively innervate neurons in the retrofacial nucleus and nucleus ambiguus. Neurons generally located in more caudal and lateral sites in the NTS innervate the caudal ventrolateral medulla (CVL). The RVL, CVL, and nucleus retroambiguus are interconnected. A combined retrograde and anterograde transport technique was developed so as to prove that projections from the NTS to the ventrolateral medulla specifically innervate the region of RVL containing neurons projecting to the thoracic spinal cord or the region of the nucleus containing vagal preganglionic neurons. When the retrograde tracer, fast blue, was injected into the thoracic spinal cord, and wheat germ agglutinin-conjugate horseradish peroxidase (HRP) was injected into the NTS, anterogradely labeled terminals from the NTS surrounded the retrogradely labeled neurons in the RVL and in the nucleus retroambiguus in the caudal medulla. Among the bulbospinal neurons in the RVL innervated by the NTS were adrenaline-synthesizing neurons of the C1 group. When fast blue was applied to the cervical vagus, and HRP was injected into the NTS, anterogradely labeled terminals from the NTS surrounded retrogradely labeled neurons in the rostral dorsal motor nucleus of the vagus, the region of the nucleus ambiguus, the retrofacial nucleus, and the dorsal portion of the RVL, a region previously shown to contain cardiac vagal preganglionic neurons. This combined anterograde and retrograde transport technique provides a useful method for tracing disynaptic connections in the brain. These data suggest that the RVL is part of a complex of visceral output regions in the ventrolateral medulla, all of which receive afferent projections from autonomic portions of the NTS. Bulbospinal neurons in the RVL, in particular the C1 adrenaline neurons, may provide a portion of the anatomic substrate of the baroreceptor and other visceral reflexes.     

Representation of the cecum in the lateral dorsal motor nucleus of the vagus nerve and commissural subnucleus of the nucleus tractus solitarii in rat.

Motor fibers of the accessory celiac and celiac vagal branches are derived from the lateral columns of the dorsal motor nucleus of the vagus nerve. These branches also contain sensory fibers that terminate within the nucleus of the tractus solitarii. This study traces the innervation of the intestines by using the tracer cholera toxin-horseradish peroxidase. In 53 rats, the tracer was injected into either the stomach, duodenum, jejunum, terminal ileum, cecum, or ascending colon. With all cecal injections, prominent retrograde labeling of cell bodies occurred bilaterally in the lateral columns of the dorsal motor nucleus of the vagus nerve above, at, and below the level of the area postrema. Dendrites of laterally positioned neurons projected medially and rostrocaudally within the dorsal motor nucleus of the vagus nerve and dorsomedially into both the medial subnucleus and parts of the commissural subnucleus of the nucleus of the tractus solitarii. Sensory terminal labeling occurred in the dorsolateral commissural subnucleus at the level of the rostral area postrema and the medial commissural subnucleus caudal to the area postrema. Additionally, there was sensory terminal labeling within a small confined area of the dorsomedial zone of the nucleus of the tractus solitarii immediately adjacent to the fourth ventricle at a level just anterior to the area postrema. Stomach injections labeled motoneurons of the medial column of the entire rostrocaudal extent of the dorsal motor nucleus of the vagus nerve and a sensory terminal field primarily in the subnucleus gelatinosus, with less intense labeling extending caudally into the medial and ventral commissural subnuclei. Dendrites of gastric motoneurons project rostrocaudally and mediolaterally within the dorsal motor nucleus of the vagus nerve and dorsolaterally within the nucleus of the tractus solitarii. They are most pronounced at the level of the rostral area postrema where many dendrites course dorsolaterally terminating primarily within the subnucleus gelatinosus. Injections of the duodenum labeled a small number of the cells within the medial aspects of the dorsal motor nucleus of the vagus nerve. Jejunal, ileal, and ascending colon injections labeled cells sparsely within the lateral aspects of the dorsal motor nucleus of the vagus nerve bilaterally. No afferent terminal labeling was evident after injection of these areas of the bowel.(ABSTRACT TRUNCATED AT 400 WORDS)     

The motor nuclei and primary projections of the IXth,Xth, XIth,and XIIth cranial nerves in the monitor lizard,Varanus exanthematicus

The motor nuclei and sensory connections of the IXth, Xth, XIth, and XIIth cranial nerves of the reptile Varanus exanthematicus were studied with the methods of anterograde degeneration and anterograde and retrograde axonal transport. The motor nuclei of nerve IX are located ventrally in the rhombencephalon and are constituted medially by the large-celled glossopharyngeal part of the nucleus ambiguus and laterally by the small-celled nucleus salivatorius inferior. The motor nuclei of nerve X consist of the dorsomedially located dorsal motor nucleus of the vagus and the laterally located vagal part of the nucleus ambiguus. The rostral portion of the latter cell group contains smaller cells than its caudal portion and is rostrally continuous with the nucleus salivatorius inferior of nerve IX. The efferent axons of nerves IX and X arising from the ventrolateral medulla first course dorsomedially, form genua beneath the IVth ventricle, and then exit the brainstem. All primary afferent fibers of nerve IX and the majority of those of nerve X enter the solitary tract. Terminations of vagal fibers were observed in the postvagal portion of the nucleus of the solitary tract, the dorsal motor nucleus of the vagus, and the nucleus of the commissura infima. A small contingent of vagal fibers courses caudally just dorsolateral to the descending trigeminal tract. A separate spinal component of nerve XI could not be found. The bulbar component of this nerve forms part of nerve X and takes its main origin from a detached caudal element of the nucleus ambiguus. The motor nuclear complex of nerve XII consists of a large dorsal nucleus and a small ventral nucleus that extend from the medulla oblongata into the first segment of the cervical spinal cord.     

The central organization of the vagus nerve innervating the stomach of the rat

We employed the neural tracers cholera toxin-horseradish peroxidase and wheat germ agglutinin-horseradish peroxidase to examine the organization of the afferent and efferent connections of the stomach within the medulla oblongata of the rat. The major finding of this study is that gastric motoneurons of the dorsal motor nucleus (DMN) possess numerous dendrites penetrating discrete regions of the overlying nucleus of the solitary tract (NTS). In particular, dendritic labelling was present in areas of NTS which also received terminals of gastric vagal afferent fibers such as the subnucleus gelatinosus, nucleus commissuralis, and medial nucleus of NTS. This codistribution of afferent and efferent elements of the gastric vagus may provide loci for monosynaptic vagovagal interactions. A small number of dendrites of DMN neurons penetrated the ependyma of the fourth ventricle and a few others entered the ventral aspect of the area postrema, thus making possible the direct contact of preganglionic neurons with humoral input from the cerebrospinal fluid and/or the peripheral plasma. Nucleus ambiguus neurons projecting to the stomach predominantly innervate the forestomach. The dendrites of these cells, when labelled, were generally short, and extended beyond the compact cluster of ambiguus neurons in a ventrolateral direction, parallel to the fascicles of vagal efferent fibers traversing the medulla.     

Projection sites of superficial and deep spinal dorsal horn cells in the nucleus tractus solitarii of the rat

By using anterograde transport of biotin dextran amine injected into the cervical spinal dorsal horn, we have shown that fibres from superficial and deep dorsal horn project to the nucleus tractus solitarii via two distinct pathways. Afferent fibres from the superficial lamina (I-III) were found to course in the dorsal funiculus and terminate bilaterally in the caudal zone of the nucleus tractus solitarii (NTS), mainly within the commissural subnucleus. In contrast, afferents from the deeper dorsal horn laminae (IV-V) were found to course in the dorsolateral fasciculus and terminate ipsilaterally, mostly in the lateral areas of the caudal nucleus tractus solitarii. Similar, but more extensive patterns of labelled fibres were produced by injections into the white matter of the dorsal funiculus and dorsolateral fasciculus, respectively. These observations suggest that the caudal NTS not only serves as a location of visceral afferent convergence and integration, but may also be a receptive area for monosynaptic projections from dorsal horn neurons receiving sensory afferent inputs. Such projections may represent pathways through which NTS neurons are influenced by nociceptive and non-nociceptive information from the dorsal horn and thereby can co-ordinate the appropriate autonomic response, including adjustments in cardiorespiratory reflex output.     

Brainstem cells of origin of physiologically identified cardiopulmonary nerves in the rhesus monkey (Macaca mulatta)

The distributions of brainstem cells of origin of the cervical vagus nerve, its cervical and thoracic branches, and of neurons projecting to the cricothyroid muscle and the stomach wall were identified and compared following injections of horseradish peroxidase (HRP) in 18 Rhesus monkeys. Physiologically and/or anatomically identified cardiopulmonary nerves were injected with 3–20 μl of HRP to identify the locations of vagal preganglionic cardioinhibitory neurons in 10 of these monkeys. After injections into cardiopulmonary nerves, retrogradely labelled cells were concentrated ipsilaterally in the most lateral parts of the dorsal motor nucleus of the vagus nerve (DMV) and in the ventrolateral nucleus ambiguus (NA). Fewer labelled neurons were identified close to or in the principal (dorsal) division of the NA and in the intermediate zone between the DMV and NA. The results indicate that monkey cardiopulmonary nerves have multiple origins; their somata are located primarily in the ventrolateral NA and to a lesser extent in the lateral DMV. In monkeys, there is a stronger representation in the lateral DMV than in cat, dog and pig. The viscerotopic organization of the cells of origin of primate vagal nerves is similar to that in other species. The cells of origin of vagal projections to the superior laryngeal nerve and cricothyroid muscle were located in the NA rostrally to those of the inferior laryngeal nerve. Injections into the superior laryngeal nerve also resulted in significant labelling in the DMV and intermediate zone (IZ). The cells of origin of projections to the anterior stomach wall were restricted to the DMV with a bilateral distribution of labelled cells, concentrated medially in the nucleus.     

Cells of origin of motor axons in the subdiaphragmatic vagus of the rat

The central cell groups that give rise to the motor axons that travel in the subdiaphragmatic vagus were re-examined in the rat by transecting the dorsal or ventral vagus near the stomach and incubating the nerve stump in crystalline horseradish peroxidase (HRP). An exceedingly large percentage of cells was labeled throughout the dorsal motor nucleus of the vagus (mX), with labeled cells extending even beyond the rostro-caudal limits of the nucleus usually assigned on the basis of cytoarchitecture alone. Different patterns of cell-labeling could be correlated with one or the other of the two vagal branches. Incubation of the ventral branch labeled cells only in the left mX, while incubation of the dorsal branch labeled cells on both sides, although more extensively on the right. HRP-positive somata were also observed bilaterally in the nucleus ambiguus (NA) after incubation of either branch of the subdiaphragmatic vagus; this finding is in contrast to previous accounts in which motor fibers from NA were considered to project only to cervical and thoracic structures. These results suggest that mX and NA are responsible for a substantial component of abdominal innervation in the rat.     

The target specificity of the extrinsic innervation of the rat small intestine

The target specificity of the extrinsic innervation of the rat small intestine was examined by simultaneously injecting the proximal and distal small intestine with either wheat germ agglutinin-horseradish peroxidase (HRP) or fast blue. The number of single- and double-labeled cells in the nodose, dorsal root and coeliac-superior mesenteric ganglia and the dorsal motor nucleus of the vagus were counted and expressed as percentages of total labeled cells. Cells containing both HRP and Fast blue projected to both regions of the intestine. We found that the nodose and mesenteric ganglia contained significantly fewer double-labeled neurons (approximately 3 and 9% respectively) than the dorsal motor nucleus (19%) or dorsal root ganglion (20%). Presumably, a large number of double-labeled afferent or efferent neurons would limit the ability of a given component of the extrinsic innervation to control the activity of restricted regions of the small intestine (but might be important in overall regulation of intestinal function). In a separate series of experiments we examined the topography of neurons in the dorsal motor nucleus of the vagus labeled with HRP injection into either the proximal or distal small intestine. Both of these injections labeled neurons in the entire rostro-caudal extent of the nucleus, though approximately 75% of the cells were located between 720 microns caudal and 720 microns rostral to the obex. Cells in the rostral regions were found primarily in the lateral pole of the nucleus, whereas caudal regions contained labeled cells in both the medial and lateral poles.     

The origins of innervation of the canine caudal pharyngeal muscles: an HRP study

In deglutition, movements of the tongue and oropharynx direct a bolus to the laryngopharynx. The major muscles of this region, which includes the ‘cricopharyngeal sphincter’, must undergo sequential relaxation and contraction for correct swallowing action. The innervation of the caudal pharyngeal muscles involved in this action in the dog have not been determined previously by sensitive neuroanatomical techniques. In this study, the location of efferent and afferent neurons innervating the left cricopharyngeus, thyropharyngeus and hyopharyngeus muscles was determined by horseradish peroxidase (HRP) histochemistry in 7 puppies. Labeled cells were found ipsilaterally in the supraspinal nucleus, nucleus ambiguus (including nucleus retrofacialis) and nucleus intercalatus of all animals, in the parasympathetic nucleus of X (dorsal vagal efferent nucleus) of 6 animals, and in the hypoglossal nucleus of 4 animals. Small numbers of HRP-labeled cells were found contralaterally in the supraspinal nucleus of all animals, and in the rostral nucleus ambiguus, in the nucleus intercalatus and the parasympathetic nucleus of X of fewer animals. This defines a more extensive source of efferent neurons for these muscles than had been reported for the cat. Labeled postganglionic sympathetic neurons were found bilaterally in the cranial (superior) cervical, middle cervical and cervicothoracic (stellate) ganglia. Labeled afferent neurons were seen bilaterally in the proximal vagal (jugular) and distal vagal (nodose) ganglia and in the C1–C4 spinal ganglia. The location of sympathetic and sensory nerve cell bodies of the muscles of the laryngopharynx has not been previously reported.     

Subnuclear organization of the human caudal nucleus of the solitary tract

The caudal human nucleus of the solitary tract (NTS) is composed of 10 subnuclei. The commissural subnucleus spans the midline below the obex, merging rostrally into the medial subnucleus. The other subnuclei of the NTS are best seen just above the obex. The ventrolateral subnucleus contains large, darkly staining neurons. The interstitial subnucleus consists of neurons lying in groups intermingled with the fibers of the tract. The lateral subnucleus is small at caudal levels, merging with the interstitial subnucleus more rostrally. The dorsal subnucleus contains large melanotic neurons and encircles the substantia gelatinosus, a round, cell-poor subnucleus. The ventromedial subnucleus curls around the medial and ventral edge of the tract. The intermediate subnucleus, laying ventrolateral to the dorsal motor nucleus of the vagus, also contains melanotic neurons. The subpostremal subnucleus separates the area postrema from the NTS proper. The medial subnucleus is the largest subnucleus in the caudal NTS, containing medium-sized fusiform neurons. Adoption of a uniform cytoarchitectural map of the caudal NTS will permit more accurate comparisons between human and nonhuman studies.     

Association of neurotensin binding sites with sensory and visceromotor components of the vagus nerve

Specific neurotensin (NT) binding sites were recently shown to be highly concentrated in the nucleus of the solitary tract (NTS), which receives primary vagal afferents, and in the dorsal motor nucleus of the vagus (DMN), which contains the cell bodies of origin of vagal preganglionic neurons. To investigate the relationship of these binding sites with sensory and visceromotor components of the vagus nerve, they were labeled here in vitro, using monoiodo[Tyr3]neurotensin (125I-NT) and visualized by light microscopic radioautography in the dorsomedial medulla of both intact and unilaterally vagotomized rats, in the nodose ganglia of intact animals, and in ligated vagus nerves. Unilateral vagotomy performed above the nodose ganglion resulted in a significant ipsilateral decrease in 125I-NT binding within both the NTS and the DMN, suggesting that NT binding sites were associated with both primary afferent fibers and preganglionic nerve cell bodies. The selective radioautographic labeling of a subpopulation (approximately 15%) of neuronal perikarya in the nodose ganglion confirmed that a proportion of vagal afferent neurons contained NT binding sites. Following vagus nerve ligation, a pile up of radiolabeled NT binding sites was observed on both sides of the nerve crush, indicating that NT receptor components were transported both anterogradely and retrogradely along fibers of the vagus nerve. We conclude that NT receptors are synthesized and transported within a subpopulation of afferent and efferent components of the vagus nerve and that NT may therefore act presynaptically upon vagal axon terminals in both central and peripheral nervous systems.     

Brainstem cells of origin of the cervical vagus and cardiopulmonary nerves in the neonatal pig (Sus scrofa)

The distribution of the cells of origin of the cervical vagus and cardiopulmonary nerves has been studied in neonatal piglets (Sus scrofa) ranging in age from 1 to 60 days. Cardiopulmonary nerves were identified physiologically and anatomically prior to injection of horseradish peroxidase (HRP) into the nerves. Following injection of HRP into the cervical vagus nerve retrogradely labeled neurons were present in the dorsal motor nucleus of the vagus nerve (DMV), the nucleus of the solitary tract, the nucleus ambiguus (NA), ventrolateral to the NA and in an intermediate zone between the DMV and the NA. Two unique clusters of neurons were also retrogradely labeled after injections into the vagus nerve. One group was located lateral to the most caudal levels of the DMV and extended as far caudally as the C1 spinal segment. The second distinctive group was located ventrolateral to the nucleus ambiguus in a cell column identified as the ventrolateral nucleus ambiguus (VLNA). After injections of HRP into cardiopulmonary nerves, the majority of neurons were found in the VLNA and the distinct clusters of neurons in this cell column were particularly heavily labeled. Small numbers of cells were labeled in the DMV and NA and none were labeled in the solitary nucleus after cardiopulmonary nerve injections. There were no apparent age-related differences in the degree or distribution of retrograde labeling.The distribution of neurons in the medulla oblongata projecting into cardiopulmonary nerves in the piglet is similar to that described in other species, i.e., the nucleus ambiguus, particularly its ventrolateral cell column, is the primary site of cardiomotor neurons. In addition, in the piglet there is a morphologically distinct cluster of cells related to the heart, and possibly the lungs, which does not appear to be present in other species.