Degeneration of ingestion-related brainstem nuclei in spinocerebellar ataxia type 2, 3, 6 and 7

Dysphagia, which can lead to nutritional deficiencies, weight loss and dehydration, represents a risk factor for aspiration pneumonia. Although clinical studies have reported the occurrence of dysphagia in patients with spinocerebellar ataxia type 2 (SCA2), type 3 (SCA3), type 6 (SCA6) and type 7 (SCA7), there are neither detailed clinical records concerning the kind of ingestive malfunctions which contribute to dysphagia nor systematic pathoanatomical studies of brainstem regions involved in the ingestive process. In the present study we performed a systematic post mortem study on thick serial tissue sections through the ingestion-related brainstem nuclei of 12 dysphagic patients who suffered from clinically diagnosed and genetically confirmed spinocerebellar ataxias assigned to the CAG-repeat or polyglutamine diseases (two SCA2, seven SCA3, one SCA6 and two SCA7 patients) and evaluated their medical records. Upon pathoanatomical examination in all of the SCA2, SCA3, SCA6 and SCA7 patients, a widespread neurodegeneration of the brainstem nuclei involved in the ingestive process was found. The clinical records revealed that all of the SCA patients were diagnosed with progressive dysphagia and showed dysfunctions detrimental to the preparatory phase of the ingestive process, as well as the lingual, pharyngeal and oesophageal phases of swallowing. The vast majority of the SCA patients suffered from aspiration pneumonia, which was the most frequent cause of death in our sample. The findings of the present study suggest (i) that dysphagia in SCA2, SCA3, SCA6 and SCA7 patients may be associated with widespread neurodegeneration of ingestion-related brainstem nuclei; (ii) that dysphagic SCA2, SCA3, SCA6 and SCA7 patients may suffer from dysfunctions detrimental to all phases of the ingestive process; and (iii) that rehabilitative swallow therapy which takes specific functional consequences of the underlying brainstem lesions into account might be helpful in preventing aspiration pneumonia, weight loss and dehydration in SCA2, SCA3, SCA6 and SCA7 patients.     

Differential activation of medullary vagal nuclei during different phases of swallowing in the cat

The aim of this study was to identify the medullary vagal nuclei involved in the different phases of swallowing activated physiologically in a species with an esophagus similar to human. In decerebrate cats, the pharyngeal (0.5-1.0 ml water in pharynx (N=6)) or esophageal (1-3 ml air in esophagus (N=5)) phases of swallowing were stimulated separately once per minute for 3 h, and we compared the resulting c-fos immunoreactivity within neuronal cell nuclei of the dorsal motor nucleus (DMN), nucleus tractus solitarius (NTS) and nucleus ambiguus (NA) with a sham control group (N=5). We found that the pharyngeal phase was associated with an elevated number of c-fos positive neurons in the intermediate (NTSim), interstitial (NTSis), ventromedial (NTSvm) subnuclei of the NTS, caudal DMN, and dorsal NA; and the esophageal phase was associated with an elevated number of c-fos positive neurons in the central (NTSce), ventral, dorsolateral, ventrolateral subnuclei of the NTS, rostral DMN, and ventral NA. We concluded that the pharyngeal and esophageal phases of swallowing are associated with different sets of NTS subnuculei; and the DMN and NA may contain functionally different populations of motor neurons situated rostrocaudally and dorsoventrally associated with the different phases of swallowing. The central pattern generator (CPG) for swallowing probably receives significant peripheral feedback, and the NTSvm may participate in the transition of the pharyngeal to the esophageal phase of swallowing.     

Functional organization of vagal reflex systems in the brain stem of the goldfish, Carassius auratus.

The coordination of secretory and motor responses to food within the alimentary canal requires well organized brain stem reflex systems. In the goldfish, Carassius auratus, three vagal reflex systems control three phases of ingestion and digestion. The orobranchial system sorts food from substrate, the pharyngeal chewing organ prepares items deemed to be food for digestion and absorption, and the abdominal system regulates the digestion of food. Each system is represented in the central nervous system by separate sensory and motor nuclei. The aim of the present study was to determine whether the interrelationships among the vagal sensory and motor nuclei reflect the peripheral organization. The sensory nucleus of each vagal system was injected with the neuronal tracer horseradish peroxidase (HRP), in separate cases. HRP injections into the vagal lobe sensory layers (orobranchial system) labeled fibers projecting topographically to the vagal lobe motor layer, but not at all to the pharyngeal or abdominal motor nuclei. Similarly, injections of HRP into the pharyngeal and abdominal sensory nuclei selectively labeled nerve fibers projecting to the pharyngeal and abdominal motor nuclei, respectively. All injections resulted in labeled fibers and/or cells in the lateral reticular formation, and in fibers ascending in the secondary gustatory-visceral tract. Gustatory information from the pharynx is apparently processed in the same brain stem system as pharyngeal general visceral information, suggesting that functional or regional characteristics of visceral sensory information may be more important for brain stem processing than the traditional "special" (gustatory) versus "general" visceral dichotomy. These results indicate that anatomically and functionally separate reflex systems exist within the goldfish vagal visceral nuclei.     

Ingestion-related activity of caudate and entopeduncular neurons in the cat

Single-unit activity was recorded in the caudate nucleus and entopeduncular nucleus in cats during ingestion. In both nuclei, ingestion-related neurons were of two types: those with no facial somatosensory receptive field and an ingestion-related response, and those with a facial somatosensory field and an ingestion response. Neurons in both regions contained cells with conditioned responses to the noise produced by the solenoid that delivered milk. No caudate neurons and only 3% of entopeduncular neurons were phasically related to jaw muscle activity. These results indicate that neurons in both input and output regions of the basal ganglia in the cat have complex ingestion-related response characteristics with little relationship to specific orolingual movements.     

Axonal projections of medullary swallowing neurons in guinea pigs

A central pattern generator (CPG) for swallowing in the medulla oblongata generates spatially and temporally coordinated movements of the upper airway and alimentary tract. To reveal the medullary neuronal network of the swallowing CPG, we examined the cytoarchitecture of the swallowing CPG and axonal projections of its individual neurons by extracellular recording and juxtacellular labeling of swallowing-related neurons (SRNs) in the medulla in urethane-anesthetized and paralyzed guinea pigs. Three major types of neuronal discharge patterns were identified during fictive swallowing induced by stimulation of the superior laryngeal nerve: early (burst-like activation during the pharyngeal stage), late (activation after the pharyngeal stage), and inhibited (inhibition during the pharyngeal stage) types. Sixteen neurons were successfully labeled in the nucleus tractus solitarii (NTS) and in the medullary reticular formation (RF). No motoneuron was labeled. The SRNs in the NTS had axons projecting to the NTS, RF, nucleus ambiguus, nucleus hypoglossus, and dorsal motor nucleus of the vagus on the ipsilateral side. Some NTS SRNs projected only within the NTS. The axons of SRNs in the RF projected also to the NTS, RF, motor nuclei on the ipsilateral side, and to the other side RF. These findings show anatomic substrates for the neuronal network of the CPG for swallowing, which consists of complex neuronal connections among SRNs in the NTS, RF, and motor nuclei.     

Neurophysiology of swallowing.

Swallowing is a complex motor event that is difficult to investigate in man by neurophysiological experiments. For this reason, the characteristics of the brain stem pathways have been studied in experimental animals. However, the sequential and orderly activation of the swallowing muscles with the monitoring of the laryngeal excursion can be recorded during deglutition. Although influenced by the sensory and cortical inputs, the sequential muscle activation does not alter from the perioral muscles caudally to the cricopharyngeal sphincter muscle. This is one evidence for the existence of the central pattern generator for human swallowing. The brain stem swallowing network includes the nucleus tractus solitarius and nucleus ambiguus with the reticular formation linking synaptically to cranial motoneuron pools bilaterally. Under normal function, the brain stem swallowing network receives descending inputs from the cerebral cortex. The cortex may trigger deglutition and modulate the brain stem sequential activity. The voluntarily initiated pharyngeal swallow involves several cortical and subcortical pathways. The interactions of regions above the brain stem and the brain stem swallowing network is, at present, not fully understood, particularly in humans. Functional neuroimaging methods were recently introduced into the human swallowing research. It has been shown that volitional swallowing is represented in the multiple cortical regions bilaterally but asymmetrically. Cortical organisation of swallowing can be continuously changed by the continual modulatory ascending sensory input with descending motor output. Significance: Dysphagia is a severe symptom complex that can be life threatening in a considerable number of patients. Three-fourths of oropharyngeal dysphagia is caused by neurological diseases. Thus, the responsibility of the clinical neurologist and neurophysiologist in the care for the dysphagic patients is twofold. First, we should be more acquainted with the physiology of swallowing and its disorders, in order to care for the dysphagic patients successfully. Second, we need to evaluate the dysphagic problems objectively using practical electromyography methods for the patients' management. Cortical and subcortical functional imaging studies are also important to accumulate more data in order to get more information and in turn to develop new and effective treatment strategies for dysphagic patients.     

Distribution of choline acetyltransferase (ChAT) immunoreactivity in the brain of the adult trout and tract-tracing observations on the connections of the nuclei of the isthmus

The distribution of cholinergic neurons and fibers was studied in the brain and rostral spinal cord of the brown trout and rainbow trout by using an antiserum against the enzyme choline acetyltransferase (ChAT). Cholinergic neurons were observed in the ventral telencephalon, preoptic region, habenula, thalamus, hypothalamus, magnocellular superficial pretectal nucleus, optic tectum, isthmus, cranial nerve motor nuclei, and spinal cord. In addition, new cholinergic groups were detected in the vascular organ of the lamina terminalis, the parvocellular and magnocellular parts of the preoptic nucleus, the anterior tuberal nucleus, and a mesencephalic tegmental nucleus. The presence of ChAT in the magnocellular neurosecretory system of trout suggests that acetylcholine is involved in control of hormone release by neurosecretory terminals. In order to characterize the several cholinergic nuclei observed in the isthmus of trout, their projections were studied by application of 1,1;-dioctadecyl-3,3,3;, 3;-tetramethylindocarbocyanine perchlorate (DiI) to selected structures of the brain. The secondary gustatory nucleus projected mainly to the lateral hypothalamic lobes, whereas the nucleus isthmi projected to the optic tectum and parvocellular superficial pretectal nucleus, as previously described in other teleost groups. In addition, other isthmic cholinergic nuclei of trout may be homologs of the mesopontine system of mammals. We conclude that the cholinergic systems of teleosts show many primitive features that have been preserved during evolution, together with characteristics exclusive to the group.     

Videofluorographic study of swallowing in patients with severe motor and intellectual disabilities--II. Impairment of oral phase

We studied swallowing movements in 14 patients with severe motor and intellectual disabilities and development of the impaired oral phase. The oral phase in the patients did not develop beyond the late weaning phase. They had impairment of the oral phase: poor or absent lip seal, infantile swallowing, slowed or stalled bolus transit, and oral residue after swallowing. Videofluorographic studies revealed severe impairment in the transitional and pharyngeal phases: pooling of liquid barium in the hypopharynx prior to swallowing, the delay of cricopharyngeal relaxation, aspirated liquids (silent aspiration 77.8%), aspirated purees, pooling of purees in the pharynx after swallowing. After videofluorographic studies we made some efforts to provide patients with better management; changes in food textures and posture during drinking and eating. These efforts eased their feeding difficulties. Videofluorographic studies could be helpful in evaluating swallowing disturbance in the oral phase and thereby preventing chronic aspiration, malnutrition and feeding difficulties in patients with severe motor and intellectual disabilities.     

Upper airway motor outputs during vomiting versus swallowing in the decerebrate cat

Swallowing and vomiting are antagonistic motor acts; nevertheless, vomiting can be immediately followed by swallowing. The purpose of this study was to clarify the interrelationship between these two behaviors, particularly in regard to comparing the upper airway motor patterns at the end of the expulsion phase with those during subsequent swallowing. Experiments were conducted using both paralyzed and non-paralyzed decerebrate cats, in which recordings were obtained either from upper airway muscles, the diaphragm and abdominal muscles or from the nerves that innervate those muscles. The activity patterns of most nerves recorded in paralyzed animals were consistent with the behavior recorded in non-paralyzed animals from the muscles innervated by those nerves, with the exception of the cricothyroid and stylopharyngeus muscles. Vomiting can be divided into a series of retches followed by expulsion, which itself can be further subdivided into three phases. The final stage of expulsion, characterized by burst-like exaggerated activity of the laryngeal elevator thyrohyoid and the pharyngeal constrictors, proved to be different from pharyngeal swallowing, as judged from differences in the spatio-temporal patterns of the upper airway motor outputs. However, post-vomiting swallowing activity was still observed even after total deafferentation of the laryngeal and pharyngeal areas in paralyzed animals. It is therefore likely that the central processes for vomiting and swallowing closely relate in generating these two behaviors.     

Task-dependent differences in corticobulbar excitability of the submental motor projections: Implications for neural control of swallowing

It has been suggested that the primary motor cortex plays a substantial role in the neural circuitry that controls swallowing. Although its role in the voluntary oral phase of swallowing is undisputed, its precise role in motor control of the more reflexive, pharyngeal phase of swallowing is unclear. The contribution of the primary motor cortex to the pharyngeal phase of swallowing was examined using transcranial magnetic stimulation (TMS) to evoke motor evoked potentials (MEPs) in the anterior hyomandibular muscle group during either volitional submental muscle contraction or contraction during the pharyngeal phase of both volitionally, and reflexively, initiated swallowing. For each subject, in all three conditions, TMS was triggered when submental surface EMG (sEMG) reached 75% of the mean maximal submental sEMG amplitude measured during 10 volitional swallows. MEPs recorded during volitional submental muscle contraction were elicited in 22 of the 35 healthy subjects examined (63%). Only 16 of these 22 subjects (45.7%) also displayed MEPs recorded during volitional swallowing, but their MEP amplitudes were larger when triggered by submental muscle contraction than when triggered by volitional swallowing. Additionally, only 7 subjects (of 19 tested) showed MEPs triggered by submental muscle contraction during a reflexively triggered pharyngeal swallow. These differences indicate differing levels of net M1 excitability during execution of the investigated tasks, possibly brought about by task-dependent changes in the balance of excitatory and inhibitory neural activity.     

Organization of some brain stem afferents to the paraventricular nucleus of the hypothalamus in the rat

Brain stem afferents to subnuclei of the paraventricular nucleus of the hypothalamus were studied by the anterograde autoradiographic technique in the rat. The parabrachial nuclei and locus coeruleus project to the posterior, periventricular, parvocellular and dorsal divisions. The ventral medulla projects to the posterior, medial, lateral, parvocellular and dorsal divisions. The A1 catecholamine cell group projects to the posterior, medial, lateral, parvocellular and dorsal divisions, and the nucleus of the solitary tract to the parvocellular and dorsal divisions. The A1 region and the ventral medulla also project to the supraoptic nucleus.     

Morphological characteristics of acetylcholinesterase-containing neurons in the CNS of DFP-treated monkeys. Part 3. Brain stem and spinal cord.

The distribution of acetylcholinesterase (AChE, EC 3.1.1.7) was studied in the lower brain stem and spinal cord of 4 monkeys following the i.m. administration of bis-(1-methylethyl) phosphorofluoridate (di-isoprophylfluorophosphate: DFP). In 1 animal that received 0.43 mg/kg of DFP 4 hr prior to death, AChE was virtually absent in all structures. In the other 3 animals sacrificed 10, 12 and 18 hr after the injection of 0.20 mg/kg of DFP, AChE activity was considerably lighter in the neuropil of different structures normally displaying a moderate to intense AChE activity in pharmacologically unmanipulated monkeys. As a consequence of the lighter background activity several groups of neurons were easily identified and their cell bodies and processes were sharply outlined. Brain stem and spinal cord groups of neurons that show an intense AChE activity include the nuclei of the somatic motor column (cranial nerves III, IV, VI and XII, and ventral horn cells) and of the special visceral (cranial nerves V and VII and nucleus ambiguus) and general visceral (Edinger-Westphal and salivatory nuclei, dorsal nucleus of the Xth nerve and intermediomedial and intermediolateral spinal nuclei) motor columns. Other neurons of the midbrain that display intense AChE activity include the rostral division of nucleus linearis, the magnocellular division of the red nucleus, perirubral giant neurons, the nucleus of the mesencephalic root of V, the substantia nigra and the subnucleus compactus of the pedunculopontine tegmental nucleus. Midbrain neurons with a light to moderate AChE activity are located in the periaqueductal gray, the parvocellular division of the red nucleus, the interstitial nucleus of Cajal, and the magnocellular nucleus of the posterior commissure. Other intensely stained groups of neurons at isthmus and pontine levels include the intermediate and caudal divisions of nucleus linearis, all divisions of the dorsal nucleus of the raphé, the laterodorsal nucleus, nucleus annularis, nucleus centralis superior, neurons of the loci coeruleus and subcoeruleus and of the mesencephalic root of V, the few and large neurons of nuclei pontis oralis and pontis caudalis and nuclei paraabducens and paramedianus dorsalis. Other groups of neurons with a light to moderate AChE activity at isthmus and pons levels include the pontine and reticulotegmental nuclei. The neurons of the cerebellar fastigial nucleus are intensely stained and those of nucleus interpositus and nucleus dentatus, as well as the cell bodies of the Golgi cells of the cerebellar cortex, are moderately stained. At medulla and spinal cord levels the neurons of the lateral vestibular nucleus, the gigantocellular nucleus, the dorsal nucleus of Clarke and the lumbo-sacral border cells are intensely stained. Other neurons with lightly to moderately stained cell bodies include the superior and medial vestibular nuclei, nucleus praepositus, the lateral cuneate and lateral reticular nuclei, and the principal inferior and accessory olivary nuclei.     

Neuronal analysis of pharyngeal peristalsis in the gastropod Navanax in terms of identified motoneurons innervating identified muscle bands. II. Radial and circumferential motor fields

The neuronal basis of pharyngeal ingestion and peristalsis was studied in the gastropod Navanax inermis. Radially and circumferentially oriented muscles produced expansion and constriction of the pharynx. Motor fields of 11 identified radial motoneurons and 13 identified circumferential motoneurons were determined with respect to circumferential and longitudinal muscle band coordinates by muscle movements, electromyography, antidromic stimulation and axonal anatomy. Activation of these identified motoneurons can account for all the elemental pharyngeal movements observed during feeding. Four motoneurons, each innervating most of radial muscle, can mediate ingestion. Three radial motoneurons with anterior motor fields can mediate anterior expansion during sealing of the pharyngeal lips around prey and during regurgitation. Ten circumferential motoneurons have small arciform motor fields, the distributions of which correspond to the regional specializations in circumferential band organization. Arciform constriction can center eccentric ingested prey within the pharyngeal lumen during peristalsis. Arciform constrictions could combine to form an annular constriction in peristalsis. Small, non-overlapping, circumferential motor fields maximize the number of independent annular units available to produce a fine peristaltic wave. Sphincters have more circumferential motoneurons with smaller motor fields; this innervation permits finer motor control. Radial motoneurons with posterior motor fields can produce expansion caudal to a circumferential constriction during peristalsis. Motor fields of regional radial motoneurons show greater interanimal variability than circumferential motor fields, which is correlated with a less essential role of radial motoneurons in peristalsis. Two circumferential motoneurons with giant posterior pharyngeal motor fields can mediate pharyngeal emptying either in swallowing or in regurgitation.     

The projections of the dorsomedial hypothalamic nucleus in the rat

The dorsomedial hypothalamic nucleus (DMH) output pathways are revealed by using autoradiographic tracing of tritium labeled Leucine and by the recently introduced Phaseolus vulgaris leuco-agglutinin immunocytochemical method. Terminal labeling appears in the dorsal motor nucleus of the vagus, nucleus ambiguus and in the parvocellular reticular formation at the lower medullary level. Mesencephalic labeling is found in the periaqueductal gray at the level of the oculomotor nucleus. In the hypothalamus labeled terminal boutons are identified in the lateral and ventromedial hypothalamic nuclei but also in the parvocellular paraventricular nucleus. Furthermore, the circumventricular organs are found to receive a dense DMH input, particularly the organum vasculosum of the lamina terminalis and the subfornical organ. These findings are discussed in relation to the dorsomedial nucleus involvement in the control of feeding and pancreatic hormone release. It appears that the DMH participates in this control via descending pathways to the preganglionic pancreas innervating neurons but also via a neuroendocrine route. The latter connection is indicated by terminal labeling in the parvocellular paraventricular nucleus in the area that contains the corticotropin-releasing factor positive cells.     

Distribution of vasoactive intestinal polypeptide (VIP) in the rat brain stem nuclei

The distribution of vasoactive intestinal polypeptide (VIP) in the individual nuclei of the rat brain stem was examined. Highest VIP concentrations in brain stem (> 1 ng/mg protein) are confined to dorsal regions, including periaqueductal gray and structures under the fourth ventricle. VIP concentrations are moderately high (0.8–1 ng/mg protein) in gracile nucleus, area postrema, nucleus of the solitary tract, motor nucleus of the XIIth, locus ceruleus and dorsal tegmental nucleus. Cerebellum and pontine nuclei contained only very low levels (< 0.2 ng/mg protein) of VIP.     

Neuronal activity in the medulla oblongata during vocalization. A single-unit recording study in the squirrel monkey

In six squirrel monkeys (Saimiri sciureus), the medulla oblongata was explored with microelectrodes, looking for vocalization-correlated activity. The vocalizations were elicited by microinjections of glutamate agonists into the periaqueductal grey of the midbrain. Vocalization-related cells were found in greater numbers in the nucl. ambiguus (Ab) and retroambiguus (RAb), in the parvocellular, magnocellular and central reticular formation as well as in the solitary tract nucleus and spinal trigeminal nucleus. Small numbers were also found in the vestibular complex, cuneate nuclei, inferior olive and lateral reticular nucleus. A differentiation of the neuronal responses into 12 reaction types reveals that the frequency of each reaction type varies from brain structure to brain structure, thus allowing a specification of the different vocalization-related areas. According to this specification, it is proposed that initiation of vocalization takes place via the parvocellular reticular formation; vocal pattern control is mainly brought about by the parvocellular reticular formation, Ab, solitary tract nucleus and spinal trigeminal nucleus; expiratory control and respiratory-laryngeal coordination is carried out by the RAb, Ab and central nucleus of the reticular formation; vocalization-specific postural adjustments are carried out via the vestibular and cuneate nuclei.     

Neuronal analysis of pharyngeal peristalsis in the gastropod Navanax in terms of identified motoneurons innervating identified muscle bands. II. Radial and circumferential motor fields

The neuronal basis of pharyngeal ingestion and peristalsis was studied in the gastropod Navanax inermis. Radially and circumferentially oriented muscles produce expansion and constriction of the pharynx. Motor fields of 11 identified radial motoneurons and 13 identified circumferential motoneurons were determined with respect to circumferential and longitudinal muscle band coordinates by muscle movements, electromyography, antidromic stimulation and axonal anatomy. Activation of these identified motoneurons can account for all the elemental pharyngeal movements observed during feeding. Four motoneurons, each innervating most of radial muscle, can mediate ingestion. Three radial motoneurons with anterior motor fields can mediate anterior expansion during sealing of the pharyngeal lips around prey and during regurgitation. Ten circumferential motoneurons have small arciform motor fields, the distributions of which correspond to the regional specializations in circumferential band organization. Arciform constriction can center eccentric ingested prey within the pharyngeal lumen during peristalsis. Arciform constrictions could combine to form an annular constriction in peristalsis. Small, non-overlapping, circumferential motor fields maximize the number of independent annular units available to produce a fine peristaltic wave. Sphincters have more circumferential motoneurons with smaller motor fields; this innervation permits finer motor control. Radial motoneurons with posterior motor fields can produce expansion caudal to a circumferential constriction during peristalsis. Motor fields of regional radial motoneurons show greater interanimal variability than circumferential motor fields, which is correlated with a less essential role of radial motoneurons in peristalsis. Two circumferential motoneurons with giant posterior pharyngeal motor fields can mediate pharyngeal emptying either in swallowing or in regurgitation.     

Immunocytochemical localization of the choline acetyltransferase containing neuron system in the rat lower brain stem

This distribution of choline acetyltransferase (CHAT) immunoreactivity (CHAT-I) in the rat lower brain stem was analyzed using a highly sensitive avidin-biotin immunocytochemical method and 3-amino-9-ethyl-carbazole visualization. A much wider and more abundant distribution of CHAT-I structures in the lower brain stem was demonstrated than in earlier studies. The following areas were newly identified as areas rich in CHAT-I fibers: the interpeduncular nucleus, medial geniculate body, central gray matter of pons, pontine nucleus, parabigeminal nucleus, dorsal tegmental nucleus of Gudden, lateral trapezoid nucleus, inferior colliculus, dorsal and ventral cochlear nuclei, medial and lateral vestibular nuclei, reticular formation of medulla oblongata, and gelatinosa of caudal trigeminal spinal tract nucleus. In addition to the areas in which they have been known to exist, CHAT-I perikarya were found in the caudal portion of substantia nigra pars reticulata, the area between trigeminal motor nucleus and superior olivary nucleus, the medial and spinal vestibular nucleus, prepositus hypoglossal nucleus, raphe magnus and obscurus, ventromedial portion of solitary tract nucleus and its just ventral reticular formation, and caudal trigeminal spinal tract nucleus.