Spinothalamic tract neurons in the substantia gelatinosa.

The substantia gelatinosa of the mammalian spinal cord is generally believed to be a closed system; that is its neurons are thought to project only to the substantia gelatinosa of the same or the contralateral side. Experiments in monkeys, using injections of the marker enzyme horseradish peroxidase, show that at least some neurons of the substantia gelatinosa project to the thalamus and thus belong to the spinothalamic tract. Such neurons include two cell types intrinsic to the gelatinosa, the central cells and the limitrophe cells of Cajal.

Structural and functional changes in the cristae ampullares of aged gerbils.

We have reported that calretinin and calbindin staining of calyxes in the apical region of the cristae is reduced or absent in old gerbils (>or=35 months) that had normal numbers of hair cells [Kevetter GA, Leonard RB (2002) Decreased expression of calretinin and calbindin in the labyrinth of old gerbils. Brain Res 957:362-365]. Here we examine the ability of primary afferents in aged gerbils to carry a tracer injected into the vestibular nuclear complex to their terminals in the cristae. Calyxes throughout the cristae were well labeled in a young animal with such an injection. In the aged animals, many calyxes were only partially filled or not filled at all. In some cases labeled axons were also missing from the stroma underlying the missing calyxes. There is a strong correspondence between the region where the calyxes were not filled and the absence of calretinin immunostaining. To determine if afferents from the cristae are functionally abnormal, we recorded from their axons and attempted to activate them with natural stimulation. Among afferents that could be activated, we encountered many afferents that had spontaneous activity but could not be modulated with natural stimulation. When tested, the firing rate of these afferents could be modulated with galvanic stimulation, and/or they could be activated by pulsed electrical stimulation. We also encountered afferents that had no spontaneous activity. The presence of these axons was revealed by an injury discharge that could not be modulated with natural stimulation. When tested, these axons could be activated with pulsed electrical stimulation. In some instances we encountered two or more such afferents in a row, an event we have not seen in young animals. We suggest that the simplest explanation for these observations is that calyxes are being lost in old animals.

Muscarinic acetylcholine receptor subtype expression in avian vestibular hair cells, nerve terminals and ganglion cells.

Muscarinic acetylcholine receptors (mAChRs) are widely expressed in the CNS and peripheral nervous system and play an important role in modulating the cell activity and function. We have shown that the cholinergic agonist carbachol reduces the pigeon's inwardly rectifying potassium channel (pKir2.1) ionic currents in native vestibular hair cells. We have cloned and sequenced pigeon mAChR subtypes M2-M5 and we have studied the expression of all five mAChR subtypes (M1-M5) in the pigeon vestibular end organs (semicircular canal ampullary cristae and utricular maculae), vestibular nerve fibers and the vestibular (Scarpa's) ganglion using tissue immunohistochemistry (IH), dissociated single cell immunocytochemistry (IC) and Western blotting (WB). We found that vestibular hair cells, nerve fibers and ganglion cells each expressed all five (M1-M5) mAChR subtypes. Two of the three odd-numbered mAChRs (M1, M5) were present on the hair cell cilia, supporting cells and nerve terminals. And all three odd numbered mAChRs (M1, M3 and M5) were expressed on cuticular plates, myelin sheaths and Schwann cells. Even-numbered mAChRs were seen on the nerve terminals. M2 was also shown on the cuticular plates and supporting cells. Vestibular efferent fibers and terminals were not identified in our studies. Results from WB of the dissociated vestibular epithelia, nerve fibers and vestibular ganglia were consistent with the results from IH and IC. Our findings suggest that there is considerable co-expression of the subtypes on the neural elements of the labyrinth. Further electrophysiological and pharmacological studies should delineate the mechanisms of action of muscarinic acetylcholine receptors on structures in the labyrinth.

The organization of the extraocular motor nuclei in the Atlantic stingray, Dasyatis sabina.

Retrograde transport of HRP was used to determine the location and organization of the motor nuclei innervating the extrinsic eye muscles of the stingray, an elasmobranch fish. oculomotor neurons are located both medial to and immediately ventrolateral to the MLF in the rostral midbrain. A ventral oculomotor nucleus was found among the IIIrd nerve rootlets close to the base of the midbrain. The dendrites of cells in the dorsal nucleus appear to be preferentially oriented in the transverse plane penetrating the MLF. Motoneuron pools innervating individual muscles are incompletely segregated in the dorsal group. However, the ventral nucleus innervates only the inferior oblique muscle. Dorsally, motoneurons innervating a single muscle are found on both sides of the MLF. In the caudal midbrain, the majority of trochlear motoneurons are located immediately ventrolateral to the MLF. Abducens motoneurons are scattered in the medulla from a ventrolateral position resembling the location of the nucleus in teleost fish to a dorsomedial position close to the MLF as in most other vertebrates. In contrast to other vertebrates, the medial rectus muscle is innervated by the contralateral oculomotor nucleus. Motoneurons innervating the other muscles have the same laterality as found in other vertebrates.

Immunohistochemical studies on the distribution and origin of candidate peptidergic primary afferent neurotransmitters in the spinal cord of an elasmobranch fish, the Atlantic stingray (Dasyatis sabina).

The distribution and origin of four peptide neurotransmitter candidates of primary afferents (substance P, SP; somatostatin, SS; cholecystokinin, CCK; and vasoactive intestinal polypeptide, VIP) were studied in the stingray with peroxidase-antiperoxidase (PAP) immunohistochemistry. This elasmobranch has virtually no unmyelinated primary afferents, having instead only large and small myelinated afferents. SP-like immunoreactivity was distributed densely in the superficial aspect of the substantia gelatinosa (SG), particularly laterally, and was scattered in the nucleus proprius, the intermediate zone, and the ventral horn. The distributions of SS-, CCK-, and VIP-like immunoreactivities were similar to each other, but different from that of SP. Stained fibers appeared to issue from a prominent tract in the dorsolateral funiculus to form a plexus at the lateral margin of the nucleus proprius. The fibers spread dorsally and medially through the SG to terminate in a thin band at the superficial margin of the SG. Both SS and CCK were more dense in the lateral third of the SG, while VIP was more diffusely distributed within this structure. The remaining regions of the spinal gray matter contained immunoreactive fibers and terminals in variable densities. Many SS-positive cell bodies were observed in the ventral horn, in the deep dorsal horn, and in the ependymal layer. CCK-positive cells were observed in the medial ventral horn, and VIP-positive cells were observed subjacent to the SG and within the dorsolateral funiculus. After unilateral dorsal rhizotomies, SP-like immunoreactivity in the SG was depleted, while SP staining elsewhere and all SS, CCK, and VIP staining was indistinguishable from control. Thus all four peptides are present in the stingray spinal cord, although only SP appears to be a candidate primary afferent transmitter.

Use of calcium-binding proteins to map inputs in vestibular nuclei of the gerbil.

We wished to determine whether calbindin and/or calretinin are appropriate markers for vestibular afferents, a population of neurons in the vestibular nuclear complex, or cerebellar Purkinje inputs. To accomplish this goal, immunocytochemical staining was observed in gerbils after lesions of the vestibular nerve central to the ganglion, the cerebellum, or both. Eleven to fourteen days after recovery, the brain was processed for immunocytochemical identification of calretinin and calbindin. After lesion of the vestibular nerve, no calretinin staining was seen in any of the vestibular nuclei except for a population of intrinsic neurons, which showed no obvious change in number or staining pattern. Calbindin staining was reduced in all nuclei except the dorsal part of the lateral vestibular nuclei. The density of staining of each marker, measured in the magnocellular medial vestibular nucleus, was significantly reduced. After the cerebellar lesion, no differences in calretinin staining were noted. However, calbindin staining was greatly reduced in all nuclei. The density of staining, measured in the caudal medial vestibular nucleus, was significantly lower. After a combined lesion of the cerebellum and vestibular nerve, the distribution and density of calretinin staining resembled that after vestibular nerve section alone, whereas calbindin staining was no longer seen. This study demonstrates that calretinin and calbindin are effective markers for the identification of vestibular afferents.

Vestibulo-oculomotor connections in an elasmobranch fish, the Atlantic stingray, Dasyatis sabina.

In elasmobranch fishes, including the Atlantic stingray, the medial rectus muscle is innervated by the contralateral oculomotor nucleus. This is different from most vertebrates, in which the medial rectus is innervated by the ipsilateral oculomotor nucleus. This observation led to the prediction that the excitatory vestibulo-extraocular motoneuron projections connecting each semicircular canal to the appropriate muscle should use a contralateral projection from the vestibular nuclei to the motoneurons. This hypothesis was examined in the Atlantic stingray by injecting horseradish peroxidase unilaterally into the oculomotor nucleus. It was found that vestibulo-oculomotor projections arise from the ipsilateral anterior octaval nucleus and the contralateral descending octaval nucleus. The same pattern was observed when the trochlear nucleus was involved in the injection. There were no cells labeled in the region of the abducens nucleus, and no candidate for a nucleus prepositus hypoglossus was identified. The presence of compensatory eye movements, the directional sensitivity of the semicircular canals, the location of the motoneurons innervating each eye muscle, and our results indicate that the excitatory input to the extraocular motoneurons is derived from the contralateral descending octaval nucleus, and the inhibitory input is derived from the ipsilateral anterior octaval nucleus. The absence of both abducens internuclear interneurons and a nucleus prepositus hypoglossus suggests that eye movements, particularly those in the horizontal plane, are controlled differently in elasmobranchs than in other vertebrates examined to date.

The octavolateral systems in the stingray, Dasyatis sabina. I. Primary projections of the octaval and lateral line nerves.

The central projections of the electrosensory, mechanosensory, and octaval nerves of the Atlantic stingray were examined by transganglionic transport of horseradish peroxidase. Particular attention was paid to the relation of the projections to cell plates C1 and C2, and to a newly described cell plate, C3. The electroreceptors in the stingray are found in three groups on the dorsal and ventral sides of the body. The electroreceptors are represented topographically on the ipsilateral dorsal nucleus. Those of the rostral part of the head and pectoral fins are represented on the rostroventral part of the nucleus, and those on the caudal part of the head and the pectoral fin on the dorsocaudal part of the nucleus. Mechanosensory lateral line afferents terminate within the ipsilateral intermediate and caudal nuclei, and the lateral granular mass of the vestibulocerebellum. Anterior lateral line afferents also project to the magnocellular octaval nucleus. A topographic representation of the mechanosensory lateral line periphery is present on the intermediate and caudal nuclei. Mechanoreceptors on the trunk are represented laterally and those on the head medially. The terminal field of the anterior lateral line afferents on the intermediate nucleus surrounds cell plates C1 and C2. The anterior lateral line afferents also project to the medial part of the lateral granular mass, whereas the posterior lateral line afferents project to the lateral portion. Sparse projections of anterior lateral line afferents to the periventricular octaval nucleus were also observed. The octaval nerve afferents terminate largely within the octaval column. Octaval nerve projections were also observed to the reticular formation, the periventricular octaval nucleus, the deep cerebellar nucleus, the vestibulocerebellum, particularly the lower lip and medial granular mass, and the intermediate nucleus. A sparse projection to cell plate C3 was found. The relation between cell plates C1 and C2 and the anterior lateral line afferents suggests that these cell plates are related to processing lateral line information. While the relationship between cell plate C3 and the octaval afferents is not strong, the sparse octaval projection C3 receives suggests that it is relaying octaval information.

The organization of the electromotor nucleus and extraocular motor nuclei in the stargazer (Astroscopus y-graecum).

The organization of the oculomotor and electromotor systems was examined in the stargazer, a teleost. The electromotor system in these animals is a derivative of the oculomotor system. The extraocular motor nuclei and nerves consist of approximately equal numbers of motoneurons and axons (about 100 per muscle). In contrast, electromotor axons appear to branch several times within the intracranial portion of the IIIrd nerve. The topographical organization of the motoneurons was examined using retrograde transport of horseradish peroxidase injected into the electric organ or eye muscles. Electromotor and oculomotor neurons form distinct populations. Each electric organ receives a strong ipsilateral and a weak contralateral innervation. Individual eye muscles receive unilateral innervations with the expected laterality. Within the oculomotor nucleus there is some topographical separation of motoneurons innervating each muscle. Antidromic field potentials confirm the identity of the electromotor nucleus.

A cytoarchitectonic analysis of the spinal cord of the pigeon (Columba livia).

The spinal gray of the pigeon is described cytoarchitectonically to establish a foundation for anatomical and physiological studies of the pigeon spinal cord. The material includes segments from the high cervical cord through the lumbosacral enlargement, and nine cellular layers are described. In addition to this laminar organization, various distinct cell groups such as the dorsal magnocellular column, column of Terni, marginal cells and lobes of Lachi are described. Layers I-IV occupy the head of the dorsal horn, are apparent at all spinal levels examined, and represent the clearest case of laminar organization of the spinal gray of the pigeon. Layer V occupies the full extent of the neck of the dorsal horn at all segmental levels investigated. Also, the dorsal magnocellular column is situated in the central region of this layer from the rostral pole of the cervical enlargement through the lumbosacral enlargement, and arguments are advanced that this cell column is homologous to the column of Clarke. In the intermediate zone a Layer IV is defined, but it is apparent only at the enlargements. Layers VII-IX constitute the ventral horn, Layer IX being the motoneuronal cell groups. With the exception of the motoneuronal groups, the boundaries of the ventral horn layers are considerably less distinct than those of the dorsal horn, and no attempt is made to distinguish Layers VII and VIII at lumbosacral levels. At the enlargements there is a prominent lateral motoneuronal cell group consisting of large cells. It is generally concluded that the cytoarchitectonic organization of the spinal gray of the pigeon bears a rather close resemblance to that described for various mammalian species, particularly with respect to the dorsal horn.

Spinal terminal fields of dorsal root fibers in the pigeon (Columba livia).

This paper describes the spinal pattern of brachial and lumbosacral dorsal root terminations in the pigeon. These are presented within the framework of the cytoarchitectonic analysis of the preceding report (Leonard and Cohen, '75). At the level of dorsal root section and immediately adjacent sections dense terminal fields were evident throughout the ipsilateral dorsal horn (Layers I-V), including the substantia gelatinosa. However, degeneration in the substantia gelatinosa was prominent only with short survival times. There were mediolateral differences in the density of degenerating material in the dorsal horn, as well as variations between the patterns at the cervical and lumbosacral enlargements. The general pattern of degeneration in the dorsal horn extended approximately three segments rostral and two segments caudal to the level of root section, becoming progressively less dense with distance from the level of rhizotomy. An exception to this was degeneration in the dorsal magnocellular (Clarke's) column where, following section of either brachial or lumbosacral roots, degeneration could be traced into the thoracic spinal cord. However, these ascending and descending projections upon column at thoracic levels overlap minimally if at all. Degeneration was sparse in the intermediate zone and ventral horn (Layers VI-IX). It tended to concentrate centrally in Layer VI at the enlargements, and much of the degeneration ventral to this was interpreted as fibers en passage to the motoneuronal cell groups, Layer IX. Terminal fields in Layer IX were evident at both cervical and lumbosacral enlargements. They were largely restricted to the lateral motoneuronal cell group and were more prominent at cervical levels where degeneration was commonly observed around cell bodies and their proximal dendrites. As with the cytoarchitectonic organization of the spinal gray in the pigeon, the terminal fields of brachial and lumbosacral dorsal root fibers appear to have a pattern similar to that reported in mammalian literature.

Neuronal increase in various areas of the nervous system of the guppy, Lebistes.

The numbers of 1) dorsal root ganglion cells in the 2nd spinal segment, 2) ventral horn cells in the 2nd spinal segment, 3) Purkinje cells of the cerebellum, and 4) neurons in the nucleus glomerulosus were counted and correlated with age and size in the guppy, Lebistes. The findings were that the neuronal numbers in all these areas increased throughout much of the life of the animal. These data, combined with the previously demonstrated increases in retinal neurons in goldfish and sensory and spinal neurons in stingrays, suggest that neurons are added to many areas of the nervous system of fish as the animal ages and grows. In this respect, the nervous systems of fish differ from the nervous systems of other vertebrates. We offer the suggestion that the comparatively greater ability of fish to regenerate their nervous system after injury may be related in part to their ability to add neurons to various parts of the nervous system throughout life.

The cells of origin of the primate spinothalamic tract.

Spinothalamic tract cells in the lumbar, sacral and caudal segments of the primate spinal cord were labelled by the retrograde transport of horseradish peroxidase (HRP) injected into the thalamus. The laminar distribution of stained spinothalamic cells in the lumbosacral enlargement differed according to whether the HRP was injected into the lateral or the medial thalamus. Lateral injections labelled cells in most laminae, but the largest numbers of cells were in laminae I and V. The highest concentrations of cells labelled from the medial thalamus were in laminae VI-VIII. Ninety percent or more of the stained spinothalamic cells in the lumbosacral enlargement were contralateral to the injection site. In the conus medullaris stained spinothalamic cells were most numerous in laminae I, V and VI following lateral thalamic injections of HRP. Many of the cells of the conus were in Stilling's nucleus. Twenty-three percent of the cells in the conus were ipsilateral to the injection site in the lateral thalamus. Only a few cells in the conus were labelled by medial thalamic injections. The total number of spinothalamic cells from L5 caudally was estimated to be at least 1,200-2,500. An injection of HRP into the midbrain resulted in laminar distribution of labelled cells much like that produced by a lateral thalamic injection. The types of spinothalamic tract cells and the sizes of their somata were determined for different laminae. The cell types resemble those already described from Golgi and other studies of the spinal cord gray matter. The spinothalamic tract cells in lamina I included Waldeyer cells and numerous small fusiform, pyriform or triangular cells. Those in lamina II included limitrophe and central cells. Spinothalamic cells in lamina III were central cells. Most of the labelled cells in laminae IV-X were polygonal, although there were also flattened cells in these layers. The smallest spinothalamic cells were in laminae I-III, while the largest were in laminae V and VII-IX. Spinothalamic cells in the conus medullaris included cells like those in the lumbosacral enlargement, but also a special cell type in Stilling's nucleus. Some cells in the conus had dendrites that crossed the midline. Spinothalamic axons could sometimes be traced to the ventral white commissure within one or a few sections. In longitudinal sections, most labelled axons were in the ventral part of the lateral funiculus on the side of the injection, although a few were in the ventral funiculus or on the contralateral side. The axons were widely dispersed, and a few were located adjacent to the pia-glial membrane.

The descending and intrinsic serotoninergic innervation of an elasmobranch spinal cord.

The descending and the intrinsic components of the serotoninergic (5HT) innervation of the Atlantic stingray spinal cord were described by comparing the distributions of neuronal elements exhibiting 5HT-like immunoreactivity (peroxidase-antiperoxidase method) in sections caudal and rostral to spinal transections. The cells of origin of the descending 5HT system were located with a double labeling method for both retrogradely transported horseradish peroxidase (HRP) and 5HT staining. The descending system provides virtually the entire 5HT innervation of the dorsal horn, the intermediate zone, and the dorsal and lateral portions of the ventral horn. Fibers of the descending 5HT system course in the lateral funiculus, the dorsal portion of the ventral funiculus, and in the submeningeal zones of the dorsal and lateral aspects of the spinal cord. This projection primarily originates from the 5HT cell groups of the caudal rhombencephalon (groups II and III; Ritchie et al., '83), with a minor contribution from group IV in the rostral rhombencephalon. The organization of the descending 5HT system in stingrays is remarkably similar to that of mammals. The intrinsic spinal 5HT system consists of cells distributed in the ventromedial spinal cord that have processes extending longitudinally in a ventral submeningeal fiber network. Fibers were traced from the submeningeal system to the ventral horn, where varicose processes were restricted largely to the neuropil ventral to the somata of the fin motoneurons. The existence of a well-defined intrinsic 5HT system in stingrays supports the hypothesis that such a system exists in the spinal cords of a variety of vertebrates.

Central connections of ventral root afferents as demonstrated by the HRP method.

To study the central connections of ventral root afferents, horseradish peroxidase was injected into the lumbosacral spinal cord of the cat and the appropriate dorsal root ganglia were examined from segments with (1) both dorsal and ventral roots intact, (2) both roots sectioned, and (3) only the dorsal root sectioned. The key finding was that a number of labeled cells (up to 133) were observed in the ganglion after dorsal rhizotomy. We interpret these findings to mean that the central processes of the labeled cells projected to the spinal cord through the ventral root. As expected, when both roots were cut, almost no cells were labeled and when both roots were intact, there were large numbers of labeled cells. Of the labeled cells observed in ganglia after dorsal rhizotomy, all were found to be within the ganglion itself. No labeled aberrant dorsal root ganglion cells could be found.

Nuclei in which functionally identified spinothalamic tract neurons terminate.

The approximate level of termination of the axons of individual, functionally characterized spinothalamic tract neurons within the monkey thalmus was mapped by antidromic activation using a monopolar electrode which was moved in a systematic grid of tracks through the thalamus. The course of individual axons could be followed through several thalamic levels, and in a few cases branches to both the VPL nucleus and to the intralaminar nuclei were demonstrated. Most of the axons studied, however, projected just to the VPLc or VPLo nuclei. The spinothalamic tract cells that projected to the VPLc nucleus included representative of all known functional categories: low threshold, wide dynamic range, high threshold and "deep." It is speculated that these different classes of spinothalamic projections could make contributions to such sensory modalities as touch, proprioception and pain.

Funicular trajectories of brainstem neurons projecting to the lumbar spinal cord in the monkey (Macaca fascicularis): a retrograde labeling study.

Brainstem nuclei projecting to the lumbar spinal cord in the monkey were identified by using horseradish peroxidase and the fluorescent dye granular blue. These retrogradely transported tracers were used in fluid and/or gel forms to determine the funicular trajectories of the brainstem-spinal projections. The major descending components of the dorsal funiculus arose from the n. gracilis, n. cuneatus, and the n. of the solitary tract. Major components of the dorsolateral funiculus (DLF) came from the raphe complex, medullary and pontine reticular formation, locus coeruleus, Edinger-Westphal n., and red n. Other nuclei giving rise to minor contributions to the DLF included n. gracilis, n. cuneatus, n. of the solitary tract, medial and spinal vestibular n., subcoeruleus, periaqueductal gray, interstitial n. of Cajal, n. of Darkschewitsch, and the anteromedian n. The major components of ventral cord paths (ventrolateral and ventral funiculi) arose from the raphe complex, the medullary and pontine reticular formation, lateral and spinal vestibular n., and the coerulean complex. Minor contributions to the ventral paths descended from the dorsal motor n. of X, n. of the solitary tract, medial vestibular n., paralemniscal reticular formation, dorsal parabrachial n., n. cuneiformis, periaqueductal gray, Kölliker-Fuse n., and red n. The possible functional implications of the funicular distribution of these descending pathways are discussed from the perspective of descending inhibition and pain modulation.

The distribution of serotonin in the CNS of an elasmobranch fish: immunocytochemical and biochemical studies in the Atlantic stingray, Dasyatis sabina.

The distribution of serotonin (5HT) in the brain of the Atlantic stingray was studied with peroxidase-antiperoxidase immunocytochemistry and high-pressure liquid chromatography. The regional concentrations of 5HT determined for this stingray fell within the range of values previously reported for fishes. A consistent trend in vertebrates for the hypothalamus and midbrain to have the highest concentrations and the cerebellum the lowest was confirmed in stingrays. Neuronal cell bodies and processes exhibiting 5HT-like immunoreactivity were distributed in variable densities throughout the neuraxis. Ten groups of 5HT cells were described: (I) spinal cord, (II-IV) rhombencephalon, (V, VI) mesencephalon, (VII, VIII) prosencephalon, (IX) pituitary, and (X) retina. There were three noteworthy features of the 5HT system in the Atlantic stingray: (1) 5HT cells were demonstrated in virtually every location in which 5HT-containing cells have been described or alluded to in the previous literature. The demonstration of immunopositive cells in the spinal cord, the retina, and the pars distalis of the pituitary suggests that 5HT may be an intrinsic neurotransmitter (or hormone) in these regions. (2) The distribution of 5HT cells in the brainstem shared many similarities with that in other vertebrates. However, there were many 5HT cells outside of the raphe nuclei, in the lateral tegmentum. It appears that the hypothesis that "lateralization" of the 5HT system is an advanced evolutionary trend cannot be supported. (3) 5HT fibers and terminals were more widely distributed in the Atlantic stingray brain than has been reported for other nonmammalian vertebrates on the basis of histofluorescence. It appears that this feature of the 5HT system arose early in phylogeny, and that the use of immunohistochemistry might reveal a more general occurrence of widespread 5HT fibers and terminals.

Facilitation of the response of primate spinothalamic cells to cold and to tactile stimuli by noxious heating of the skin.

Spinothalamic tract cells in anesthetized monkeys were found to respond to noxious cold stimuli (18/19 cells tested), as well as to noxious heat and noxious mechanical stimuli. Responses to repetition of the noxious cold stimuli after a series of noxious heat stimuli were enhanced. However, subtraction of the enhanced background activity that resulted from damage of the skin revealed that the enhanced response to noxious cold stimuli were due to superposition of the original responses upon an enhanced background activity, rather than to sensitization of the responses to noxious cold stimuli per se. Furthermore, the responses to innocuous mechanical stimuli applied either within the area that was damaged or outside this area were enhanced, provided the noxious heat was applied for a long enough time. Thus, damage to a region of skin can result in enhanced responsiveness of spinothalamic cells to stimuli applied in an undamaged region of the receptive field. The possible relationship between these observations and cutaneous hyperalgesia is discussed.

The relationship of the medullary catecholamine containing neurones to the vagal motor nuclei.

We have re-examined in the rat the nuclear localization of the medullary catecholamine-containing cell groups (A1 and A2) and their relation to the vagal motor nuclei using a double labeling method. The vagal nuclei were defined by the retrograde transport of horseradish peroxidase applied to the cervical vagus, and noradrenergic and adrenergic neurons were stained with the peroxidase-antiperoxidase immunocytochemical method using an antibody to dopamine beta-hydrolase. The method allows visualization of both labels within single neurons. The neurons of the A2 group are primarily distributed in both the nucleus of the solitary tract and the dorsal motor nucleus of the vagus in a complex interrelationship that depends on the rostrocaudal level. Caudal to the obex, cells of the dorsal motor nucleus of the vagus are scattered among cells immunoreactive for dopamine beta-hydroxylase in the area considered to be the commissural subnucleus of the nucleus of the solitary tract. At levels near and slightly rostral to the obex, the dopamine beta-hydroxylase-positive cells are largely confined to nucleus of the solitary tract. However, the rostral third of the A2 group lies predominantly within dorsal motor nucleus, as defined by horseradish peroxidase labeled cells, with only a few cells in the nucleus of the solitary tract. A subset of the dopamine beta-hydroxylase positive cells within the rostral dorsal motor nucleus of the vagus are also vagal efferents. Our results suggest that a second population of dopamine beta-hydroxylase positive vagal efferents may exist ventrolaterally where neurons of the AI cell group intermingle with those of nucleus ambiguus.