Primary afferent receptive field properties and neurotransmitter candidates in a vertebrate lacking unmyelinated fibers.
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Biomedical subjects
R B Leonard
Publications and source records attributed to R B Leonard.
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.
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Spinal cord pathways involved in initiation of swimming in the stingray, Dasyatis sabina: spinal cord stimulation and lesions.
In spinally transected stingrays, electrical stimulation of a site just ventral to the dorsal root entry zone or a site in the intermediate portions of the lateral funiculus produced rhythmic swimming like movements of the contralateral pectoral fin. Electromyographic (EMG) records collected during cord-stimulated rhythms had the same pattern of activity and sometimes the same intersegmental coordination as those collected during spontaneous swimming of the same animal. In paralyzed, high-spinal stingrays, the only stimulation sites that produced rhythmic activity (fictive swimming) in the pectoral fin motor nerves were in the intermediate portion of the lateral funiculus. The evoked rhythm occurred in the motor nerves that were contralateral to the stimulated side of the spinal cord. The effects of subtotal lesions of the rostral spinal cord on spontaneous swimming behavior were assessed by analysis of EMG records taken before and after the lesions were made. Severe deficits in swimming occurred after bilateral ablation of intermediate portions of the lateral funiculi. In agreement with previous results, the stimulation experiments indicate that the stingray spinal cord contains an inherent capacity to generate properly coordinated rhythmic swimming. The current experiments also suggest that the descending pathways(s) that normally functions to initiate swimming projects through the intermediate aspects of the lateral funiculi.
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Intracellular recording from pectoral fin motoneurons of the stingray Dasyatis sabina, an elasmobranch fish.
Intracellular recordings were made from antidromically identified pectoral fin motoneurons in unanesthetized, decerebrate stingrays (Dasyatis sabina). These recordings had the three all-or-none components seen in other vertebrate motoneuron recordings. About 25% of the impalements had resting membrane potentials that were greater than -80 mV, which is larger than those of motoneurons from other vertebrate species. A novel depolarizing afterpotential (DAP) is associated with the isolated action potential occurring at the first node of Ranvier of the axon (M-spike). Occlusion experiments exclude recurrent events as the source of this potential. A capacitive source for the DAP is postulated. Using morphological and passive electrical data on motoneurons from previous studies, calculations of the passive decay of the nodal spike indicate that the membrane resistance of the initial segment is low and nearly equal to that of nodal membrane. The soma-dendritic (SD) spike is followed by a prominent, humped delayed depolarization (DD). The DD is temporally associated with the onset of the action potential produced by the initial segment (IS spike). Sources of the long-lasting period of repolarization recorded with the IS spike, which may underlie the DD, are postulated. The afterhyperpolarization (AHP) of stingray motoneurons tends to be shorter and smaller in amplitude than that of other vertebrate motoneurons. A negligible conductance change was often found during the period following an SD spike. No significant correlation was found between AHP duration and axonal conduction velocity. The input conductance of stingray motoneurons ranged between 1.5 X 10(-7) and 13.3 X 10(-7) S. The relationship between input conductance and axonal conduction velocity was determined from 42 motoneurons. These data were fitted by a power function with an exponent of 1.7, indicating that, in terms of membrane conductance properties, large stingray motoneurons are simply scaled-up versions of the small motoneurons.
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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.
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Organization of spinal motor nuclei in the stingray, Dasyatis sabina.
The Atlantic stingray, Dasyatis sabina, has enlarged pectoral fins consisting of a series of antagonist dorsal (elevator) and ventral (depressor) muscles. Each muscle is divided into superficial and deep components. The retrograde transport of horseradish peroxidase (HRP) was used to determine the organization of motoneuron pools innervating fin and epaxial muscles. HRP applied to a single peripheral nerve labeled motoneurons within a single spinal segment. Following intramuscular injection of HRP, 3 distinct cell groups were identified in the transverse plane. Motoneurons innervating elevator muscles were lateral in the ventral horn, while motoneurons innervating depressor muscles were dorsomedial. The epaxial muscles were found to be innervated by a distinct cell column along the ventral border of the ventral horn. Separate injections of the superficial and deep bundles of the elevator muscle resulted in considerable overlap in the distribution of labeled motoneurons. Soma areas for both elevator and depressor motoneurons were unimodally distributed. The mean cell diameters were 33.6 and 31.8 micron respectively. Motoneurons innervating the superficial and deep bundles of elevator muscle also had similar size distributions. The location of motoneurons innervating elevator and depressor fin muscles in the stingray supports the hypothesis that motoneurons innervating muscle derived from the dorsal premuscle mass are located laterally in the ventral horn while motoneurons innervating muscle derived from the ventral premuscle mass are located medially.
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Immunocytochemical demonstration of serotonergic neurons and processes in the retina and optic nerve of the stingray, Dasyatis sabina.
Neurons and processes in the stingray retina can be stained using PAP immunohistochemistry and an antibody to serotonin, without pharmacological pretreatment. Most of the cell bodies are in the inner nuclear layer while the processes ramify in the inner plexiform layer suggesting the presence of a population of serotonin containing amacrine cells in this species. Scattered immunopositive axons were observed in the optic nerve from the optic chiasm to the optic nerve head.
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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.
Afferent Pathways↗
Swimming pattern in intact and decerebrated stingrays.
Stingrays swim with a rostral-to-caudal wave of elevation and depression of the enlarged pectoral fins. The present study compares the timing relations of swimming before and after decerebration in the same animals using electromyograms (EMGs). EMGs were recorded from several freely swimming, intact animals. Most recordings, however, were from stingrays suspended by vertebral clamps. Activity recorded from restrained animals exhibited the same timing relations as freely swimming animals. Therefore, the restrained preparation was used to compare locomotor rhythm in intact and decerebrate stingrays. The delay from the onset of activity recorded at a rostral segment to the onset of activity at successively caudal recording sites (intersegmental delay) was linearly related to the extent of the segmental separation. This relation was not changed by decerebration. Both the intersegmental delay and the EMG burst duration were linearly related to the period of the swim cycle. The intercepts for both relations fell around the origin. Therefore, stingray swimming demonstrates constant phase coupling. Neither the slopes nor the intercepts of the coupling relations were altered by decerebration. Histological examination of the fins indicates that at least two types of muscles exist. Superficial muscle is activated during slow swimming, while the deeper muscle is recruited during faster swimming.
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Swimming rhythm in decerebrated, paralyzed stingrays: normal and abnormal coupling.
Rhythmic motoneuronal activity was recorded from decerebrated, paralyzed stingrays and compared with electromyograms recorded from the same animals. Before and after paralysis, a rostral-to-caudal sequence of alternation occurred between dorsal (elevator) and ventral (depressor) efferents. The swimming pattern was thus observed in the absence of phasic afferent input, and this constitutes fictive locomotion. After paralysis, both the intersegmental delay (time between activation at progressively caudal recording sites) and the burst duration remained linearly related to the swim cycle period. In many instances, neither the slope nor the intercept was significantly altered by immobilization. The intercepts all fell near the origin, indicating that the fictive rhythm remains constant phase coupled. Although the swimming rhythm was obtained after paralysis, some differences occurred. These included fewer and shorter spontaneous sequences, a greater range of cycle periods, and longer burst durations. During fictive swimming, the burst duration:cycle period ratio usually increased to 0.53 from 0.39 observed before paralysis. Therefore, the silent periods seen between burst discharges in antagonist efferents during movement were often absent after paralysis. Mechanical stimulation of the tail reduced both cycle periods and burst durations; however, the burst:cycle ratio remained greater than or equal to 0.50. The linear relation between burst duration and cycle period found for spontaneous sequences was not changed by stimulation of the tail. During fictive swimming the inter- and intrasegmental coupling that characterizes stingray swimming becomes labile. Abnormal coupling appears more often during sequences with long swim cycles. Intrasegmental coupling is tighter than intersegmental coupling at any cycle period. Rhythmic activity at one segmental level can be independent of activity at other levels. This suggests that multiple oscillator circuits exist that are not dependent on propriospinal circuits interconnecting different segments. Rhythmicity in elevator and depressor motoneurons is not dependent on reciprocal connections between the circuitry driving the motor nuclei. Therefore, separate oscillators for elevators and depressors appear to be present within one spinal segment.
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Immunocytochemical demonstration of serotonergic cells, terminals and axons in the spinal cord of the stingray, Dasyatis sabina.
Serotonin-like immunoreactivity, as demonstrated by the PAP method, was contained within cells as well as fibers and terminals, in the spinal cord of the stingray. Serotonergic spinal neurons were seen on 43% of the sections examined and were restricted to the ventral white matter. Immunoreactive terminals and varicosities were densely distributed over the spinal gray matter at all segmental levels, and stained fibers were seen in all portions of the white matter with the exception of the medial anterior and dorsal funiculi.
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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.
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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.
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Induction of swimming in the high spinal stingray by L-DOPA.
Stingrays with high spinal transections, which do not spontaneously locomote, can be induced to swim by intravenous injection of L-DOPA. The L-DOPA-induced swim of the spinal animal is associated with patterns of EMG activity that appear similar to those of the spontaneous swim of the decerebrate preparation. However, in contrast to the decerebrate condition, the L-DOPA-induced cycles of swimming are slower and less vigorous. Furthermore, secondary periodicites and altered intersegmental timing relationships are also evident.
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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.
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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.
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Emergency department radiation accident protocol.
Every emergency department faces the potential problem of handling one or more victims of a radiation accident. While emergency departments near nuclear power plants or isotope production laboratories probably have a detailed protocol for such emergencies, a similar protocol is needed for the emergency department that may have to handle an isolated event, such as a vehicular accident that spills radioactive material and contaminates passengers or bystanders. This communication attempts to answer that need, presenting a step-by-step protocol for decontamination of a radiation victim, the rationale on which each step is based, a list of needed supplies, and a short summary of decorporation procedures that should be started in the emergency department.
Accidents, Occupational↗
Responses of neurons in primate ventral posterior lateral nucleus to noxious stimuli.
1. Recordings were made from the caudal part of the ventral posterior lateral (VPLc) nucleus of the thalamus in anesthetized macaque monkeys. In additon to many neurons that responded only to weak mechanical stimuli, scattered neurons were found that responded to both innocuous and noxious stimulation or just to noxious stimulation of the skin. A total of 73 such neurons were examined in 26 animals. 2. Noxious stimuli included strong mechanical stimuli (pressure, pinch, and squeezing with forceps) and graded noxious heat (from 35 degrees C adapting temperature to 43, 45, 47, and 50 degrees C). The responses of the VPLc neurons increased progressively with greater intensities of noxious stimulation. The stimulus-response function when noxious heat stimuli were used was a power function with an exponent greater than one. 3. Repetition of the noxious heat stimuli revealed sensitization of the responses of the thalamic neurons to such stimuli. The threshold for a response to noxious heat was lowered, and the responses to supra-threshold noxious heat stimuli were enhanced. 4. The responses of VPLc neurons to noxious heat stimuli adapted after reaching a peak discharge frequency. The rate of adaptation was slower for a stimulus of 50 degrees C than for one of 47 degrees C. 5. For the six neurons tested, responses to noxious heat were dependent on pathways ascending in the ventral part of the lateral funiculus contralateral to the receptive field (ipsilateral to the thalamic neuron). In two cases, the input to the thalamic neurons from axons of the dorsal column was also conveyed by way of a crossed pathway in the opposite ventral quadrant. In another case, access to the thalamic neuron by way of ascending dorsal column fibers was demonstrated. 6. The thalamic neurons had restricted contralateral receptive fields that were somatotopically organized. Neurons with receptive fields on the hindlimb were in the lateral part of the VPLc nucleus, whereas neurons with receptive fields on the forelimb were in medial VPLc. 7. Ninety percent of the VPLc neurons tested that responded to noxious stimuli could be activated antidromically by stimulation of the surface of SI sensory cortex. It was possible to confirm that many of these cells project to the SI sensory cortex by using microstimulation. Successful microstimulation points were either within the SI cortex or in the white matter just beneath the cortex. 8. We conclude that some neurons in the VPLc nucleus are capable of signaling noiceptive stimuli. The nociceptive information appears to reach these cells through the ventral part of the lateral funiculus on the side contralateral to the receptive field, presumably by way of the spinothalamic tract. The VPLc cells are somatotopically organized, and they are thalamocortical neurons that project to the VPLc nucleus and SI cortex play a role in nociception.
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