Mercury in hair of muskrats (Ondatra zibethicus) and mink (Mustela vison) from the U. S. Department of Energy Oak Ridge Reservation.
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Aging↗
Biomedical subjects
R T Stevens
Publications and source records attributed to R T Stevens.
Identification of functioning cortex using cortical optical imaging.
OBJECTIVE: The purpose of this study was to evaluate the technique of cortical optical imaging (COI) of intrinsic cortical optical signals related to neuronal activation. The specific goals of the study were to evaluate some of the technical aspects of COI and thus maximize the intensity of the image of this intrinsic signaling process and to determine the physiological reliability of COI in a well-defined animal system. METHODS: The intrinsic optical signal of activated whisker barrel cortex of rat was imaged using a computer-based technique for rapid acquisition of enhanced images. Single-unit microelectrode recordings of cortical neuronal responses to whisker movement were used to confirm the locations of the whisker barrels. RESULTS: Narrow band incident light at 600- to 610-nm wavelength was most effective for producing optical images. Images could be obtained during activation by a single long (40 s) stimulus or by averaging the signal generated by repeated shorter (1-8 s) stimuli. Focusing slightly below the cortical surface, minimizing movement, and abolishing extraneous light were all important in increasing the signal-to-noise ratio. The locations of whisker movement-evoked cortical activity determined using COI are consistent with the known functional anatomy of rat whisker barrel cortex. The images obtained with this experimental arrangement are shown to be accurate predictors of the location of neuronal activity determined by comparing the locations of active sites identified with COI with locations of areas of neuronal activity determined using single-cell recording techniques. CONCLUSIONS: COI is able to rapidly identify areas of cortex containing elicited neuronal activity. The technique allows cortical activation maps to be made rapidly with a very high degree of spatial resolution. COI is reliable and consistent over time. COI, if used carefully, holds promise as an intraoperative technique to study both human and experimental animal cortical function.
Animals↗
Direct spinal projections to limbic and striatal areas: anterograde transport studies from the upper cervical spinal cord and the cervical enlargement in squirrel monkey and rat.
With the anterograde tracers Phaseolus vulgaris-leucoagglutinin (PHA-L) and biotinylated dextranamine (BD), direct spinal connections from the upper cervical spinal cord (UC; C1 and C2) and the cervical enlargement (CE; C5-T1) were demonstrated in various striatal and limbic nuclei in both squirrel monkey and rat. Within each species and from each spinal level, the total number of terminals seen in the limbic and striatal areas was approximately 50-80% of the number seen within the thalamus. Labeled terminal structures were seen in the hypothalamic nuclei, ventral striatum, globus pallidus, amygdala, preoptic area, and septal nuclei. In both species, the number of labeled terminals in limbic and striatal regions was larger from UC than from CE, although the distributions to each nucleus varied with the specific lamina injected. In both species and from both UC and CE, approximately one-half of the projections to striatal and limbic areas terminated in the hypothalamus. The only region that demonstrated a topographical organization was the globus pallidus, where terminals from the CE were located dorsomedially to those from the UC. In the rat, UC and CE injections into the lateral dorsal horn and pericentral laminae resulted in the largest number of limbic and striatal terminations. The proportion of ipsilateral terminations was greatest when the medial laminae in the UC or the lateral dorsal horn in the CE received injections. Analysis of the morphology of these spinohypothalamic and spinotelencephalic terminals showed that, in the squirrel monkey, terminals from CE injections were larger than terminals from UC injections; no such size difference was evident in the rat. However, limbic and striatal terminals in the rat were generally larger than those in the squirrel monkey following injections into the UC or CE. The exact function of these direct spinal projections to various striatal and limbic areas in primates and in rodents remains to be determined. These findings, however, support recent imaging studies that suggest that the limbic system plays an important role in the mediation of chest pain, perhaps directly through these spinolimbic and spinostriatal pathways.
Animals↗
Spinothalamocortical projections to the secondary somatosensory cortex (SII) in squirrel monkey.
Anterograde labeling of the cervical spinothalamic tract was combined with retrograde labeling of thalamocortical cells projecting to the hand region of the second somatosensory cortex (hSII) to identify likely sites in the thalamus for processing and transmitting nociceptive information to hSII. Anterograde labeling of terminals was done with 2% WGA-HRP injections in the cervical enlargement; thalamocortical cells were retrogradely labeled with fluorescent tracers. In one experiment, the contralateral primary somatosensory cortex hand region (hSI) was injected to provide a direct comparison with hSII thalamic label. Both labeled cells and terminal-like structures were visualized in single thalamic sections and their numbers and positions quantitatively analyzed. The number of labeled cells within 100 microns from the STT terminals were counted as overlapping cells. Four thalamic nuclei, ventroposterior inferior (VPI), ventroposterior lateral (VPL), posterior nucleus (PO) and centrolateral nucleus (CL) combined to contain 86.5% of all hSII-projecting overlapping cells. Of all hSII-projecting thalamic overlapping cells, VPI contained the largest number (36.4% of the total) followed by the anterior portion of the posterior nuclear complex (POa; 20.4%), VPL (18.3%) and CL (11.4%). Results of the hSI injection show a different pattern of overlap in agreement with our earlier study. The relative distribution of overlapping cells was dependent on the antero-posterior position of the SII injections. The most anterior injections resulted in small numbers of labeled cells, with the majority of overlapping cells located in PO and CL. The more posterior injections resulted in overlapping cells mainly in VPI and VPL.(ABSTRACT TRUNCATED AT 250 WORDS)
Animals↗
Spinothalamocortical inputs nonpreferentially innervate the superficial and deep cortical layers of SI.
Using a combined anterograde and retrograde tracing technique, we examined the distribution pattern of the thalamocortical cells which projected to superficial layers of the hand region of the primary somatosensory cortex (hSI), and quantitatively analyzed the retrogradely labeled cells which putatively contacted terminals of the spinothalamic tract (STT) in the squirrel monkey and the macaque. Less than 25% of the superficial hSI projecting cells were putatively contacted by terminals of cervical enlargement spinothalamic neurons. These cells were primarily located in ventroposterior lateral, ventroposterior inferior and centrolateral nuclei. Although the number of superficial hSI projecting cells numbered less than 20% of the total hSI projecting cells, their patterns of location and their proportion of overlap with STT terminals within each thalamic nucleus were similar. It is suggested that the spinothalamic nociceptive information input to to the cortex equally accesses both superficial and deep SI.
Animals↗
The location of spinothalamic axons within spinal cord white matter in cat and squirrel monkey.
The locations of spinothalamic (STT) fibers in the spinal cord white matter have been identified in cat and squirrel monkey by light-microscopic visualization of labeled fibers following multiple thalamic injections of wheatgerm agglutinin conjugated to horseradish peroxidase. Thalamic injections were combined with either a constricting dural tie or an intraspinal injection of colchicine to facilitate axonal labeling at more rostral spinal levels. In the cat, the ventral-to-dorsal distribution of labeled STT fibers was bimodal. In the ventrolateral white matter, labeled axons were coarse in nature and were primarily concentrated peripherally. In the dorsolateral white matter, labeled STT axons consisted of fine-caliber fibers concentrated in the ventral portion of the dorsolateral funiculus and were equally distributed throughout the medial and lateral white matter. In the squirrel monkey, the distribution of STT fibers was unimodal, extending from the ventral surface of the spinal white matter to the ventralmost portion of the dorsolateral funiculus. As in the cat, however, the ventrally located axons were large and coarse and were primarily located in the peripheral white matter, whereas the dorsalmost STT fibers were of fine caliber and were distributed equally in the medial and lateral white matter.
Animals↗
A cryogenic device for reversibly blocking transmission through small regions of the spinal cord white matter.
A simple cryogenic device is described. This device is capable of cooling neural tissue in contact with the probe and maintaining the tissue at the desired temperature for extended periods of time. The cold probe can thereby reversibly block neural transmission through small portions of the spinal cord white matter. Interruption of axonal transmission is achieved by placing the tip of the device in contact with the exposed surface of the spinal cord and cooling the tip of the probe to -1 to +2 degrees C. The investigator monitors the tip temperature and adjusts the pump rate to maintain a constant tip temperature. The cross-sectional area under the probe where effective transmission block is achieved is about 1.5 mm2 which approximates the size of a single funiculus in the cat thoracic spinal cord. The cryogenic device was constructed for less than $700. The properties of this device were studied in physiologic experiments in cats. This device reversibly, selectively and repeatedly blocked the ascending mass action potential in the dorsolateral funiculus, transmission through ascending spinal axons in the dorsal columns, transmission through axons of spinal dorsal horn cells, the descending inhibitory input to the dorsal horn and the activity of thalamic nociceptive neurons. The reversible cold block effects on single units were observed for the duration of the experiments (up to 18 h) with no detectable damage to the underlying tissue. The physiologic effects of the cold block were usually reversed a few minutes after rewarming, although in some cases it took up to 40 min for the complete reversal of the cold block. This cryogenic device is useful for studying spinal cord pathways.
Animals↗
Medial, intralaminar, and lateral terminations of lumbar spinothalamic tract neurons: a fluorescent double-label study.
A dorsolateral spinothalamic tract (DSTT), consisting primarily of lamina I neurons, was confirmed in the cat lumbar spinal cord by the use of thalamic injections of fluorescent dyes combined with selective thoracic spinal cord lesions. In addition, collateralization of spinothalamic tract (STT) terminations to medial, lateral, and intralaminar thalamic regions was investigated by injections of two different fluorescent dyes into pairs of these regions. The results of this study indicate that less than 15% of cat lumbar STT neurons collateralize to more than one of the thalamic regions evaluated. Lumbar lamina I cells project to the lateral and to the medial thalamus (13% collateralize to these two regions) and have only a scant projection to the intralaminar thalamus. Lumbar laminae IV-VI STT cells are very few in cat and demonstrate almost no collateralization to multiple thalamic areas. Neurons of laminae VII-X project equally to the three thalamic regions evaluated, and approximately 10-14% of cells from this laminar group collateralize to any two of the thalamic sites evaluated.
Animals↗
The spinothalamic tract: an examination of the cells of origin of the dorsolateral and ventral spinothalamic pathways in cats.
The locations of spinothalamic neurons and the funicular trajectories of their axons were studied in cats by retrograde transport of horseradish peroxidase (HRP). Five animals were used as controls to determine the cervical and lumbar laminar distributions of neurons contributing to the spinothalamic tract. An additional eight animals were used to determine the funicular trajectories of the spinothalamic axons of lumbar neurons by utilizing a series of thoracic spinal cord lesions in conjunction with retrograde transport of HRP from the sensory thalamus. Three of these animals underwent midthoracic ventral quadrant lesions, four animals underwent midthoracic dorsolateral funiculus lesions, and one animal underwent total spinal cord transection sparing the dorsal columns. The locations of the cells containing the HRP reaction product were then determined after a 3- to 5-day survival time, and the patterns of labeled cell locations of the lesion groups were compared to the control group patterns. In the lesioned animals, the cervical spinothalamic cell locations were used as a control to confirm the uniformity of the injection sites, transport and tissue processing. The major finding of this study is that there exist two distinct components of the spinothalamic tract. The dorsolateral spinothalamic tract (DSTT) is made up of axons originating in contralateral spinal cord lamina I and has negligible contribution from the deeper spinal cord laminae. The axons of lamina I cells cross segmentally and ascend exclusively in the dorsolateral funiculus (DLF). The DSTT comprises approximately 25% of the total spinothalamic input from the lumbar enlargement. The ventral spinothalamic tract (VSTT) is made up of axons originating in spinal cord laminae IV-V and VII-X. Very few lamina I cells contribute axons to the VSTT. This crossed pathway ascends in the ventrolateral and ventromedial portions of the spinal cord. No cells contributing to the spinothalamic tract were identified in spinal cord segments caudal to a dorsal column sparing lesion, indicating that there are no spinothalamic tract axons traveling in the dorsal columns. These results expand the classical concept of information processing by the spinothalamic tract. The DSTT is made up of lamina I cell axons. All lamina I spinothalamic cells respond exclusively to noxious peripheral stimuli. Hence the DSTT is a major nociceptive-specific ascending spinal pathway, yet lies outside the confines normally assigned to the spinothalamic tract.(ABSTRACT TRUNCATED AT 400 WORDS)
Animals↗
Inhibition of dorsal-horn cell responses by stimulation of the Kölliker-Fuse nucleus.
The Kölliker-Fuse nucleus (KF) in the dorsolateral pons has been shown to be the major source of catecholamine innervation of the spinal cord. This has important implications in terms of pain control mechanisms, since catecholamine-mediated mechanisms are essential for the expression of opiate and other varieties of antinociception. This study examines the effects of KF stimulation on responses of dorsal-horn cells to innocuous and noxious cutaneous stimuli in anesthetized cats. Stimulation of the KF potently inhibits the responses of dorsal-horn cells to both noxious and innocuous stimuli. The threshold for the inhibitory effect is significantly lower for responses to noxious stimuli as opposed to innocuous stimuli. The inhibitory effect is specific to the stimulus site, as evidenced by a marked decrease in the effect following small changes in the position of the stimulating electrode in the brain stem. The latency of the effects indicates a bulbospinal conduction velocity of 4 to 5 m/sec, which is much slower than usual reticulospinal effects and is consistent with a catecholamine-mediated system. The dependence of KF-spinal inhibition on intact biogenic amines was tested by depleting the animals of these amines with reserpine pretreatment. Depletion of biogenic amines resulted in a significant decrease in the KF spinal inhibitory effects, suggesting their dependence on intact noradrenergic stores. The results of these studies are consistent with the idea that the KF-spinal system plays an important noradrenergic-dependent role in the brain-stem modulation of spinal processing of noxious, potentially painful stimuli.
Animals↗
Funicular course of catecholamine fibers innervating the lumbar spinal cord of the cat.
The purpose of this study was to determine the funicular location of descending catecholamine (CA) fibers innervating the lumbar spinal cord from the dorsolateral pons (DLP). The locations of catecholamine-containing cell bodies which project to the lumbar spinal cord were determined by combining the use of the retrogradely transported fluorescent dye, Evans Blue (EB), with the glyoxylic acid histofluorescence technique. Lumbar injections of Evans Blue were combined with thoracic lesions of the dorsolateral funiculi (DLF) or ventrolateral funiculi (VLF) in order to retrogradely label those CA-containing or non CA-containing cell bodies whose axons descend within the spared hemispinal cord. By this technique it was determined that descending CA fibers innervating the lumbar spinal cord of the cat project through both the DLF and the VLF. The nucleus subcoeruleus, the Kolliker-Fuse nucleus and the CA cell bodies in the area of A5 each contain a significant number of CA-containing cells whose fibers descend both within the DLF and the VLF, while the nucleus locus coeruleus projects to the lumbar cord primarily through the VLF. Catecholamine cells of the DLP innervate the lumbar spinal cord bilaterally, although there is an ipsilateral predominance. The CA-containing cells of the DLP which innervate the contralateral spinal cord were shown by ipsilateral or contralateral thoracic hemisection to decussate both above and below the thoracic lesion. Non-CA-containing cells from the DLP also crossed at all levels of the spinal cord; however, cells from the caudal pons had a larger number of cells which crossed above the thoracic lesion while cells of the more rostral pons had a larger number of cells which crossed below the lesion.
Animals↗
A dorsolateral spinothalamic pathway in cat.
A spinothalamic tract that courses in the dorsolateral funiculus of the spinal cord and originates almost exclusively from spinal lamina I neurons has been demonstrated in the cat by retrograde transport of horseradish peroxidase. This tract is of special interest because the course of this predominantly lamina I, contralateral projection lies outside the classical course of the spinothalamic tract and because most lamina I cells contributing to the spinothalamic tract have been shown by other investigators to respond exclusively to somatic noxious stimuli. This newly described tract has important implications in the processing of noxious stimuli.
Animals↗
Funicular location of ascending axons of lamina I cells in the cat spinal cord.
The laminar distribution of spinal cord neurons projecting suprasegmentally through different funiculi was determined in the cat using horseradish peroxidase (HRP) injections combined with selective spinal cord lesions. The lesions were designed to limit the caudal transport of HRP to either the ventral funiculi or the dorsolateral funiculus. HRP injections in the ventromedial or ventrolateral funiculi resulted in labeling primarily within laminae IV-VIII and a virtual lack of labeling within lamina I. When the dorsolateral funiculus was injected, 20-25% of all labeled cells were located in lamina I, bilaterally. These results demonstrate that the ascending lamina I projections are through the dorsolateral funiculus.
Afferent Pathways↗
Sources of the catecholaminergic innervation of the trigeminal nucleus caudalis in cat.
Injections of the retrogradely transported fluorescent dye, Evans blue, into the trigeminal nucleus caudalis were combined with the glyoxylic acid histofluorescence technique to determine the sources of catecholamine-containing varicosities innervating nucleus caudalis. Results indicate that the sources of this catecholamine innervation are widespread, originating from cell bodies throughout the brain stem including the medullary catecholamine cell groups as well as the noradrenergic nuclei of the dorsolateral pons, including locus ceruleus, subceruleus, Kölliker-Fuse, and the parabrachial nuclei. A small projection from the presumably dopaminergic neurons of the hypothalamus was also noted. The catecholamine innervation of n. caudalis in the cat is from widespread brain stem sources, a pattern different from the catecholamine innervation of the spinal cord, which receives its major catecholamine input from the Kölliker-Fuse nucleus.
Animals↗
Lumbar dorsal root potentials elicited by stimulation of nucleus locus coeruleus.
Lumbar dorsal root potentials (DRP) were elicited by nucleus locus coeruleus (LC) stimulation in the cat. Inhibition, by LC stimulation, of dorsal horn cells responding to noxious inputs corresponded in time with the DRPs evoked by LC stimulation. Comparing cutaneous stimulation-evoked DRPs with LC stimulation-evoked DRPs and their respective effects on dorsal horn single-unit activity suggested a shared segmental underlying mechanism and the possible involvement of the coeruleospinal system with that of a diffuse noxious inhibitory suprasegmental loop.
Animals↗
Catecholamine varicosities in cat dorsal root ganglion and spinal ventral roots.
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Adrenergic Fibers↗
Dorsolateral pontine inhibition of dorsal horn cell responses to cutaneous stimulation: lack of dependence on catecholaminergic systems in cat.
The effect of stimulating the dorsolateral pons (DLP) in the region of locus ceruleus (LC) on lumbar dorsal horn cell responses to innocuous and noxious cutaneous stimuli was assessed and the dependence of these effects on intact pontospinal catecholaminergic systems was tested in chloralose-anesthetized cats. DLP stimulation inhibited the responses of dorsal horn cells to both noxious and innocuous skin stimuli. The inhibitory effect was most prominent when the responses to noxious stimuli were tested. The thresholds for eliciting DLP-spinal inhibition were lowest (less than 30 microA) in the region of LC. The inhibitory effect was found in both ipsilateral and contralateral dorsal horns. The DLP-spinal inhibition was unaltered by depletion of spinal catecholamines brought about by repeated lumbar intrathecal administration of 6-hydroxydopamine or systemic administration of reserpine. We conclude that the DLP-dorsal horn inhibition is not related to a catecholaminergic ceruleospinal system in the cat and that the dependence of pain modulation by catecholamine systems is a reflection of other descending pathways.
Animals↗
Kölliker-Fuse nucleus: the principal source of pontine catecholaminergic cells projecting to the lumbar spinal cord of cat.
Using retrograde transport of the fluorescent dye Evans Blue (EB), in combination with glyoxylic acid histofluorescence, the ponto-spinal catecholaminergic pathways were investigated. The cells which contain catecholamine and project to the lumbar spinal cord of the cat are most densely concentrated in the Kölliker-Fuse nucleus. Locus coeruleus, the subcoeruleus area, and the parabrachial nuclei were found to have relatively few cells that both contain catecholamine and project to the lumbar spinal cord.
Animals↗