Supraspinal modulation of nociception in awake rats by stimulation of the dorsal column nuclei

Rats with bilateral dorsal column (DC) lesions and chronic placement of stimulating electrodes on DC nuclei (DCN) and sham-operated rats were studied using the tail immersion test for phasic pain and the formalin test for tonic pain. DCN stimulation produced a clear inhibitory effect, through a supraspinal loop, in both phasic and tonic pain in awake rats.     

The morphology of dorsal column postsynaptic spinomedullary neurons in the cat

Dorsal column postsynaptic (DCPS) spinomedullary neurons from the cat's lumbosacral enlargement were identified by antidromic stimulation of the cervical dorsal columns and stained intracellularly with horseradish peroxidase. The cell bodies were located in laminae III-IV. Their dendritic arbors were elongated rostrocaudally but narrow mediolaterally. On the average, the arbors were ×5 longer than they were wide. Most of the neurons had nearly all of their dendrites in laminae III-IV and some of the neurons had, in addition, a considerable amount of dendritic surface area in lamina V. Only one neuron had more than a very small amount of dendritic surface area dorsal to lamina III. Seven of the neurons had unmyelinated axon collaterals that arborized extensively and issued varicosity-bearing terminal branches in laminae III-V, both within and beneath their dendritic territories. All of the neurons were excited by myelinated, low-threshold mechanoreceptors. Since the rostrocaudally elongated and mediolaterally narrow dendritic arbors of DCPS neurons are in register with the laminae III-IV terminal distributions of myelinated, low-threshold mechanoreceptors, it is probable that this excitation arises from a monosynaptic and topographically discrete innervation. About one-half of the DCPS neurons were also excited by noxious stimuli. It is probable that this excitation is accomplished by a polysynaptic distributions are largely or completely separate.     

Functional organization and connectivity of the dorsal column nuclei complex reveals a sensorimotor integration and distribution hub

The dorsal column nuclei complex (DCN‐complex) includes the dorsal column nuclei (DCN, referring to the gracile and cuneate nuclei collectively), external cuneate, X, and Z nuclei, and the median accessory nucleus. The DCN are organized by both somatotopy and modality, and have a diverse range of afferent inputs and projection targets. The functional organization and connectivity of the DCN implicate them in a variety of sensorimotor functions, beyond their commonly accepted role in processing and transmitting somatosensory information to the thalamus, yet this is largely underappreciated in the literature. To consolidate insights into their sensorimotor functions, this review examines the morphology, organization, and connectivity of the DCN and their associated nuclei. First, we briefly discuss the receptors, afferent fibers, and pathways involved in conveying tactile and proprioceptive information to the DCN. Next, we review the modality and somatotopic arrangements of the remaining constituents of the DCN‐complex. Finally, we examine and discuss the functional implications of the myriad of DCN‐complex projection targets throughout the diencephalon, midbrain, and hindbrain, in addition to their modulatory inputs from the cortex. The organization and connectivity of the DCN‐complex suggest that these nuclei should be considered a complex integration and distribution hub for sensorimotor information.     

Dorsal column stimulation in pain treatment

Thirty-five patients with chronic pain were treated by electrical stimulation of the dorsal columns through implanted epidural electrodes. In 5, such stimulation had minimal pain-reducing effect during an immediate postoperative trial period, and the electrodes were removed. In the remaining 30, an electrode system for chronic stimulation was implanted. Four of these died of related or unrelated causes. The electrode system was removed in 2 patients due to infections, and in 1 due to mechanical discomfort. The remaining 23 patients answered a questionnaire concerning the pain reducing effect of stimulation (4-60 months postoperatively, median 15 months). The group as a whole estimated the pain reducing effect of stimulation as weak. Only 10 patients (43.5%) used the stimulator regularly. Even in these patients, the pain-reducing effect was limited. Chronic back pain after repeated back surgery responded relatively better to stimulation than did the other cases. Phantom limb pain was most resistant. The modest results suggest future restriction on the use of such stimulation.     

Postsynaptic dorsal column pathway of the rat. III. Distribution of ascending afferent fibers

The distribution in the dorsal column nuclei (DCn) of post-synaptic dorsal column (PSDC) fibers was examined in rats following injections of Phaseolus vulgaris leucoagglutinin (PHA-L) in the spinal cord. Lemniscal neurons in the DCn were retrogradely labeled in the same animals by injecting the thalamus with Fluoro-Gold. In some experiments, primary afferent fibers were also labeled by injecting dorsal root ganglia with choleragenoid-conjugated HRP. Injections of PHA-L into the cervical enlargement labeled many fibers and varicosities throughout most of the ipsilateral cuneate nucleus. Labeled fibers were also present in the external cuneate and internal basilar nuclei. Injections of PHA-L into thoracic cord labeled fibers and varicosities in the medial cuneate and lateral gracile nuclei, as well as the external cuneate nucleus. Injections into the lumbar enlargement labeled fibers and varicosities throughout most of the gracile nucleus. Injections in sacral cord labeled fibers in the most medial part of the gracile nucleus. Dense labeling of PSDC fibers was found in areas with high densities of retrogradely labeled lemniscal neurons and areas with high densities of primary afferent fibers. In all regions of the DCn and in the external cuneate nucleus, fibers and varicosities labeled for PHA-L were seen in apposition to retrogradely labeled lemniscal cells. The distribution of postsynaptic afferent fibers in the DCn of the rat and its relationship to lemniscal neurons and primary afferent fibers contrast sharply with these features in cats.     

Comparison of the effects of dorsal rhizotomy or dorsal column transection on motor performance of monkeys

Macaca speciosa monkeys were trained to project either forelimb from variable start positions to various points on a frontal plane. The end points of these ballistic movements were visible before (1 to 5 s) but not during the trials; similarly, the responding limb was not visible to the monkeys during the timed trial period (from limb release to goal contact). Forelimb deafferentation by dorsal rhizotomy (C3 through T3) produced a severe deficit in the efficiency of limb projection; trial times were elevated by a factor of 7 or more. With daily practice, goal-directed movements of the affected limb began to recover (32 or 43 days after surgery) to a plateau that was maintained for 100 days or more. A late improvement period brought the monkeys to a performance level 2.7 times slower than control values. Subsequent lesion of the spinal dorsal columns (contralateral to the deafferentation) produced a forelimb deficit of lesser magnitude but similar time course. Analysis of these and other data suggest that the capacity for motoric recovery is quite substantial following drastic reduction of relevant sensory cues, but a large amount of task-specific practice is required.     

Supraspinal inhibition of trigeminal input into subnucleus caudalis by dorsal column stimulation

In decorticate-decerebellate cats with cervical spinal cuts which isolated the dorsal columns (DCs), conditioning volleys travelling in the DCs inhibited activities of trigemenal caudalis neurons. This inhibition was most likely mediated through a brainstem loop and was particularly effective on nociceptive-driven neurons.     

Changes in c-fos expression in the dorsal column-medial lemniscal system following dorsal column lesions

Transection of the hindlimb dorsal column fibers leads to a partial deafferentation of the neurons of the nucleus gracilis, the first relay of the ascending dorsal column-medial lemniscal (DC-ML) neural network. In response to the deafferentation, a synaptic renewal cycle is initiated and the neurons of the nucleus gracilis atrophy. The present study examines the molecular changes that occur in synaptic relays of the ascending DC-ML following a hindlimb dorsal column transection. Rats were sacrificed 0.5, 1, 24, or 72 hours postlesion. Steady-state levels of mRNA coding for c-fos were significantly elevated only at 24 hours postlesion in caudal dorsal medulla, which contains the nucleus gracilis. The increased c-fos is neuronal in origin since there is an increased level of c-fos immunoreactivity in both the cluster neurons and the interneurons of the nucleus gracilis. In the third relay of the DC-ML system, the somatomotor cortex, levels of c-fos mRNA were significantly decreased 72 hours postlesion. These data indicate that lesions of the hindlimb dorsal column fibers have transneuronal effects on gene expression that extend to at least the third synaptic relay in the DC-ML system. © 1995 Wiley-Liss, Inc.

1 This article is a US Government work and, as such, is in the public domain in the United States of America.

     

Mechanisms of sensitization of the response of single dorsal root ganglion cells from adult rat to noxious heat

We investigated the regulation by nerve growth factor of the response of sensory neurons to noxious heat (>43 degrees C). In dissociated dorsal root ganglion neurons (<30 micro m) from adult rat we demonstrated, using perforated patch clamp recording, that the inward current elicited in response to noxious heating is enhanced by nerve growth factor and reduced by capsazepine. The tachyphylaxis observed in response to the second of two heat pulses was reversed in most cells when nerve growth factor was introduced into the medium during the 5 min between the two heat stimuli, similar to findings using capsaicin [X. Shu & L.M. Mendell (1999) Neurosci. Lett.274, 159-162]. The threshold temperature did not change systematically after nerve growth factor. Using antibodies to TRPV1 and trkA in a subset of cells from which we recorded, we found a virtually perfect correlation between expression of TRPV1 and sensitivity to noxious heat. In addition, trkA expression was perfectly correlated with the ability of nerve growth factor to reverse tachyphylaxis. Thus, this physiological test is a reliable measure of trkA expression in cells sensitive to noxious heat. In agreement with studies in heterologous cells expressing trkA and TRPV1, pharmacologically blocking phospholipase C abolished the effect of nerve growth factor on heat-evoked currents in cells verified to express trkA. We conclude that the response of dorsal root ganglion neurons to noxious heat is conditioned by nerve growth factor in the same way as their response to capsaicin and that these responses require the presence of trkA and TRPV1.     

Dorsal column postsynaptic neurons in the cat are excited by myelinated nociceptors

Some (25-50%) dorsal column postsynaptic (DCPS) neurons respond only to innocuous mechanical stimuli; the remainder (50-75%) responds to both innocuous and noxious mechanical stimuli. Those that respond to noxious mechanical stimuli (pinch) are assumed to be excited by input from nociceptive primary afferents, but it is conceivable that their pinch-evoked responses are produced by the inadvertent activation of those low-threshold mechanoreceptive primary afferents that respond to stretching the skin. Because nociceptive primary afferents respond reliably to noxious heat and low-threshold mechanoreceptors do not, we tested DCPS neurons in the cat lumbar spinal cord with a series of noxious heat stimuli (48 degrees C or 50 degrees C-56 degrees C; 30 s duration). Seven of eight pinch-responsive neurons responded to noxious heat, but only after their receptive fields had been sensitized by prolonged or repeated heating. The results show that (1) many DCPS neurons in the cat are excited by nociceptive primary afferents and (2) these nociceptive afferents are probably myelinated high-threshold mechanoreceptors.     

Extra- and intracellular recordings from dorsal column postsynaptic spinomedullary neurons in the cat

Dorsal column postsynaptic (DCPS) spinomedullary neurons in the dorsal horn of spinal segments L6-S1 of adult cats anesthetized with sodium pentobarbital were identified by antidromic stimulation of cervical dorsal columns that were dissected free of, and electrically isolated from, the rest of the spinal cord. The neurons were categorized with respect to natural stimulation of their cutaneous receptive fields. An equal number of low-threshold mechanoreceptive and wide-dynamic-range neurons were found. No DCPS neurons could be classified as nociceptive-specific. All neurons received input from low-threshold mechanoreceptors with myelinated axons. There was no evidence that any neurons received monosynaptic input from unmyelinated, primary afferent fibers. The average conduction velocity of the antidromic responses was 45.7 m/s. Nearly half of the DCPS cells showed an antidromic spike followed by synaptically driven responses that were probably evoked by antidromic invasion into the intraspinal collaterals of A-beta primary afferent fibers that ascended the dorsal columns. Intracellularly recorded synaptic responses of DCPS neurons to dorsal column and receptive field stimulation usually consisted of an EPSP with overriding spike potentials followed by a prolonged IPSP whose amplitude decreased markedly as the stimulus frequency was increased in the range of 5 to 30 Hz. The results indicate that DCPS neurons constitute a projection system capable of signaling innocuous and tissue-damaging mechanical stimuli. The DCPS projection may play a role in the modulation of touch and pain perception.     

Activation of transient receptor potential vanilloid 2‐expressing primary afferents stimulates synaptic transmission in the deep dorsal horn of the rat spinal cord and elicits mechanical hyperalgesia

Probenecid, an agonist of transient receptor vanilloid (TRPV) type 2, was used to evaluate the effects of TRPV2 activation on excitatory and inhibitory synaptic transmission in the dorsal horn (DH) of the rat spinal cord and on nociceptive reflexes induced by thermal heat and mechanical stimuli. The effects of probenecid were compared with those of capsaicin, a TRPV1 agonist. Calcium imaging experiments on rat dorsal root ganglion (DRG) and DH cultures indicated that functional TRPV2 and TRPV1 were expressed by essentially non‐overlapping subpopulations of DRG neurons, but were absent from DH neurons and DH and DRG glial cells. Pretreatment of DRG cultures with small interfering RNAs against TRPV2 suppressed the responses to probenecid. Patch‐clamp recordings from spinal cord slices showed that probenecid and capsaicin increased the frequencies of spontaneous excitatory postsynaptic currents (sEPSCs) and spontaneous inhibitory postsynaptic currents in a subset of laminae III–V neurons. In contrast to capsaicin, probenecid failed to stimulate synaptic transmission in lamina II. Intrathecal or intraplantar injections of probenecid induced mechanical hyperalgesia/allodynia without affecting nociceptive heat responses. Capsaicin induced both mechanical hyperalgesia/allodynia and heat hyperalgesia. Activation of TRPV1 or TRPV2 in distinct sets of primary afferents increased the sEPSC frequencies in a largely common population of DH neurons in laminae III–V, and might underlie the development of mechanical hypersensitivity following probenecid or capsaicin treatment. However, only TRPV1‐expressing afferents facilitated excitatory and/or inhibitory transmission in a subpopulation of lamina II neurons, and this phenomenon might be correlated with the induction of thermal heat hyperalgesia.     

Inhibition of dorsal column by nuclei stimulation of trigeminal afferents in decerebrate—decerebellate cats

In decerebrate—decerebellate cats, stimulation of trigeminal afferents inhibited neurons in dorsal column (DC) nuclei driven by activation of DC input and produced primary afferent depolarization in DC primary afferent terminals. This inhibition was most likely mediated by a trigeminal—brainstem—DC nuclear loop.     

Patterns of afferentation in rat ventroposterolateral nucleus after thoracic dorsal column lesions

The secondary transneuronal effect of bilateral, dorsal column lesion (T12) on the frequency of boutons on cells in the lateral ventroposterolateral nucleus (VPL) was studied. Rats were killed 1, 2, 3, 7, 14, 30, 45, 60, 90, or 120 days postoperative (DPO), and bouton counts (Rasmussen stain) were made on the soma, or along 5- and 10-μm segments of the proximal dendrite branching from soma, in the rostral two-thirds of the VPL. The soma diameter also was measured of those neurons chosen for bouton counts on the circumference of the soma. Significant, increased afferentiation along a 5-μm segment of the proximal dendrite occurred during the first 3 days postlesion compared with longer survival times. Along the 10-μm segment, as at 5 μm on the proximal dendrite, bouton counts at 2 DPO were significantly higher than at 30 and 120 DPO. This rapid change in afferentation during the first week postlesion mirrors similar effects reported in the nucleus gracilis of these same cases. Bouton counts along the 5-μm segment of the proximal dendrite decreased significantly to less than normal at 30 and 120 DPO, indicating the nonstable and protracted character of afferentation after the distal cord lesion. A significant, positive correlation was obtained between boutons on the soma and proximal dendrite (5 μm segment) in cases with lesions, suggesting parallel innervation patterns for these adjacent neuronal components during 120 postoperative days. There was no significant change in the bouton counts or diameter of the soma during postoperative days. These results suggest that lesion of the thoracic dorsal column may result in subsequent rapid bouton patterning on the lateral VPL neurons, presumably mediated by their medial lemniscal afferent fibers. The postlesion source(s) of afferentation remains an intriguing, and as yet unanswered, question.     

Changes in dynamin and actin mRNA expression in the dorsal column-medial lemniscal system following dorsal column lesion

Transection of the third cervical hindlimb dorsal column nerve fibers in the spinal cord leads to a partial deafferentation atrophy of the neurons of the ascending dorsal column-medial lemniscal neural network (DC-ML) up to the cortex. We now examine the alteration of the steady-state level mRNA coding for the synaptic vesicle protein, dynamin I, and the cytoskeletal protein β-actin as early indicators of direct and trans-synaptic changes in the relay nuclei of the DC-ML. Rats were sacrificed at 6, 24, 72, and 240 hr after C3 hindlimb dorsal column or sham lesion. By 24 hr, there are changes in the steady-state levels of mRNA coding for both dynamin I and β-actin in regions of the brain containing the first (nucleus gracilis of medulla) and third synaptic relays (cortex). β-actin mRNA is increased at both 6 and 24 hr in the nucleus gracilis. The changes in dynamin I mRNA in the nucleus gracilis are early and biphasic, elevated at 6 hr but decreased compared to sham by 24 hr. In both regions, the initial fluctuations of dynamin I and β-actin mRNA levels are transient. By 72 hr, the levels are no different from those of sham-lesioned animals. In the somatomotor cortex, there is an additional increase in β-actin mRNA levels at 240 hr. The increased steady-state levels of dynamin and actin mRNA following a hindlimb dorsal column lesion suggest that increased synaptic vesicle recycling and actin cytoskeleton rearrangement are some of the early responses to deafferentation made by the neurons of the DC-ML synaptic relays. © 1996 Wiley-Liss, Inc.     

In vitro electrophysiological properties of rat dorsal column nuclei neurons.

The dorsal column nuclei include the gracile and cuneate nuclei, which receive somatosensory information from the periphery and project to the ventroposterior nucleus of the contralateral thalamus. The aim of this study was to determine the electrophysiological and morphological characteristics of the neurons of the dorsal column nuclei and to identify synaptic events evoked by electrical stimulation of the dorsal column, using an in vitro slice preparation. The results show two types of neurons, termed type I and II. A repolarizing sag distinguished type I cells during hyperpolarizing current injection, suggesting the activation of a Q-current. Moreover, type I cells, but not type II cells, were capable of maintaining spontaneous rhythmic activity at 9-15 Hz. Both types of cells displayed a delay in their return to the resting membrane potential following hyperpolarizing current pulses, indicating the existence of an A-current. Electrical stimuli applied to the dorsal column elicited brief EPSPs and IPSPs in both cell types. EPSPs were abolished by 6-cyano-7-nitroquinoxaline-2,3-dione, indicating that they were mediated through non-NMDA receptors. IPSPs were blocked by picrotoxin, implying the activation of GABAA receptors. Intracellular staining with carboxyfluoresceine revealed that type I neurons had elongated somas and primary dendrites that extended radially. Type II cells were smaller and had round somas with few primary dendrites, most of them emerging from one pole of the soma. The axon of many type I neurons was stained and could be followed running ventrally and in rostral direction.     

Spinal cord neuron inputs to the cuneate nucleus that partially survive dorsal column lesions: A pathway that could contribute to recovery after spinal cord injury

Dorsal column lesions at a high cervical level deprive the cuneate nucleus and much of the somatosensory system of its major cutaneous inputs. Over weeks of recovery, much of the hand representations in the contralateral cortex are reactivated. One possibility for such cortical reactivation by hand afferents is that preserved second‐order spinal cord neurons reach the cuneate nucleus through pathways that circumvent the dorsal column lesions, contributing to cortical reactivation in an increasingly effective manner over time. To evaluate this possibility, we first injected anatomical tracers into the cuneate nucleus and plotted the distributions of labeled spinal cord neurons and fibers in control monkeys. Large numbers of neurons in the dorsal horn of the cervical spinal cord were labeled, especially ipsilaterally in lamina IV. Labeled fibers were distributed in the cuneate fasciculus and lateral funiculus. In three other squirrel monkeys, unilateral dorsal column lesions were placed at the cervical segment 4 level and tracers were injected into the ipsilateral cuneate nucleus. Two weeks later, a largely unresponsive hand representation in contralateral somatosensory cortex confirmed the effectiveness of the dorsal column lesion. However, tracer injections in the cuneate nucleus labeled only about 5% of the normal number of dorsal horn neurons, mainly in lamina IV, below the level of lesions. Our results revealed a small second‐order pathway to the cuneate nucleus that survives high cervical dorsal column lesions by traveling in the lateral funiculus. This could be important for cortical reactivation by hand afferents, and recovery of hand use. J. Comp. Neurol. 523:2138–2160, 2015. © 2015 Wiley Periodicals, Inc.     

Clinical impact of intraoperative CCEP monitoring in evaluating the dorsal language white matter pathway

In order to preserve postoperative language function, we recently proposed a new intraoperative method to monitor the integrity of the dorsal language pathway (arcuate fasciculus; AF) using cortico–cortical evoked potentials (CCEPs). Based on further investigations (20 patients, 21 CCEP investigations), including patients who were not suitable for awake surgery (five CCEP investigations) or those without preoperative neuroimaging data (eight CCEP investigations including four with untraceable tractography due to brain edema), we attempted to clarify the clinical impact of this new intraoperative method. We monitored the integrity of AF by stimulating the anterior perisylvian language area (AL) by recording CCEPs from the posterior perisylvian language area (PL) consecutively during both general anesthesia and awake condition. After tumor resection, single‐pulse electrical stimuli were also applied to the floor of the removal cavity to record subcortico‐cortical evoked potentials (SCEPs) at AL and PL in 12 patients (12 SCEP investigations). We demonstrated that (1) intraoperative dorsal language network monitoring was feasible even when patients were not suitable for awake surgery or without preoperative neuroimaging studies, (2) CCEP is a dynamic marker of functional connectivity or integrity of AF, and CCEP N1 amplitude could even become larger after reduction of brain edema, (3) a 50% CCEP N1 amplitude decline might be a cut‐off value to prevent permanent language dysfunction due to impairment of AF, (4) a correspondence (<2.0 ms difference) of N1 onset latencies between CCEP and the sum of SCEPs indicates close proximity of the subcortical stimulus site to AF (<3.0 mm). Hum Brain Mapp 38:1977–1991, 2017 . © 2017 Wiley Periodicals, Inc.     

Enhancement of stimulation-induced ERK activation in the spinal dorsal horn and gracile nucleus neurons in rats with peripheral nerve injury

It has been suggested that low-threshold sensory pathways have an important role in the formation and maintenance of sensory abnormalities which are observed after peripheral nerve injury. In the present study, we examined the involvement of these pathways in the development of hyperexcitability after sciatic nerve injury (SNI) by detecting the intracellular signal molecule. The rats that received a transection of the sciatic nerve 7 days before were electrically stimulated at 0.1 mA and 3 mA in the proximal region of the nerve injury site. We found a small number of phosphorylated extracellular signal-regulated kinase (pERK)-labelled neurons in laminae I-II and III-IV of the spinal dorsal horn in the control rats after 0.1 mA stimulation. By contrast, there was a marked increased of pERK-labelled neurons both in the superficial laminae and laminae III-IV after the same stimulation in the SNI rats. Enhancement of ERK activation induced by 3 mA stimulation was also observed. Immunoreactivity of pERK in gracile nucleus neurons was also dramatically increased after 0.1 mA stimulation to the injured nerve. These data suggest that the rats with peripheral nerve injury had an increased responsiveness to the low- or high-threshold peripheral stimuli in I-II, III-IV and gracile nucleus neurons. Furthermore, SNI rats that received neonatal capsaicin treatment showed a decreased number of pERK neurons after 0.1 mA stimulation in the dorsal horn and gracile nucleus neurons compared to the control rats. Thus, C-fibres may contribute to the enhanced excitability of the low-threshold sensory neurons after peripheral nerve injury.