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Activation of Aβ-fibers is fundamental to numerous analgesic therapies, yet its effects on dorsal horn neuronal activity remain unclear. We used multiphoton microscopy of the genetically encoded calcium indicator GCaMP6s to characterize the effects of Aβ-fiber electrical stimulation (Aβ-ES) on neural activity. Specifically, we quantified somatic responses evoked by C-fiber intensity stimulation before and after a 10-minute train of dorsal root Aβ-ES in superficial dorsal horn (SDH) neurons, in mouse lumbar spinal cord. Aβ-ES did not alter C-fiber evoked activity when GCaMP6s was virally expressed in all neurons, in an intact lumbar spinal cord preparation. However, when we restricted the expression of GCaMP6s to excitatory or inhibitory populations, we observed that Aβ-ES modestly potentiated evoked activity of excitatory neurons and depressed that of inhibitory neurons. Aβ-ES had no significant effects in a slice preparation in either SDH population. A larger proportion of SDH neurons were activated by Aβ-ES when delivered at a root rostral or caudal to the segment where the imaging and C-fiber intensity stimulation occurred. Aβ-ES effects on excitatory and inhibitory populations depended on the root used. Our findings suggest that Aβ-ES differentially modulates lumbar spinal cord SDH populations in a cell type-, and input-specific manner. Furthermore, they underscore the importance of the Aβ-ES delivery site, suggesting that Aβ stimulation at a segment adjacent to where the pain is may improve analgesic efficacy.

Although nonsteroidal anti-inflammatory drugs are the first line of therapeutics for the treatment of mild to moderate somatic pain, they are not generally considered to be effective for neuropathic pain. In the current study, direct activation of spinal Toll-like 4 receptors (TLR4) by the intrathecal (IT) administration of KDO2 lipid A (KLA), the active component of lipopolysaccharide, elicits a robust tactile allodynia that is unresponsive to cyclooxygenase inhibition, despite elevated expression of cyclooxygenase metabolites in the spinal cord. Intrathecal KLA increases 12-lipoxygenase-mediated hepoxilin production in the lumbar spinal cord, concurrent with expression of the tactile allodynia. The TLR4-induced hepoxilin production was also observed in primary spinal microglia, but not in astrocytes, and was accompanied by increased microglial expression of the 12/15-lipoxygenase enzyme 15-LOX-1. Intrathecal KLA-induced tactile allodynia was completely prevented by spinal pretreatment with the 12/15-lipoxygenase inhibitor CDC or a selective antibody targeting rat 15-LOX-1. Similarly, pretreatment with the selective inhibitors ML127 or ML351 both reduced activity of the rat homolog of 15-LOX-1 heterologously expressed in HEK-293T cells and completely abrogated nonsteroidal anti-inflammatory drug-unresponsive allodynia in vivo after IT KLA. Finally, spinal 12/15-lipoxygenase inhibition by nordihydroguaiaretic acid (NDGA) both prevents phase II formalin flinching and reverses formalin-induced persistent tactile allodynia. Taken together, these findings suggest that spinal TLR4-mediated hyperpathic states are mediated at least in part through activation of microglial 15-LOX-1.

Metastatic bone pain is the single most common form of cancer pain and persists as a result of peripheral and central inflammatory, as well as neuropathic mechanisms. Here, we provide the first characterization of sphingolipid metabolism alterations in the spinal cord occurring during cancer-induced bone pain (CIBP). Following femoral arthrotomy and syngenic tumor implantation in mice, ceramides decreased with corresponding increases in sphingosine and the bioactive sphingolipid metabolite, sphingosine 1-phosphate (S1P). Intriguingly, de novo sphingolipid biosynthesis was increased as shown by the elevations of dihydro-ceramides and dihydro-S1P. We next identified the S1P receptor subtype 1 (S1PR1) as a novel target for therapeutic intervention. Intrathecal or systemic administration of the competitive and functional S1PR1 antagonists, TASP0277308 and FTY720/Fingolimod, respectively, attenuated cancer-induced spontaneous flinching and guarding. Inhibiting CIBP by systemic delivery of FTY720 did not result in antinociceptive tolerance over 7 days. FTY720 administration enhanced IL-10 in the lumbar ipsilateral spinal cord of CIBP animals and intrathecal injection of an IL-10 neutralizing antibody mitigated the ability of systemic FTY720 to reverse CIBP. FTY720 treatment was not associated with alterations in bone metabolism in vivo. Studies here identify a novel mechanism to inhibit bone cancer pain by blocking the actions of the bioactive metabolites S1P and dihydro-S1P in lumbar spinal cord induced by bone cancer and support potential fast-track clinical application of the FDA-approved drug, FTY720, as a therapeutic avenue for CIBP.

Background Neuronal death may be an overlooked and unaddressed component of disability following neonatal nerve injuries, such as obstetric brachial plexus injury. N-acetylcysteine and acetyl-L-carnitine improve survival of neurons after adult nerve injury, but it is unknown whether they improve survival after neonatal injury, when neurons are most susceptible to retrograde neuronal death. The authors' objective was to examine whether N-acetylcysteine or acetyl-L-carnitine treatment improves survival of neonatal motor or sensory neurons in a rat model of neonatal nerve injury. Methods Rat pups received either a sciatic nerve crush or transection injury at postnatal day 3 and were then randomized to receive either intraperitoneal vehicle (5% dextrose), N-acetylcysteine (750 mg/kg), or acetyl-L-carnitine (300 mg/kg) once or twice daily. Four weeks after injury, surviving neurons were retrograde-labeled with 4% Fluoro-Gold. The lumbar spinal cord and L4/L5 dorsal root ganglia were then harvested and sectioned to count surviving motor and sensory neurons. Results Transection and crush injuries resulted in significant motor and sensory neuron loss, with transection injury resulting in significantly less neuron survival. High-dose N-acetylcysteine (750 mg/kg twice daily) significantly increased motor neuron survival after neonatal sciatic nerve crush and transection injury. Neither N-acetylcysteine nor acetyl-L-carnitine treatment improved sensory neuron survival. Conclusions Proximal neonatal nerve injuries, such as obstetric brachial plexus injury, produce significant retrograde neuronal death after injury. High-dose N-acetylcysteine significantly increases motor neuron survival, which may improve functional outcomes after obstetrical brachial plexus injury.

Background Capping techniques have been used as a treatment modality for the prevention of neuroma formation and the management of neuropathic pain. However, the results are inconsistent and unpredictable. We hypothesize that this situation may be attributable, in part, to the disparities in the type of materials used to manufacturing of the conduits. Methods In this study, a rat model was used and the sciatic nerve was selected for evaluation. In 1 capping group, a sciatic nerve stump was capped with a nonaligned nanofiber conduit (the nonaligned group), whereas in a second capping group, the conduit was made of aligned nanofibers (the aligned group). In another group, the sciatic nerve stump was not capped as a control (the control group). The results of autotomy behavior, extent of neuroma formation, histological changes in the neuroma, and the expression of c-fos as a pain marker in the fourth lumbar spinal cord were evaluated at 8 weeks postoperatively. Results The control group presented more neuroma-like features in all the observed parameters in comparison with the 2 capping groups; of the 2 capping groups, the aligned group achieved even better outcomes than the nonaligned group. Conclusions Our findings indicate that the aligned nanofiber conduit is a promising biomaterial for the nerve capping technique, and new treatment strategies using aligned nanofiber conduits may be developed for the management of painful amputated neuromas.

Extracellular high mobility group box-1 protein (HMGB1) plays important roles in the pathogenesis of nerve injury- and cancer-induced pain. However, the involvement of spinal HMGB1 in arthritis-induced pain has not been examined previously and is the focus of this study. Immunohistochemistry showed that HMGB1 is expressed in neurons and glial cells in the spinal cord. Subsequent to induction of collagen antibody-induced arthritis (CAIA), Hmgb1 mRNA and extranuclear protein levels were significantly increased in the lumbar spinal cord. Intrathecal (i.t.) injection of a neutralizing anti-HMGB1 monoclonal antibody or recombinant HMGB1 box A peptide (Abox), which each prevent extracellular HMGB1 activities, reversed CAIA-induced mechanical hypersensitivity. This occurred during ongoing joint inflammation as well as during the postinflammatory phase, indicating that spinal HMGB1 has an important function in nociception persisting beyond episodes of joint inflammation. Importantly, only HMGB1 in its partially oxidized isoform (disulfide HMGB1), which activates toll-like receptor 4 (TLR4), but not in its fully reduced or fully oxidized isoforms, evoked mechanical hypersensitivity upon i.t. injection. Interestingly, although both male and female mice developed mechanical hypersensitivity in response to i.t. HMGB1, female mice recovered faster. Furthermore, the pro-nociceptive effect of i.t. injection of HMGB1 persisted in Tlr2- and Rage-, but was absent in Tlr4-deficient mice. The same pattern was observed for HMGB1-induced spinal microglia and astrocyte activation and cytokine induction. These results demonstrate that spinal HMGB1 contributes to nociceptive signal transmission via activation of TLR4 and point to disulfide HMGB1 inhibition as a potential therapeutic strategy in treatment of chronic inflammatory pain.

Supplemental Digital Content is Available in the Text., Introduction: Paclitaxel-induced peripheral neuropathy (PIPN) is a common dose-limiting side effect of this cancer treatment drug. Spinal cord stimulation (SCS) has demonstrated efficacy for attenuating some neuropathic pain conditions. Objective: We aim to examine the inhibitory effect of SCS on the development of PIPN pain and changes of gene expression in the spinal cord in male rats after SCS. Methods: We examined whether traditional SCS (50 Hz, 6–8 h/session daily for 14 consecutive days) administered during paclitaxel treatment (1.5 mg/kg, i.p.) attenuates PIPN-related pain behavior. After SCS treatment, we performed RNA-seq of the lumbar spinal cord to examine which genes are differentially expressed after PIPN with and without SCS. Results: Compared to rats treated with paclitaxel alone (n = 7) or sham SCS (n = 6), SCS treatment (n = 11) significantly inhibited the development of paclitaxel-induced mechanical and cold hypersensitivity, without altering open-field exploratory behavior. RNA-seq showed that SCS induced upregulation of 836 genes and downregulation of 230 genes in the spinal cord of paclitaxel-treated rats (n = 3) as compared to sham SCS (n = 5). Spinal cord stimulation upregulated immune responses in paclitaxel-treated rats, including transcription of astrocyte- and microglial-related genes, but repressed transcription of multiple gene networks associated with synapse transmission, neuron projection development, γ-aminobutyric acid reuptake, and neuronal plasticity. Conclusion: Our findings suggest that traditional SCS may attenuate the development of pain-related behaviors in PIPN rats, possibly by causing aggregate inhibition of synaptic plasticity through upregulation and downregulation of gene networks in the spinal cord.

The investigators previously found that the administration of lemnalol, a natural marine compound isolated from the Formosan soft coral Lemnalia cervicorni, produced anti-inflammatory and analgesic effects in carrageenan-injected rats. Recently, several studies have demonstrated that the development and maintenance of neuropathic pain are accompanied by releasing of proinflammatory mediators from activated glial cells in the spinal cord. In this study, we investigated the antinociceptive properties of lemnalol, a potential anti-inflammatory compound, on chronic constriction injury (CCI) in a well-established rat model of neuropathic pain. Our results demonstrated that a single intrathecal administration of lemnalol (0.05-10 μg) significantly attenuated CCI-induced thermal hyperalgesia and mechanical allodynia, 14 days postsurgery. Furthermore, immunohistofluorescence analyses showed that lemnalol (10 μg) also significantly inhibits CCI-induced upregulation of microglial and astrocytic immunohistochemical activation markers in the dorsal horn of the lumbar spinal cord. Double immunofluorescent staining demonstrated that intrathecal injection of lemnalol (10 μg) markedly inhibited spinal proinflammatory mediator tumor necrosis factor-α expression in microglial cells and astrocytes in neuropathic rats. Collectively, our results indicate that lemnalol is a potential therapeutic agent for neuropathic pain, and that further exploration of the effects of lemnalol on glial proinflammatory responses is warranted.

The potential for harmful effects of general anesthesia on the developing brain has been described as pediatric anesthesiology’s greatest current challenge.1 Anesthetic agents appear to have the greatest neurodegenerative impact when exposure occurs during the period of rapid synaptogenesis.2-15 Among species, peak synaptogenesis occurs at different rates and at different parts of the life cycle. This critical period of synaptogenesis in the rat lasts from roughly day of birth to day 10, peaking in cortex at day 7. In humans, this corresponds roughly to a period from the third trimester of gestation and the initial 3 yr of postnatal life. In rats and mice, brain growth is most rapid during the first 2 weeks of postnatal life. The brain’s maturational state in postnatal day 7 mice is comparable to 7-day-old rats and corresponds to premature human neonates.16,17 Studies in infant rat models have shown that exposure to a number of classes of sedative-hypnotic medications, including benzodiazepines, n-methyl-d-aspartate receptor antagonists, and inhalation anesthetics produce striking degrees of apoptotic neurodegeneration, particularly at 7 days postnatal age (P7) but not at postnatal days 21 (P21). Similar concerns have been raised by studies in other species, including infant mice young primates and in brain slice preparations in vitro.2-15 Conversely, other investigators, including one of us (S.S.), have found increased brain apoptosis in sections of parietal and temporal cortex and thalamus in P7 rats with prolonged (6-h) ketamine exposure, but not with brief (1-h) exposure.4,18 During programmed cell death, also termed apoptosis, a cascade of autodigestive enzymes known as caspases are activated. Cleavage of the final member of this cascade, caspase-3, will leads to activation of endonucleases, resulting in apoptotic cell death.19 Case-control studies have shown that infants undergoing surgery under general anesthesia, especially with more than one procedure20 may have a higher frequency of neurodevelopmental difficulties than infants who did not require anesthesia in infancy.20 Conversely, a recent twin study did not find evidence for harmful sequelae of anesthesia in infancy.21 Case-control studies do not fully answer the question of whether the anesthesia per se is associated with increased risk, due to the confounding effect of surgery, postsurgical stress, and other comorbidities that may be more common among infants requiring surgery. To address these concerns, a large international randomized controlled trial is in progress examining neurodevelopmental outcomes at 2 and 5 years of age in a group of infants undergoing inguinal hernia repairs under spinal or general anesthesia.22 The rationale for this trial is based in part on the assumption that spinal anesthesia would not be expected to produce harmful effects on neurodevelopment. Nevertheless, this assumption has not previously been examined in infant animal surrogate models. The present study describes a technique for spinal anesthesia in the young rat. Our general hypothesis is that spinal anesthesia is neurologically benign in young rats. In particular, we hypothesized that (1): a spinal bupivacaine dose could be identified that produced signs of lower body blockade without signs of high spinal blockade, severe physiologic instability, or high mortality, (2) these doses would not increase apoptotic neurodegeneration in the brain or spinal cord (as assayed by cleaved caspase-3 staining), as seen with prolonged exposure to a volatile anesthetic, (3) spinal anesthesia would not produce long-term deficits in motor performance in those rats when tested as adults, and (4) histologic effects of spinal anesthesia on lumbar spinal cord would appear benign.

Background The authors compared the neuroprotective effects induced by two ischemic postconditioning methods and sought to determine the roles of phosphatidylinositol 3-kinase-Akt and extracellular signal-regulated kinase (ERK) in this neuroprotection. Methods Spinal cord ischemia was induced in rabbits by occlusion of the infrarenal aorta with a balloon catheter for 25 min. Postconditioning was accomplished by either five cycles of 1-min occlusion and 1-min reperfusion (standard postconditioning) or control of the perfusion pressure between 45 and 55 mmHg at the first 10 min of reperfusion (modified postconditioning). Motor function was assessed with the Tarlov score during a 28-day observation period. Histologic examination of lumbar spinal cords was performed. Expressions of Akt and ERK in the spinal cord were evaluated by Western blot. Results Compared with the controls, the two postconditioning methods markedly increased Tarlov scores 1, 3, 7, and 28 days after spinal cord ischemia and number of intact motor neurons in the lumbar spinal cord. No significant difference in Tarlov scores and number of intact motor neurons was detected between the two postconditioning method groups. The two postconditioning methods enhanced the expressions of phospho-Akt and phospho-ERK in spinal cords. The neuroprotective effects and the increases in phospho-Akt and phospho-ERK were abolished by administration of phosphatidylinositol 3-kinase-Akt inhibitor LY-294002 or ERK inhibitor PD-98059. Conclusions The two postconditioning methods possess comparable neuroprotective effects on the spinal cord and share a common molecular mechanism, in which phosphatidylinositol 3-kinase and ERK pathways play crucial roles.

BACKGROUND: Nociceptive behaviors might attenuate pain sensation. Phosphorylation of extracellular signal-regulated kinase (pERK) was recently reported to be induced by noxious stimuli in dorsal horn neurons. We investigated, in a formalin test, whether pERK of the dorsal horn is affected by licking. METHODS: Twenty-four adult male rats were divided into four groups: control, formalin test, restricted control, and restricted formalin test. Ten percent formalin was injected subcutaneously into the left rear paw of the formalin test and restricted formalin test groups. The control and formalin test group rats were kept in a clear plastic chamber, whereas the restricted control and restricted formalin test group rats were kept in a modified-restraint, pipe-shaped chamber. All rats were killed after 25 min. Twelve sections of the lumbar spinal cord were processed for p-ERK immunohistochemistry using the avidin-biotin peroxidase method. RESULTS: The number of p-ERK positive cells in the restricted formalin test group was significantly higher than in the other three groups in the ipsilateral-side superficial dorsal horn (P < 0.05). However, there was no significant difference between the formalin test group and the two control groups in pERK expression. CONCLUSION: Licking decreased pERK of the spinal cord of the formalin test group. The findings suggested that licking attenuated the pain of the formalin test.

The duration of noxious stimulus-induced antinociception (NSIA) has been shown to outlast the pain stimulus that elicited it, however, the mechanism that determines the duration of analgesia is unknown. We evaluated the role of spinal excitatory and inhibitory receptors (NMDA, mGluR(5), mu-opioid, GABA(A), and GABA(B)), previously implicated in NSIA initiation, in its maintenance. As in our previous studies, the supraspinal trigeminal jaw-opening reflex (JOR) in the rat was used for nociceptive testing because of its remoteness from the region of drug application, the lumbar spinal cord. NSIA was reversed by antagonists for two inhibitory receptors (GABA(B) and mu-opioid) but not by antagonists for either of the two excitatory receptors (NMDA and mGluR(5)), indicating that NSIA is maintained by ongoing activity at inhibitory synapses in the spinal cord. Furthermore, spinal administration of the GABA(B) agonist baclofen mimicked NSIA in that it could be blocked by prior injection of the mu-opioid receptor antagonist H-D-Phe-Cys-Tyr-D-Trp-Arg-Thr-Pen-Thr-NH(2) (CTAP) in nucleus accumbens. CTAP also blocked baclofen antinociception when administered in the spinal cord. We conclude that analgesia induced by noxious stimulation is maintained by activity in spinal inhibitory receptors.

Study design Case report. Objectives To report a rare case of complicated paraplegia caused by a spontaneous epidural hematoma following an epidural steroid injection in a patient with unrecognized chronic idiopathic thrombocytopenic purpura (ITP) and to review relevant literature and discuss etiology, pathogenesis, and clinical features. Summary of background data A spinal epidural hematoma is a rare but potentially catastrophic complication, which could develop in patients without any risk factors. Some patients with chronic ITP are asymptomatic. To our knowledge there has been no previous report of such a complication. Methods This is a retrospective review of a case seen at our institution. Results The authors present a case of a 67-year-old woman who received an epidural steroid injection following complaints of lower back pain and bilateral buttock and leg pain. One day later, the patient had right leg numbness and weakness extending to her right knee; she was taken for emergency surgery. An emergency magnetic resonance imaging revealed an epidural hematoma with high-signal intensity on T2 imaging in the lumbar spinal cord and spinal cord compression with subdural hematoma. One week later, she was progressively developing lower extremity paraplegia with an L1 motor level and no sensory or sphincter activity. She was taken from the emergency room at our institution to the operating room for emergency decompression. After an uneventful course for 1 year, the patient presented with progressive bilateral lower extremity paralysis. Conclusion Epidural steroid therapy is a commonly used conservative therapy; however, complications could develop in patients without any risk factors. Clinicians who plan an epidural steroid injection must perform a rigorous evaluation through a detailed physical examination, simple laboratory tests, and history taking to prevent various risks associated with spinal cord compression.

BACKGROUND: Using a rat L5 spinal nerve transection model we previously showed that pentoxifylline prevents hyperalgesia through antiinflammation in the prefrontal brain. In this study, we examined efficacy when applied after injury. METHODS: We examined the effect of pentoxifylline on existing mechanical allodynia, observing glial activation and proinflammatory cytokine expression in the lumbar spinal cord, when given 7 days after L5 spinal nerve transection. RESULTS: There was no effect from pentoxifylline on existing hypersensitivity, glial activation, and cytokine expression when applied after L5 spinal nerve transection. CONCLUSION: Pentoxifylline administered intraperitoneally on day 7 postsurgery failed to alleviate existing hypersensitivity, or reduce glial activation and cytokine expression.

CGRP receptor activation has been implicated in peripheral and central sensitization. The role of spinal CGRP receptors in supraspinal pain processing and higher integrated pain behavior is not known. Here we studied the effect of spinal inhibition of CGRP1 receptors on supraspinally organized vocalizations and activity of amygdala neurons. Our previous studies showed that pain-related audible and ultrasonic vocalizations are modulated by the central nucleus of the amygdala (CeA). Vocalizations in the audible and ultrasonic range and hindlimb withdrawal thresholds were measured in awake adult rats before and 5–6 h after induction of arthritis by intra-articular injections of kaolin and carrageenan into one knee. Extracellular single-unit recordings were made from neurons in the latero-capsular division of the CeA (CeLC) in anesthetized rats before and after arthritis induction. CGRP1 receptor antagonists were applied to the lumbar spinal cord intrathecally (5 μl/min) 6 h postinduction of arthritis. Spinal administration of peptide (CGRP8-37, 1 μM) and non-peptide (BIBN4096BS, 1 μM) CGRP1 receptor antagonists significantly inhibited the increased responses of CeLC neurons to mechanical stimulation of the arthritic knee but had no effect under normal conditions. In arthritic rats, the antagonists also inhibited the audible and ultrasonic components of vocalizations evoked by noxious stimuli and increased the threshold of hindlimb withdrawal reflexes. The antagonists had no effect on vocalizations and spinal reflexes in normal rats. These data suggest that spinal CGRP1 receptors are not only important for spinal pain mechanisms but also contribute significantly to the transmission of nociceptive information to the amygdala and to higher integrated behavior.

Background Spinal prostaglandins are important in the early pathogenesis of spinal nerve ligation (SNL)-induced allodynia. This study examined the effect of SNL on the expression of cyclooxygenase (COX)-1, COX-2, and prostaglandin E2 receptors in the rat lumbar spinal cord, and the temporal and pharmacologic relation of these changes to the exaggerated A- and C-fiber-mediated reflex responses and allodynia, 24 h after injury. Methods Male Sprague-Dawley rats, fitted with intrathecal catheters, underwent SNL or sham surgery. Paw withdrawal threshold, electromyographic analysis of the biceps femoris flexor reflex, and immunoblotting of the spinal cord were used. Results Both allodynia (paw withdrawal threshold of < or = 4 g) and exaggerated A- and C-fiber-mediated reflex responses (i.e., decrease in activation threshold, increase in evoked activity, including windup; P < 0.05) were evident 24 h after SNL but not sham surgery. Allodynic animals exhibited significant increases in prostaglandin E2 receptor (subtypes 1-3) and COX-1 (but not COX-2) expression in the ipsilateral lumbar dorsal horn. The corresponding ventral horns and contralateral dorsal horn were unchanged from sham controls. Exaggerated A- and C-fiber-mediated reflex responses were significantly attenuated by intrathecal SC-560 or SC-51322, but not SC-236, given 24 h after SNL. Conclusion These results provide further evidence that spinal prostaglandins, derived primarily from COX-1, are critical in the exaggeration of A- and C-fiber input and allodynia, 24 h after SNL.

BACKGROUND: Because of the logistical obstacles to measurement under hyperbaric conditions, the effect of nitrous oxide (N 2 O) alone on spinal neuronal responses has not been tested. We hypothesized that, like other inhaled anesthetics, N 2 O would depress spinal neuronal responses to noxious stimulation. METHODS: The lumbar spinal cord was exposed in rats anesthetized with isoflurane. Mechanically ventilated rats were placed into a hyperbaric chamber and needle electrodes were inserted into the hindpaws. Isoflurane administration was discontinued and anesthesia converted to N 2 O by pressurizing the chamber with N 2 O. A microelectrode was inserted into the lumbar cord using computer-controlled motors and a hydraulic microdrive. Neuronal responses to electrical stimulation of the hindpaw were sought at 1.5, 2, and 2.5 atm N 2 O (0.8-1.3 minimum alveolar concentration). RESULTS: Increasing N 2 O partial pressures variably affected neuronal responses to a 2 s 100-Hz electrical stimulus. Neuronal depth and neuronal response were correlated, with superficial neurons tending to be facilitated, while deeper neurons were depressed; (overall responses were 1331 ± 408, 1594 ± 383, and 1578 ± 500 impulses/min at 1.5, 2, and 2.5 atm N 2 O, respectively; mean, standard error). N 2 O did not affect neuronal responses to a repetitive "windup" stimulus. Infusion of the N-methyl-D-aspartate blocker MK-801 into separate rats increased the neuronal response to the 100-Hz stimulus (from 781 ± 216 to 1352 ± 269 impulses/min, P < 0.05). CONCLUSIONS: N 2 O facilitated superficial spinal neuronal responses to noxious stimulation while depressing deeper neurons. These results suggest that anesthetic partial pressures of N 2 O have divergent effects on spinal neuronal responses to noxious stimulation, the specific responses depending on the depth of the spinal neurons.

Intense pain or intense peripheral inflammation experienced during development can have pronounced effects upon adult pain sensation. However, little is known about the more commonly encountered mild systemic inflammation, such as that experienced with mild illness. Neonatal exposure to lipopolysaccharide (LPS), an established model of immune system activation, has been shown to affect febrile and cyclooxygenase-2 (COX-2) responses to a similar exposure in adulthood. Adult LPS also elicits a range of sickness behaviours, including enhanced responses to painful stimuli. We, therefore, hypothesized that adult sensation and pain responses could be affected by a neonatal LPS challenge. Male and female Sprague-Dawley rats were administered LPS at postnatal day 14 and were tested in adulthood for nociceptive responses to thermal and mechanical stimuli using, respectively, a plantar test apparatus and von Frey filaments, before and after adult LPS. Expression of dorsal root ganglion and lumbar spinal cord COX-2 was also examined. Animals treated as neonates with saline showed the expected hypersensitivity to painful stimuli after adult LPS as well as enhanced spinal cord COX-2. Neonatally LPS-treated rats, however, showed a significantly different profile. They displayed enhanced baseline nociception and elevated basal spinal cord COX-2 compared with neonatally saline-treated rats. Also, rather than the expected hyperalgesia after adult LPS, no changes in nociceptive responses and a reduction in spinal cord COX-2 expression were observed. These findings have important implications for the understanding of pain and its management and highlight the importance of the neonatal period in the development of pain pathways.

We investigated the effects of IV midazolam on spinal cord blood flow in 32 cats anesthetized with isoflurane. Cats underwent laminectomy, and the lumbar spinal cord was exposed. A platinum electrode was inserted stereotaxically into the spinal cord to a depth of 1 mm-2 mm lateral to midline at L2. Arterial blood pressure, heart rate, and spinal cord blood flow (using the hydrogen clearance method) were measured before and at 5, 15, 30, 60, 90, and 120 min after an IV bolus of midazolam (0, 1, 2, or 4 mg/kg in saline 5 mL; n = 8 cats per dose). Arterial blood pressure was not affected by 0 or 1 mg/kg of midazolam but was decreased for 30 min by 2 or 4 mg/kg of midazolam. Heart rate did not change. Spinal cord blood flow was increased for 90 min by midazolam 1 mg/kg and for 15 min by midazolam 2 mg/kg but was not changed by midazolam 4 mg/kg. In conclusion, 1 mg/kg of midazolam increased feline spinal cord blood flow without changing arterial blood pressure. In contrast, a larger dose of midazolam (4 mg/kg) did not change spinal cord blood flow but substantially decreased arterial blood pressure during isoflurane anesthesia.

PURPOSE OF REVIEW Subarachnoid injection of local anesthetics has been related to the appearance of transient neurological symptoms (called transient neurologic syndrome), as reflected by a number of clinical reports showing their incidence in clinical practice. However, the etiology of this syndrome is virtually unknown, as is the number of factors implicated in its development. This review will attempt to clarify this entity and its relationship with spinal anesthesia. RECENT FINDINGS Intrathecal administration of local anesthetics is known to increase glutamate concentration in cerebrospinal fluid and histopathologic changes of motor neurons in the lumbar spinal cord, suggesting damage of dorsal and ventral roots. In-vitro studies of cultured neurons exposed to different concentrations of local anesthetics have shown changes in growth of cones and neurites, which may be related to transient neurologic syndrome. SUMMARY The latest studies show biochemical and anatomopathologic changes that support the structural basis of the existence of transient neurologic syndrome. In the authors' view, transient neurologic syndrome could represent the lower end of a spectrum of local anesthetic toxicity. Recent findings demonstrate that nerve membrane damage induced by highly concentrated local anesthetics such as lidocaine, tetracaine, dibucaine and procaine may generate irreversible neural injury. Still further studies are needed to establish the relationship between morphological changes induced in vitro and the occurrence of clinical symptoms.

The stress-activated mitogen-activated protein kinase (MAPK) p38 is emerging as an important mediator of pain. The present study examined the possible involvement of peripheral and spinal p38 MAPK in capsaicin-induced thermal hyperalgesia. Topical capsaicin produced phosphorylation of p38 MAPK in the skin from the affected hindpaw as well as the corresponding lumbar spinal cord in a time dependent manner. Topical capsaicin produced robust C-fiber mediated thermal hyperalgesia that was inhibited by systemic, local peripheral, or central intrathecal pre-treatment with the p38 MAPK inhibitor, SD-282. Intraperitoneal SD-282 (10-60 mg/kg) significantly and dose-dependently attenuated capsaicin-induced C-fiber mediated thermal hyperalgesia. Similarly, 0.1-5mg/kg subcutaneous SD-282 in the hindpaw dose-dependently attenuated capsaicin-induced thermal hyperalgesia. Intrathecal administration of 1microg SD-282 was also anti-hyperalgesic in this model. Functionally, SD-282 decreased capsaicin-induced release of calcitonin gene related peptide in an in vitro skin release assay, consistent with a role for p38 MAPK in peripheral nerve function. These results suggest that p38 MAPK plays a role in the development of hyperalgesic states, exerting effects both centrally in the spinal cord and peripherally in sensory C fibers.

Tissue damage during surgery can induce 'central sensitization' and the development of pain and hyperalgesia post-operatively. Metabotropic glutamate receptors (mGluRs) contribute to nociception, inflammatory pain and hyperalgesia. This study characterized the temporal expression of group I (mGluR(1), mGluR(5)) and II (mGluR(2), mGluR(3)) mGluRs in spinal cord following abdominal surgery. Lumbar spinal cord was recovered from adult sheep euthanased 5 h, 1, 2, 3 and 6 days after undergoing a midline laparotomy, and processed for mGluR mRNA (real-time PCR, in situ hybridization) and protein (Western blotting). mGluR(5) mRNA was up-regulated 5 h and 1 day post-surgery in laminae I-II of the spinal cord dorsal horn. mGluR(5) protein was increased 1 day post-surgery. A delayed induction of mGluR(2) and mGluR(3) mRNAs and mGluR(2/3) protein occurred in spinal cord 3 days after surgery. By 6 days, mGluR(2) mRNA levels had returned to normal, however, mGluR(3) mRNA and mGluR(2/3) protein remained elevated. No change was detected in mGluR(1). These results demonstrate that mGluRs are differentially regulated following surgery and support a link between mGluR-mediated activity and post-surgical pain.

We examined ultrastructural details such as the cellular component and membrane thickness of human spinal pia mater with the aim of determining whether fenestrations are present. We hypothesized that pia mater is not a continuous membrane but, instead, that there are fenestrations across the pial cellular membrane. The lumbar dural sac from 7 fresh human cadavers was removed, and samples from lumbar spinal pia mater were studied by special staining techniques, immunohistochemistry, and transmission and scanning electron microscopy. A pial layer made by flat overlapping cells and subpial tissue was identified. We found fenestrations in samples from human spinal pia mater at the thoracic-lumbar junction, conus medullaris, and nerve root levels, but these fenestrations did not appear at the thoracic level. We speculate whether the presence of fenestrations in human spinal pia mater at the level of the lumbar spinal cord and at the nerve root levels has any influence on the transfer of local anesthetics across this membrane.The ultrastructural anatomy of the human pia mater, such as pial cells, membrane thickness, and subpial tissue at different levels of the thoracic and lumbar spinal cord and nerve roots, was studied by special staining techniques, immunohistochemistry, and transmission and scanning electron microscopy. Fenestrations were found in samples at the thoracic-lumbar junction, conus medullaris, and nerve root levels. No fenestrations were found in samples at the thoracic level. At present, we cannot determine the significance of these findings.

Objective: To explore the feasibility of performing percutaneous endoscopic cellular transplantation into the lumbar spinal cord of pigs to create intramedullary cellular trails. METHODS: The lumbar subarachnoid space was accessed using a 10-gauge needle inserted between L5 and L6. A 12.5-French flexible introducer sheath was fed over the needle into the subarachnoid space. A 3.2-mm-diameter flexible, steerable endoscope was then directed intadurally through the sheath. The thecal space was distended by saline infusion. A microcatheter with an attached needle then was advanced through the working channel into the dorsal surface of the lumbar spinal cord. Five microliters of Hoechst-labeled fibroblasts were injected while the catheter was withdrawn slowly to create a trail of cells within the spinal cord. The spinal canal then was perfused with fixative. The injected spinal cord segment was removed and studied histologically. Endoscopic video was analyzed offline. RESULTS: The endoscope could be navigated under visual guidance. The sacral and lumbar rootlets, the spinal cord, and associated vessels were visualized. In fixed sagittal sections, a linear trail of fluorescent fibroblasts could be seen within the lumbar spinal cord in earch specimen. CONCLUSION: Percutaneous endoscopic cellular injection may be usegul for cellular transplantation, may reduce surgical and anesthetic time, may be compatible with local anesthesia, may eliminate the need to disrupt spinal instrumentation and bone grafts, and may allow greater flexibility in the respective timing of spinal fixation and cellular transplantation after spinal cord injury. This is the first report of the use of endoscopi intraspinal cellular transplantation.

Background Although upregulation of cyclooxygenase (COX)-2 in spinal cord after peripheral inflammation has been well documented, the effect of surgery on spinal COX-2 has not been examined in detail. The present study uses a bilateral foot incision in rats to examine the magnitude and duration of surgically induced changes in spinal COX-2 protein. Methods A longitudinal incision was made in both plantar hind paws of isoflurane-anesthetized rats. Spinal cords were removed at various postoperative times (1-48 h), and spinal COX-2 protein levels were compared with the results of Western blot analysis. Ropivacaine-induced blockade of sciatic nerve function was used to determine the importance of afferent nerve activity on spinal COX-2 after incision. Dexamethasone and the COX-2-selective inhibitor L-745,337 were administered intrathecally to modulate spinal COX-2 after incision. Results COX-2 protein levels increased in the lumbar spinal cord at 3 (1.32-fold) and 6 (1.26-fold) h after bilateral foot incision. At later times, lumbar COX-2 levels were no different than in control animals not undergoing surgery. Cervical COX-2 protein levels remained unchanged. Sciatic nerve blockade with ropivacaine did not prevent the increase in lumbar spinal COX-2 protein levels after incision. Intrathecal dexamethasone decreased lumbar spinal COX-2 levels after incision, and an intrathecal COX-2-selective inhibitor did not reduce the COX-2 upregulation. Conclusions After bilateral foot incision in rats, lumbar spinal COX-2 protein levels increase, although the magnitude and duration are less than reported in models of peripheral inflammation. This COX-2 upregulation does not seem to be mediated by afferent nerve activity.

We investigated the influence of gravity on the maturation of electrical properties of lumbar motoneurons and the development of their inputs from ventral descending pathways, which are important for the control of posture and locomotion. Using electrophysiological approaches in the in vitro brain stem-spinal cord preparation of neonatal rats born and reared in hypergravity field we demonstrate that: (1) the postnatal development of descending inputs to lumbar enlargement was reduced in animals submitted to hypergravity; (2) similar developmental pattern of basic electrical properties observed between motoneurons of hypergravity and control animals could not account for the changes in descending inputs. We concluded that gravity was critical to shape development of the supraspinal afferents in the lumbar spinal cord throughout the postnatal period.

UNLABELLED To investigate the analgesic effects of xenon, we performed formalin tests in rats under 0.5 minimum alveolar anesthetic concentration xenon or nitrous oxide and stained the lumbar spinal cord for c-fos (n = 18) and the phosphorylated N-methyl-D-aspartate (NMDA) receptor (n = 24) by using the avidin-biotin-peroxidase method. After 20 min of 79% xenon, 68% nitrous oxide, or 100% inhaled oxygen, 10% formalin (100 microL) was injected into the left rear paw of the animals except for a control group. Nociceptive behavior was observed for 1 h. The rats were killed 2 h after the formalin injection, and the lumbar spinal cord was stained for c-fos or the phosphorylated NMDA receptor immunohistochemically. Animals in the xenon and nitrous oxide groups showed less nociceptive behavior than did the oxygen group. Although the number of c-fos-positive cells in the lumbar spinal cord in the nitrous oxide group was not decreased, that in the xenon group decreased. The number of phosphorylated NMDA receptor-positive cells in the xenon group was significantly less than in the nitrous oxide and oxygen groups. Inhaled xenon suppressed nociceptive behaviors, c-fos expression, and activation of the NMDA receptor during the formalin test in rats. These results confirm that xenon's analgesic effects result from inhibition of the NMDA receptor. IMPLICATIONS Inhaled xenon suppressed nociceptive behaviors, c-fos expression, and activation of the N-methyl-D-aspartate receptor during the formalin test in rats. Xenon's analgesic effect was speculated to result from the inhibition of the N-methyl-D-aspartate receptor in vivo.

Recent studies demonstrate the possible existence of tonic modulatory control of nociceptive input mediated by spinal cannabinoid receptors (CB1). Accordingly, it is predicted that a reduction in the spinal CB1 receptors may enhance sensitivity to sensory stimuli and a decrease in spinal antinociceptive potency to cannabinoid agonists. An antisense oligodeoxynucleotide (ODN) specific to the CB1 receptor was used to 'knock-down' CB1 receptors in the lumbar spinal cord and dorsal root ganglia by the local, repeated intrathecal (i.th.) administration of the ODN. This treatment resulted in a decrease in lumbar spinal CB1 receptor expression accompanied by a decrease in the response thresholds to both innocuous tactile and noxious thermal stimuli. The antinociceptive action of the CB1 agonist, WIN 55,212-2, by i.th. administration was also significantly attenuated after treatment with the antisense ODN. Similar treatment using a mismatch control ODN had no effect on receptor protein or on sensory thresholds. The effects of the antisense ODN treatment on sensory thresholds were fully reversed after discontinuation of the ODN injection. The antisense ODN treated rats also showed a significant increase in lumbar spinal dynorphin A. Acute i.th. injection of MK-801 or an antidynorphin antiserum blocked the antisense ODN-induced tactile and thermal hypersensitivity. These data support the possibility of endogenous inhibitory cannabinoid tone to limit spinal afferent input of thermal and tactile stimuli. Lifting of this inhibitory tone through a 'knock-down' of spinal CB1 receptors apparently lowers the thresholds for sensory input, as reflected by the actions of MK-801 to block tactile and thermal hypersensitivity. The increased spinal dynorphin may act to further promote afferent outflow and abnormal pain because sequestration of spinal dynorphin with antiserum also reverses the manifestations of abnormal pain following knock-down of CB1 receptors.

The present study was undertaken to determine whether leukocytes are recruited into the spinal cord following a peripheral L5 spinal nerve transection that results in mechanical allodynia (increased tactile sensitivity behavior correlates with neuropathic pain). In rats subjected to bone marrow irradiation, donor-specific major histocompatibility complex (MHC) class I (I1-69) positive peripheral immune cells trafficked to the L5 spinal cord in response to an L5 spinal nerve injury. The number of I1-69 positive cell profiles increased over time and correlated with increased mechanical allodynia. At early time points following injury, I1-69 positive immune cells co-regionalized with the expression of the macrophage marker ED2. At later time points following injury, some of the infiltrating immune cells did not co-regionalize with the macrophage marker ED2. At no time did the infiltrating cells co-regionalize with the neuronal marker (NeuN). Both macrophage-like morphology and T cell-like morphology were observed in the I1-69 positive cellular infiltrate. Conversely, animals that underwent sham surgery demonstrated little mechanical allodynia and a minimal number of infiltrating peripheral immune cells. In a separate group of rats, infiltration of CD3+ T-lymphocytes was confirmed at 14 days post-nerve transection. This study demonstrates trafficking of leukocytes into the lumbar spinal cord at time points that correlate with mechanical allodynia suggesting a role of central neuroinflammation in persistent neuropathic pain.

Background Nerve injury in animals increases alpha(2)-adrenoceptor expression in dorsal root ganglion cells and results in novel excitatory responses to their activation, perhaps leading to the phenomenon of sympathetically maintained pain. In contrast to this notion, peripheral alpha(2)-adrenoceptor stimulation fails to induce pain in patients with chronic pain. We hypothesized that alpha(2) adrenoceptors at the site of nerve injury play an inhibitory, not excitatory role. Methods Partial sciatic nerve ligation was performed on rats, resulting in a reduction in withdrawal threshold to tactile stimulation. Animals received perineural injection at the injury site of clonidine, saline, or clonidine plus an alpha(2)-adrenergic antagonist, and withdrawal threshold was monitored. Immunohistochemistry was performed on the sciatic nerve ipsi- and contralateral to injury and on the spinal cord. Results Clonidine reduced this hypersensitivity in a dose-dependent manner, and this was blocked by an alpha(2A)-preferring antagonist. Perineural clonidine injection had a slow onset (days) and prolonged duration (weeks). Systemic or intrathecal clonidine, or transient neural blockade with ropivacaine, had short lasting or no effect on hypersensitivity. alpha(2A)-adrenoceptor immunostaining was increased near the site of peripheral nerve injury, both in neurons and in immune cells (macrophages and T lymphocytes). Phosphorylated cAMP response element binding protein (pCREB) in lumbar spinal cord was increased ipsilateral to nerve injury, and this was reduced 1 week after perineural clonidine injection. Conclusions These data suggest that peripheral alpha(2) adrenoceptors are concentrated at the site of peripheral nerve injury, and their activation receptors produce long-lasting reductions in abnormal spinal cord gene activation and mechanical hypersensitivity.

We examined the effects of systemically, spinally, and peripherally administered amitriptyline on formalin-induced Fos immunoreactivity in the lumbar spinal cord. Formalin (2.5%), injected subcutaneously into the rat hindpaw, increased Fos immunoreactivity in laminae I-II, III-IV, and V-VI of the dorsal L5 spinal cord. Amitriptyline, administered both systemically and spinally before formalin, increased flinching and concurrently decreased biting/licking behaviors, but neither route of administration produced any statistically significant change in Fos immunoreactivity. Amitriptyline coadministered with the formalin reduced both flinching and biting/licking behaviors, and significantly reduced Fos immunoreactivity, particularly in laminae I-II. These immunohistochemical changes reflect the net behavioral effects observed after the different routes of drug administration. The profile of amitriptyline action after peripheral administration may be of clinical importance because of the potential use of antidepressants as topical analgesics.In the formalin test, amitriptyline produces different effects on pain behaviors after systemic, spinal administration and peripheral administration. Fos protein, an indicator of neuronal activity after noxious stimulation, is upregulated after formalin injection. We examined the effects of amitriptyline on such expression and observed a reduction in expression with peripheral administration.

Background Spinally administered opioids show decreased potency and efficacy in the treatment of neuropathic pain. As reported previously, morphine stimulates spinal opioid receptors to effect adenosine release, which acts at adenosine receptors to produce analgesia. The authors hypothesized that morphine induces less adenosine release in neuropathic compared with normal rats, explaining its reduced potency and efficacy. Methods Sprague-Dawley rats (200-250 g) were divided into three groups: no surgery (n = 52), sham surgery (n = 20), or left L5 and L6 spinal nerve ligation (n = 64). Two weeks after surgery, mechanical hypersensitivity of the left hind paw was verified. For each experiment, a crude synaptosomal P2 suspension was prepared by homogenizing cervical and lumbar dorsal spinal cord halves from four rats, followed by differential centrifugation, and aliquots incubated with morphine sulfate from 10(-8) to 10(-4) m alone or in presence of 10(-5) m dipyridamole. Extrasynaptosomal concentrations of adenosine were analyzed by high-pressure liquid chromatography. Results Synaptosomal release of adenosine in the absence of morphine was similar between groups. Morphine produced a concentration-dependent adenosine release, which was less in synaptosomes from dorsal lumbar spinal cord in spinal nerve ligation compared with normal or sham animals. This reduction was removed by adding dipyridamole. Conclusion Morphine normally stimulates spinal release of adenosine, a potent antihypersensitivity compound. Because this effect of morphine is diminished in spinal nerve ligation animals, one explanation for decreased efficacy and potency of opioids in the treatment of neuropathic pain may be a dipyridamole-sensitive disruption in the opioid-adenosine link in the spinal cord.

We studied the role of glutamate excitotoxicity in motor neuron degeneration in the wobbler mouse (wr/wr), a model of amyotrophic lateral sclerosis and spinal muscular atrophies. Choline acetyltransferase (ChAT) activity was decreased in the cervical spinal cord and in the muscles innervated by nerves originating in this region of wobbler mice, but no differences were found in the lumbar spinal cord and in the hindleg muscles. Glial fibrillar acid protein (GFAP), a marker of reactive gliosis, was significantly higher in the cervical spinal cord of wobbler mice aged 4 weeks than in controls and the differences were more marked at 12 weeks; no differences were found in the lumbar spinal cord. In spite of this selective degeneration of motor neurons (resulting in strong decrease in the neuronal glutamate transporter EAAC1) and reactive gliosis in the cervical spinal cord, the levels of the glial glutamate transporter proteins GLT-1 and GLAST were similar in wobbler and control mice. Plasma concentrations of excitatory amino acids were no different at any time examined. Our results exclude the involvement of decrease in glutamate GLT 1 transporter in the motor neuron degeneration in wobbler mice.

Using immunohistochemistry and in situ hybridization the expression of calcitonin gene-related peptide (CGRP) was studied in the mouse spinal cord under normal conditions and after unilateral rhizotomy and after local colchicine treatment. Under normal conditions a dense plexus of CGRP-immunoreactive (IR) fibres was observed in the superfical layers of dorsal horns with lower numbers of fibers in deeper laminae. Seven days after unilateral rhizotomy, there was a marked reduction of CGRP-IR fibres in the ipsilateral superfical layers and distinctly CGRP-IR neurons could be detected in the ipsilateral lamina III. CGRP mRNA-positive neurons were observed in lamina III in both the ipsilateral and contralateral dorsal horn. Colchicine treatment did not markedly increase the number of CGRP-IR neurons. The results suggest that CGRP is synthesized in local dorsal horn neurons of the mouse, and these neurons presumably participate in sensory processing. NeuroReport 12:739-743 (C) 2001 Lippincott Williams & Wilkins.

Studies using mSOD1 transgenic mice, an animal model of familial amyotrophic lateral sclerosis (fALS), suggest that endothelial damage and impaired integrity of the blood–brain barrier (BBB) and blood–spinal cord barrier (BSCB) occur before signs of weakness and may contribute to motoneuron injury, further implicating a non-cell-autonomous pathogenesis.1–3 While tight junctions were still intact in late symptomatic mSOD1 animals, there were ultrastructural alterations in the BBB and BSCB and leakage of Evans Blue dye from spinal cord microvessels at both early and late stages of disease.1,2 A recent study also demonstrated the presence of microhemorrhages and further showed decreased expression of the tight junction proteins zona occludens 1 (ZO-1), occludin (Ocln), and claudin-5 (Cldn5) in mSOD1 mice during disease progression.3 To investigate whether tight junction proteins are decreased in patients with ALS, the mRNA expression of ZO-1, Ocln, and Cldn5 was assessed in spinal cord tissue from patients with sporadic ALS (sALS), patients with fALS, and non-neurodegenerative disease controls (NNDC). ### Methods. After receiving written consent and approval by The Methodist Hospital or the University of Pittsburgh School of Medicine, postmortem tissue samples were obtained from patients with a definite diagnosis of ALS according to WFN El Escorial/Airlie criteria. RNA was extracted from homogenized lumbar spinal cord specimens from 30 patients with sALS, 4 patients with fALS, and 16 NNDC with Trizol (Invitrogen, Grand Island, NY) and purified with an RNeasy kit (Qiagen, Valencia, CA). Quantitative reverse transcriptase PCR (RT-PCR) was performed using 10 ng of RNA, an iScript One-step RT-PCR kit with SYBR Green (Bio-Rad, Hercules, CA), and assayed on an iQ5 Multicolor …

Nociceptive C fiber input to SI in the halothane-nitrous oxide anesthetized rat was assessed by recording cortical field potentials evoked by noxious thermal cutaneous stimulation with CO2-laser pulses. Morphine topically applied onto the lumbar spinal cord produced a dose-dependent inhibition of nociceptive C fiber input from the hind paw to the contralateral SI. The inhibitory effect of morphine was reversed by naloxone. Potentials evoked by CO2-laser stimulation of the forepaw were unaffected by morphine applied on the lumbar cord, indicating that the effect of morphine was exerted at the segmental level. It is concluded that input from nociceptive C fibers to SI is relayed in the spinal cord and can be inhibited by spinal opioid receptor activation. The present method offers an interesting model of ascending nociceptive transmission to the cerebral cortex.

To investigate whether sublethal ischemia preserves neuronal function otherwise lost after stroke, anesthesized rabbits were subjected to clamping of abdominal aorta to cause lumbar spinal cord ischemia. An occlusion period of 12.5 min was followed 12 or 48 h later by a second occlusion for 30 min. When scored 24 h later for hindlimb function on a 0-6 scale, the rabbits that underwent tolerizing ischemia 12 h before infarction had better motor function (n = 7; 4.29 ± 0.21, p < 0.0001) than sham-operated controls (n = 7; 1.00 ± 0.27), but those infarcted at 48 h had mixed outcomes (n = 5; 2.20 ± 0.21, ns). In correlation, the proportion of neurons with histological evidence of damage was lower in the tolerized rabbits (0.15 ± 0.04) than in sham-operated controls (0.74 ± 0.09, p < 0.001). We conclude that ischemic tolerance also improves neurological function of infarcted spinal cord and could be studied for clinical application.

Adrenal chromaffin cells produce analgesic substances, such as catecholamines and enkephalins, and intrathecal (i.t.) implantation of either allografted adrenal tissue or xenogenic chromaffin cells produce antinociception in animals. We evaluated the analgesic effect of bovine chromaffin cells in a model of central pain in which rats exhibit chronic allodynia-like behavior after photochemically induced ischemic spinal cord injury. Bovine chromaffin cells or endothelial cells were injected i.t. onto the lumbar spinal cord and their effects on mechanical and cold allodynia-like behaviors were studied for up to 8 weeks. The chronic allodynia-like behavior was stable for months without signs of remission and i.t. implantation of human endothelial cells did not alleviate the chronic allodynia-like behavior for the entire observation period. In contrast, 2 weeks after i.t. implantation of bovine chromaffin cells, the mechanical allodynia was abolished in the spinally injured rats, and the enhanced response to cold stimuli was significantly reduced. The overall effects were significant up to 8 weeks after i.t. implantation, although the anti-allodynic effect decreased towards the end of the observation period. No signs of side-effects were noted after i.t. implantation. The allodynia-like state was temporarily restored by naloxone (0.5 mg/kg) or phentolamine (0.3 mg/kg) injected intraperitoneally. Immunohistochemical examination revealed that tyrosine hydroxylase (TH)-positive chromaffin cells could be identified adjacent to the spinal cord up to 4 weeks after i.t. implantation, whereas at 8 weeks the TH-positive cells were sparse. It is concluded that bovine chromaffin cells stay viable in rat spinal cord for a considerable period of time after i.t. administration and alleviate chronic allodynia-like behavior in spinally injured rats, possibly through activation of opioid and alpha-adrenoceptors. The present results further document a new therapeutic approach for the treatment of chronic neuropathic pain.

Background Controversy exists over the efficacy of different methods for protecting the spinal cord against experimental ischemic injury. Therefore, the authors compared the protective effects of thiopental with those of hypothermia (35 degrees C and 32 degrees C) on hindlimb motor functions and histopathology after transient spinal cord ischemia. Methods Twenty-seven New Zealand white rabbits were assigned to one of the four groups: a thiopental-normothermia group (burst-suppression dose of thiopental; esophageal temperature = 38 degrees C; n = 7), a halothane-mild hypothermia group (halothane, 1%; esophageal temperature = 35 degrees C; n = 7), a halothane-moderate hypothermia group (halothane, 1%; esophageal temperature = 32 degrees C; n = 6), and a halothane-normothermia group (halothane, 1%; esophageal temperature = 38 degrees C; n = 7). The animals were then subjected to 20 min of spinal cord ischemia produced by occlusion of the aorta distal to the origin of left renal artery. Hindlimb motor function was observed for 48 h after reperfusion. Histopathology of the lumbar spinal cord also was examined. Results All animals in the halothane-mild hypothermia and halothane-moderate hypothermia groups were neurologically normal 48 h after ischemia. There was no statistical difference in the final neurologic status and histopathology between the thiopental-normothermia and halothane-normothermia groups. However, the final neurologic status and histopathology in both groups were worse than in the halothane-mild hypothermia or halothane-moderate hypothermia groups. There was a strong correlation between the final neurologic status and the numbers of normal neurons in the anterior spinal cord. Conclusions These results suggest that mild and moderate hypothermia protects against ischemic spinal cord injury in rabbits, and a burst-suppression dose of thiopental does not offer any advantage over halothane.

We compared cerebrospinal fluid (CSF) drainage (Group D ; n = 8) to neuroanesthesia adjunct therapy (hyperventilation and mannitol administration ; Group N ; n = 8) for the prevention of paraplegia using a canine model of descending thoracic aortic cross-clamping (AXC ; 2.5 mm distal to the left subclavian artery for 30 min). We expected no difference in neurologic outcome between groups. After surgical preparation and a 30-min stabilization period, dogs in Group D had CSF drained prior to application of the AXC. During the period of AXC, CSF was allowed to drain freely in an attempt to have cerebrospinal fluid pressure (CSFP) no greater than central venous pressure (CVP). Dogs in Group N were hyperventilated (Paco 2 28-32 mm Hg) and received 2 g/kg of mannitol prior to AXC and then 1 g.kg -1 .hr -1 during clamping. Systemic hemodynamics, CSFP, and arterial blood gases were measured at 1) baseline, 2) 2 min after AXC, 3) 20 min after AXC, 4) 5 min after AXC release, and 5) 30 min after resuscitation. With release of the AXC, Paco 2 was not controlled in Group D ; in Group N the minute ventilation was further increased to maintain Paco 2 constant. At precisely 24 h after AXC, the animals were assessed for incidence and severity of paraplegia, using the Tarlov score, by an observer unaware of the experimental protocol. The animals were then killed, and the entire spinal cord was removed for histologic assessment. Multiple sections of the lumbar spinal cord were processed and stained with hematoxylin and eosin, then examined by light microscopy for nonviable neurons in the anterior spinal cord. At baseline, CSFP was significantly higher in Group D before CSF drainage. The CSFP was significantly less in Group D for all subsequent time periods (treatment X time interaction ; P = 0.0001). However, in Group N, CSFP was always less than baseline values in Group D. There was no difference in spinal cord perfusion pressure (SCPP ; distal mean arterial pressure - CSFP) between groups during cross-clamping. Acidosis was significantly worse in Group D with AXC release. The neurologic outcome was the same in both groups ; Tarlov score was 3 in seven animals and 4 in one animal in each group (P = 1.0 ; Mann-Whitney rank-sum test). There was no difference in the ratio of dead : total lumbar anterior spinal cord neurons between groups (0.010 ± 0.016 and 0.015 ± 0.033 for Groups D and N, respectively ; P = 0.720). We conclude that neuroanesthesia adjunct therapy is as effective as CSF drainage for the prevention of paraplegia in this canine model. Such therapy is less invasive than drainage of CSF and may be of use clinically to decrease the incidence and severity of paraplegia after surgery on the descending thoracic aorta.

The present study examined the maturation of nociceptive primary afferents using expression of Fos-like immunoreactivity in the second-order spinal cord neurons as an anatomical and functional marker. Pinch, immersion in hot water, or formalin injection applied to the hindpaw was used as the peripheral noxious stimulus in awake 0-, 1-, 2-, 3-, and 14-day-old rat pups. On the day of birth, all 3 stimuli elicited expression of the Fos protein in dorsal horn cells indicating that nociceptive primary afferents are functional at this age. The expression of the Fos protein was related to the intensity of stimulation since greater injection volumes of formalin or prolonged application of the thermal stimulus increased the number of stained nuclei. The number of stained nuclei was age dependent and older pups exhibited a greater number of stained nuclei. The results of this study are consistent with electrophysiological studies that have demonstrated that the primary nociceptive afferents continue to mature during the rats' postnatal life. Furthermore, the number of Fos immunoreactive neurons in immature rats are age and stimulus-intensity dependent.

Changes in the metabolic activity of embryonic chick spinal cords were examined following alpha-bungarotoxin (alpha-BTX) administration, in order to investigate a potential mechanism by which this toxin arrests motoneurone apoptosis during neurogenesis. Chick embryos were injected i.p. with alpha-BTX and after 25 h the metabolic markers 2-deoxyglucose and cytochrome oxidase were examined in alternate serial sections of the brachial and lumbar spinal cord. Glucose uptake and cytochrome oxidase activity were reduced throughout the spinal cord and pronounced in the lateral motor columns. Iodinated alpha-BTX reaches and binds to neuronal alpha-BTX-sensitive nicotinic cholinoceptors. Binding of alpha-BTX to these neuronal receptors and to those at the neuromuscular junction has now been shown to have a demonstrable effect on neuronal metabolism. The decreased metabolic activity in spinal cord neurones as a result of toxin treatment may have an important role in the prevention of motoneurone apoptosis at a critical developmental phase.

Hemodilution using modified hemoglobin solutions may reduce ischemic central nervous system injury. Purified diaspirin cross-linked hemoglobin (DCLHb) is a cell-free hemoglobin that is intramolecularly cross-linked between the two alpha subunits, resulting in enhanced oxygen offloading to tissues and increased half-life. In the present experiments, we evaluated the ability of DCLHb to reduce neurological damage in two rabbit stroke models. In a reversible spinal cord ischemia model, ischemia of the caudal lumbar spinal cord was produced by temporary occlusion of the abdominal aorta. In an irreversible model of cerebral ischemia, plastic microspheres (50 microns) were injected into the internal carotid artery and lodged in the cerebral microvasculature. DCLHb was administered 5 minutes after initiation of ischemia as either a 10-mL/kg infusion, 10-mL/kg exchange transfusion, or a 20-mL/kg infusion. Control animals received human serum albumin that was oncotically matched to the DCLHb. In the spinal cord model, DCLHb significantly increased the duration of ischemia required to produce permanent paralysis from 27.33 +/- 8.71 minutes (mean +/- SD) in controls to 42.59 +/- 10.10 minutes in the 10-mL/kg exchange transfusion group and to 40.82 +/- 18.16 minutes in the 20-mL/kg infusion condition (P < .05). DCLHb did not significantly reduce neurological damage in the microsphere embolization model. These data suggest that cross-linked hemoglobin reduces neurological damage after reversible central nervous system ischemia and that this is not attributable to hemodilution or hypervolemia only.

Evoked spinal cord potentials (SCPs) were recorded from the posterior epidural space (PES) at the cervical and lumbrosacral enlargements in response to electrical stimulation of the brachial plexus at Erb's point in 17 chronic pain patients. Erb's point stimulation produced slow positive potentials (heterosegmental slow positive potentials, HSPs) in the PES at the lumbrosacral enlargement in all 13 subjects without spinal cord lesions but not in 4 subjects with spinal cord lesions. The HSP1 with a central peak latency of 21 +/- 2 ms (mean +/- SE) was recorded at the stimulus intensity up to two to three times the threshold strength (T) of the initially positive spike (P1) of the segmental SCP, which was simultaneously recorded from the PES at the cervical enlargement. At the stimulus intensity of more than 3T, another slow positive potential (HSP2) with central peak latency of 71 +/- 6 ms was recorded. These slow positive potentials (HSP1 and HSP2) might be produced by a feedback loop via supraspinal structures, presumably primary afferent depolarizations, in comparison to the HSPs of our previous studies in the rat. Slow negative potentials were sometimes noted before (5 of 13) and/or after (2 of 13) the HSP1. These slow negative potentials probably reflect the activities of dorsal horn neurons producing the HSP1 and HSP2, respectively, also elicited by a feedback loop via supraspinal structures.

Background Epidural opioids must redistribute to the spinal cord to produce analgesia. It has been suggested that opioids may reach the spinal cord via the radicular arterial blood supply. This study was designed to test this hypothesis. Methods Microdialysis probes were placed in the lumbar and thoracic spinal cord of six anesthetized pigs and a ligature placed around the thoracic aorta. At t = 0, fentanyl (300 micrograms) was injected into the epidural space and the spinal cord and plasma were sampled over 90 min to determine resultant spinal cord dialysate fentanyl concentrations. At t = 90, the thoracic aorta was occluded to eliminate distal radicular artery blood flow and fentanyl was again injected into the epidural space. The spinal cord was sampled for an additional 90 min to determine spinal cord dialysate fentanyl concentrations. At t = 90 min the aortic cross-clamp was released and spinal cord samples were collected for another 30 min. Results There was no difference in lumbar spinal cord dialysate concentrations of fentanyl between the control and cross-clamp conditions over the first 20 min after epidural administration. Between 20 and 90 min, lumbar spinal cord fentanyl concentrations were significantly greater during aortic cross-clamp. Fentanyl was not detected in the thoracic cord. Conclusions Radicular artery blood supply does not redistribute fentanyl from the epidural space to the spinal cord in this model. However, radicular artery blood flow does play a role in clearing fentanyl from the spinal cord.

Behavioral and electrophysiological studies have shown that a noxious stimulus applied to one part of the body can reduce the response to a subsequent noxious stimulus elsewhere on the body. This phenomenon is referred to as diffuse noxious inhibitory controls (DNIC). In the present study we used immunocytochemical labeling for the Fos protein product of the c-fos proto-oncogene to determine the location of lumbar spinal nociresponsive neurons that are inhibited by a spatially remote noxious stimulus. Repetitive hindpaw pinch evoked pronounced Fos-like immunoreactivity in the superficial and deep laminae of the lumbar spinal cord. Placing the tail in 50 degrees C water before each hindpaw pinch significantly reduced Fos-like immunoreactivity in these regions. These data demonstrate that nociresponsive neurons in both the superficial and deep laminae of the spinal cord are sensitive to inhibition by a spatially remote noxious conditioning stimulus.

Effects of electrical stimulation of the cervical vagus nerve were determined in cervical or lumbar spinal neurons in 27 rats anesthetized with pentobarbital. Ipsilateral cervical vagus stimulation (ICVS) increased activity of 44 neurons in the C1 segment. At the same stimulation parameters, contralateral cervical vagus stimulation (CCVS) either increased, decreased or did not affect activity of C1 neurons that were excited by ICVS. For C1 cells excited by both ICVS and CCVS, the mean latency for activation was significantly longer for CCVS than for ICVS, and ICVS produced a greater degree of excitation than CCVS. In segments C2-C6, 16 of 18 neurons were excited by ICVS and 2 were inhibited. However, CCVS did not excite the C2-C6 neurons but either inhibited or did not affect activity. In 6 cervical cells, a CCVS conditioning stimulus reduced the level of excitation by ICVS (test stimulus). Transection of the C2 or C3 dorsal roots did not significantly affect the excitatory vagal input to C1 cells. Excitatory somatic receptive fields were classified for 60 cervical spinal cells that responded to vagal stimulation. Most (87%) cells were excited by noxious pinch; 29 were wide dynamic range (WDR) cells and 21 were high threshold cells. In contrast to upper cervical neurons, spinothalamic tract (STT) and spinal cells in lumbar segments were not excited by ICVS or CCVS at the stimulation parameters used in this study, but were primarily inhibited by vagal stimulation. Results of this study showed that a group of cells in upper cervical segments were excited by vagal afferents. This excitatory vagal input reaches the C1 segment primarily via an ipsilateral, supraspinal route.

Spinal cord stimulation has been advocated as an alternative to motor cortex stimulation for motor tract activation. To test this theory, evoked responses were recorded from lumbar spinal cord (L2; n = 14), spinal roots (L4-L7; n = 112), peripheral nerves (sciatics; n = 28), and hind limb muscles (n

Delayed deterioration of neurological function after central nervous system ischemia is a well-documented clinical problem. The purpose of our study was to elucidate the role of spinal cord blood flow and spinal cord-blood barrier integrity in the evolution of delayed neurological deterioration after transient spinal cord ischemia in rabbits. Anesthetized rabbits were subjected to lumbar spinal cord ischemia (25 minutes) and variable periods of reperfusion (30 minutes to 48 hours after ischemia). Regional spinal cord blood flow was monitored by carbon-14-labeled iodoantipyrine autoradiography; vascular permeability was assessed by quantitative microhistofluorescence of Evans blue-albumin in frozen sections of spinal cord. Hindlimb motor function was assessed by standard scoring system and tissue edema by wet/dry weight method. Hindlimb motor function indicated complete paralysis during ischemia and partial gradual recovery upon reperfusion (up to 8 hours), followed by progressive deterioration to severe deficits over 48 hours. Severe vascular permeability disruption was noticed early (30 minutes) after reperfusion, but almost complete recovery reestablished at 8 hours was followed by a secondary progressive increase in vascular permeability. Blood flow was reduced by 20-30% (p less than 0.01) 4 hours after ischemia in the gray matter, but hyperemia (200-300%, p less than 0.01) was observed 12-24 hours after ischemia. Spinal cord water content increased by 5.7% (p less than 0.05) 24 hours after ischemia. This study demonstrates that delayed neurological and motor deterioration after spinal cord ischemia is associated with severe progressive breakdown of spinal cord-blood barrier integrity that develops late (hours) after the injury. Our data suggest that no ischemic insult in early or late reperfusion is associated with delayed motor deterioration.

Objective The objective was to report on the possibility of allergic reaction to the components of a spinal cord stimulator. Design We describe a severe allergic reaction after the insertion of a spinal cord stimulator in a patient with complex regional pain syndrome type 1. Setting The patient was being followed in an office-based pain management practice. Patient The patient is a 41-year-old woman with complex regional pain syndrome type 1, posttrauma. Intervention Insertion of a cervical and lumbar spinal cord stimulator. Outcome measures The outcome measures were a numerical scale of pain intensity and the ability to perform the activities of daily living. Results Adequate pain control complicated by allergic reaction. Conclusions There exists a possibility that a patient may experience an allergic reaction to spinal cord stimulator components. Recognition of such contact sensitivity is important for physicians implanting such devices. Patients may be misdiagnosed as having infections, which can delay appropriate management; definitive diagnosis can be confirmed with a patch test. Treatment consists of removal of such devices.