Down-regulation of mu opioid receptor expression within distinct subpopulations of dorsal root ganglion neurons in a murine model of bone cancer pain.

Although micro opioid receptor (MOR) agonists are used for treatment of most types of pain, a recent study has suggested that the sensitivity of bone cancer pain to systemic morphine was lower than that of inflammatory pain. However, the reasons for this have remained unclear. In this study, MOR expression and the analgesic effects of morphine in a bone cancer model were compared with those in an inflammatory pain model. A bone cancer pain model and an inflammatory pain model were made by implantation of sarcoma cells into the intramedullary space of the femur and hind-paw injection of complete Freund's adjuvant (CFA), respectively. In a behavioral study, sarcoma-implanted mice showed flinching behavior of magnitude comparable to that induced by CFA injection. The flinching behavior of sarcoma-implanted mice was less sensitive to intrathecal morphine than that of CFA-injected mice. Western blot analysis showed that MOR expression in the dorsal root ganglion (DRG) ipsilateral to sarcoma implantation was significantly reduced, while that in the DRG ipsilateral to CFA injection was increased. In sarcoma-implanted mice, the percentage of MOR-positive DRG neuronal profiles was lower than that in control mice (30.3% vs. 45.2%). In particular, MOR expression was reduced among calcitonin gene-related peptide- and transient receptor potential vanilloid subfamily 1-positive DRG neuronal profiles, which are considered to be involved in the generation of bone cancer pain (from 61.5% to 41.5% and from 72.1% to 48.4%, respectively). These results suggest that down-regulation of MOR in the distinct populations of DRG neurons contributes to the fact that higher doses of morphine are needed to produce analgesia in bone cancer as compared with those used in non-malignant inflammatory situations.

Bone cancer increases transient receptor potential vanilloid subfamily 1 expression within distinct subpopulations of dorsal root ganglion neurons.

Bone cancer pain has a strong impact on the quality of life of patients but is difficult to treat. Therefore, the mechanisms of bone cancer pain require elucidation for the purpose of development of new therapeutics. A recent study showed that activation of transient receptor potential vanilloid subfamily 1 (TRPV1) was involved in bone cancer pain. In this study, we re-evaluated the analgesic effects of pharmacological blockade of TRPV1 using the potent TRPV1 antagonist 5-iodoresiniferatoxin (I-RTX) and examined whether bone cancer can change TRPV1 expression and distribution in the primary sensory neurons in a mouse model of bone cancer pain. Implantation of osteosarcoma into the femur induced ongoing and movement-evoked bone cancer-related pain behaviors. These behaviors were significantly reduced by i.p. administration of I-RTX, compared with vehicle. Western blot and reverse transcription-polymerase chain reaction (RT-PCR) analyses revealed that TRPV1 level was significantly increased in dorsal root ganglions (DRGs) ipsilateral to sarcoma implantation. Immunohistochemical analysis showed that implantation of osteosarcoma induced not only an increase in the percentage of TRPV1-positive neurons among DRG neurons (24.3+/-1.3% in sham mice and 31.2+/-1.3% in mice with osteosarcoma implantation, P<0.05) but also an overall shift in the distribution of area of profiles to the right. Colocalization study showed that the percentages of colocalization of TRPV1 with neurofilament 200 kD (NF200) and calcitonin gene-related peptide (CGRP) but not isolectin B4 (IB4) among DRG neurons in mice with osteosarcoma implantation were increased compared with those in sham mice (from 0.8+/-0.1% to 2.1+/-0.3% for TRPV1 and NF200 and from 21.1+/-1.3% to 26.5+/-0.2% for TRPV1 and CGRP). In conclusion, TRPV1 activation plays a critical role in the generation of bone cancer pain, and bone cancer increases TRPV1 expression within distinct subpopulation of DRG neurons. These findings may lead to novel strategies for the treatment of bone cancer pain.

Endothelin-1 enhances capsaicin-evoked intracellular Ca2+ response via activation of endothelin a receptor in a protein kinase Cepsilon-dependent manner in dorsal root ganglion neurons.

Increasing evidence indicates that endothelin-1 has a role for peripheral nociceptive signaling in animals and humans. However, the mechanisms of the nociceptive effects of endothelin-1 have not been fully understood. The current study investigated the effects of endothelin-1 on the capsaicin-evoked intracellular Ca2+ response of cultured adult mice dorsal root ganglion neurons. Dorsal root ganglia were harvested from adult male C57B6N mice and were cultured. With a digital image analysis system, we detected the [Ca2+]i image of cultured dorsal root ganglion cells after loading with Fura-2 acetoxymethyl. In addition, co-localization of protein kinase Cepsilon with transient receptor potential V1 and the translocation of protein kinase Cepsilon were investigated using immunohistochemical methods. Endothelin-1 (10 nM) enhanced an increase in [Ca2+]i by capsaicin (10 nM) from 87.6+/-11.6 nM to 414.8+/-62.3 nM (71 of 156 neurons). The inhibition of endothelin A receptor (BQ-123) significantly suppressed the enhancing effect of endothelin-1. In addition, a nonselective protein kinase C inhibitor (bisindolylmaleimide I) significantly suppressed the enhancing effect of endothelin-1. A myristoyl-tagged membrane-permeant-protein kinase Cepsilon V1-2 inhibitory peptide also significantly suppressed the enhancing effect of endothelin-1. In the immunocytochemical study, protein kinase Cepsilon immunoreactivity was found in most of transient receptor potential V1-positive neurons. After endothelin-1 application, protein kinase Cepsilon immunoreactivity was observed to be translocated from the cytosol to the cell membrane in transient receptor potential V1-positive neurons. Our results indicate that endothelin-1 enhances the response of dorsal root ganglion neurons to capsaicin in a protein kinase Cepsilon-dependent manner. Our findings may lead to a new strategy to treat pain associated with endothelin-1.

Immunohistochemical analysis of acid-sensing ion channel 2 expression in rat dorsal root ganglion and effects of axotomy.

Several studies have suggested that acid-sensing ion channel 2 (ASIC2) plays a role in mechanoperception and acid sensing in the peripheral nervous system. We examined the expression and distribution of ASIC2 in the rat dorsal root ganglion, the co-localization of ASIC2 with tropomyosin-related kinase (trk) receptors, and the effects of axotomy on ASIC2 expression. ASIC2 immunoreactivity was observed in both neurons and satellite cells. ASIC2-positive neurons accounted for 16.5 +/- 2.4% of the total neurons in normal dorsal root ganglion. Most ASIC2-positive neurons were medium-to-large neurons and were labeled with neurofilament 200 kD (NF200). Within these neurons, ASIC2 was not evenly distributed throughout the cytoplasm, but rather was accumulated prominently in the cytoplasm adjacent to the axon hillock and axonal process. We next examined the co-localization of ASIC2 with trk receptors. trkA was expressed in few ASIC2-positive neurons, and trkB and trkC were observed in 85.2% and 53.4% of ASIC2-positive neurons, respectively, while only 6.9% of ASIC2-positive neurons were co-localized with trkC alone. Peripheral axotomy markedly reduced ASIC2 expression in the axotomized dorsal root ganglion neurons. On the other hand, intense ASIC2 staining was observed in satellite cells. These results show that ASIC2 is expressed in the distinct neurochemical population of sensory neurons as well as satellite cells, and that peripheral axotomy induced marked reductions in ASIC2 in neurons.

Central venous catheter migration to the popliteal artery during total cavopulmonary shunt.

Central venous catheter migration to the arterial system occurred due to the surgical procedure during total cavopulmonary shunt. The catheter seems to have crept into the portion of the vena cava that had been designated for clamping, because of the position for the surgery and the anatomic characteristics of the patient. The catheter migrated in the right popliteal artery and was removed immediately; however, it could enter the pulmonary arterial system and the other systemic arterial branches.

Acute pulmonary edema associated with transfusion of packed red blood cells.

We experienced a patient who suffered noncardiogenic acute pulmonary edema after transfusion of packed red blood cells which contained antigranulocyte antibodies. The data suggested that complement activation and the release of polymorphonuclear protease were involved in the pathogenesis of the complication in the present patient. Furthermore, blood coagulative system was also activated after the transfusion. The underlying mechanisms of the complication are discussed.

Involvement of capsaicin-sensitive fibers in spinal NMDA-induced glutamate release.

The activation of spinal NMDA receptors can evoke glutamate release through the production of nitric oxide (NO) within the spinal cord, resulting in pain-relating behavior. In this study, we investigated the involvement of capsaicin-sensitive primary afferents in this phenomenon using in vivo intrathecal microdialysis. Intrathecal NMDA perfusion evoked increases in the concentrations of NO metabolises and glutamate and in pain-related behavior in both neonatal capsaicin and vehicle-treated rats. Although the degrees of increase in NO metabolises in capsaicin- and vehicle-treated rats were not significantly different, capsaicin-treated rats showed significantly smaller increases in glutamate concentration and pain-related behavior than did vehicle-treated rats. Our results showed that glutamate release from capsaicin-sensitive primary afferent terminals is involved in spinal NMDA-induced pain.

Roles of monoaminergic, glycinergic and GABAergic inhibitory systems in the spinal cord in rats with peripheral mononeuropathy.

The current study was designed to determine if the monoaminergic descending inhibitory system and the glycinergic and GABAergic inhibitory systems were activated in the spinal cord in the presence of peripheral mononeuropathy produced by loose ligatures around the common sciatic nerve. The time course of withdrawal latencies to thermal stimuli were assayed in lesioned and sham-operated rats. The levels of monoamines (serotonin; 5-HT, noradrenaline, and dopamine), glycine and gamma-aminobutyric acid (GABA) in the dorsal half of the spinal cord were measured using HPLC with electrochemical detection. Furthermore, on day 7 after nerve ligation, intrathecal methysergide, yohimbine, strychnine or bicuculline was administered in order to investigate the roles of these inhibitory neuromodulators in this pathological pain state. The levels of 5-HT and noradrenaline significantly increased in both ipsi- and contralateral sides of the dorsal half of the lumbar spinal cord in the lesioned, but not sham-operated animals. The levels of glycine and GABA in the ipsilateral dorsal half of the spinal cord increased significantly and were significantly higher than in the contralateral side. Intrathecal antagonists of 5-HT, noradrenaline, glycine and GABA produced enhancement of the magnitude of hyperalgesia on the lesioned hindpaw. We also examined the effects of four daily single treatments with intrathecal MK-801 beginning 15 min prior to nerve ligation on the development of thermal hyperalgesia and on the contents of the neuromodulators in the ligation model. MK-801 treatment effectively abolished the increases in 5-HT, noradrenaline, glycine and GABA levels as well as preventing the development of hyperalgesia. The results of the present study suggest that the pathological pain state activates or increases the activity of these inhibitory systems.

Direct evidence for the role of nitric oxide on the glutamate-induced neuronal death in cultured cortical neurons.

It has been reported that glutamate-induced neurotoxicity is related to an increase in nitric oxide (NO) concentration. An NO-sensitive electrode has been developed to measure NO concentration directly. Using this electrode, we examined NO concentration and neuronal survival after glutamate application in rat cultured cortical neurons. We also examined the effects of NMDA receptor antagonists, MK-801 and ketamine, and the NO synthetase inhibitor, L-NMMA on NO production and neuronal death. After 7 days in culture, application of glutamate (1 mM) or L-arginine (0.3 mM) to the cultured medium increased NO concentration, and decreased the number of anti-microtubule-associated protein 2 positive neurons. Both pretreatment with MK-801 (300 microns) and ketamine (300 microns) prevented glutamate-, but not L-arginine-induced increase in NO concentration and neuronal death. L-NMMA prevented both glutamate- and L-arginine-induced NO production and neuronal death. The nitric oxide donor, S-nitroso-N-acetyl-D,L-penicillamine (SNAP) also caused neuronal death, and MK-801, ketamine and L-NMMA did not prevent SNAP-induced toxicity. We have demonstrated excitatory amino acid-induced changes of NO concentration and the parallel relationship between changes of NO concentration and neuronal death. In conclusion, an increase in NO concentration does induce neuronal death, and the inhibition of the production of NO prevents glutamate-induced neuronal death.

Formalin-induced nociception activates a monoaminergic descending inhibitory system.

Neural plasticity of afferent pain pathways that is induced by prolonged or repeated noxious stimuli may contribute to activate intrinsic inhibitory mechanisms in CNS. In order to clarify the role of the monoaminergic descending inhibitory system in acute nociception and inflammatory pain, we examined if this inhibitory system would modulate the tonic response to formalin-induced nociception. Yohimbine, alpha2 adrenergic antagonist, or methysergide, serotonin antagonist was administered intrathecally before or after subcutaneous 2% formalin injection into the plantar of the hind paw in rats. In another series of the experiment, the tissue of the spinal dorsal half of the untreated rats and post-formalin-treated rats were sampled and analyses of monoamine levels were carried out by HPLC. The subcutaneous formalin evoked biphasic flinching behavior of the injected paw. Intrathecal pretreatment with yohimbine and methysergide produced a significantly greater increase in the number of flinches than in the control in phase 1, intermediate period and phase 2. Posttreatment with yohimbine and methysergide showed a significantly greater increase in the number of flinches in phase 2. Furthermore, formalin injection induced significant increases in noradrenaline, MHPG, serotonin (5-hydroxytryptamine; 5-HT) and 5-HIAA concentrations in both the ipsi- and contralateral dorsal halves. These results suggest that the pain state produced by formalin-induced chemical and/or inflammatory nociception is under the modulation of the monoaminergic (noradrenergic and serotonergic) descending inhibitory system.

Formalin-induced release of excitatory amino acids in the skin of the rat hindpaw.

Application of glutamate to skin evokes pain-related behaviors [S.M. Carlton, G.L. Hargett, R.E. Coggeshall, Localization and activation of glutamate receptors in unmyelinated axons of rat glabrous skin, Neurosci. Lett., 197 (1995) 25-28; D.L. Jackson, C.B. Graff, J.D. Richardson, K.M. Hargreaves, Glutamate participates in the peripheral modulation of thermal hyperalgesia in rats, Eur. J. Pharmacol., 284 (1995) 321-325.] and peripherally-administered glutamate antagonists can prevent the nociception produced by inflammation [E.M. Davidson, R.E. Coggeshall, S.M. Carlton, Peripheral NMDA and non-NMDA glutamate receptors contribute to nociceptive behaviors in the rat formalin test, NeuroReport, 8 (1997) 941-946; Jackson et al., 1995.] In this study, the concentrations of glutamate and aspartate in the plantar of the rat hindpaws were measured before and after the subcutaneous administration of formalin. Increases in glutamate and aspartate concentrations were observed on the ipsilateral side, but not on the contralateral side, to the injection. This shows that nociception and inflammation caused by formalin injection induces the release of peripheral glutamate and aspartate, which would contribute to nociception and inflammatory pain.

Delta receptor involvement in morphine suppression of noxiously evoked activity of spinal WDR neurons in cats.

Morphine has been considered to be primarily a mu opiate receptor agonist. The present study was designed to determine if opiate receptor subtypes in addition to mu contribute to morphine analgesia at the level of the spinal cord. Extracellular activity of single wide dynamic range (WDR) neurons in the feline lumbar spinal cord were studied. Intrathecal administration of DAGO (selective mu agonist) or DPDPE (selective delta agonist) suppressed the noxiously (51 degrees C radiant heat) evoked activity of WDR neurons. Pretreatment with spinal beta-FNA (selective mu antagonist) antagonized the suppressive effects of spinal DAGO, but not that of DPDPE. Two doses of spinal morphine (200 and 400 micrograms) suppressed the noxiously evoked activity of WDR neurons confirming our previous report. Following beta-FNA pretreatment, the suppressive effects of morphine were reduced, however, when ICI 174,864 (selective delta antagonist) was co-administered with morphine on the spinal cord of the animals pretreated by beta-FNA, there was an even greater reduction in the neuronal suppression by morphine. Intravenous ICI 174,864 also reversed the suppressive effects of morphine in beta-FNA pretreated animals. beta-FNA antagonism of spinal morphine is evidence of the well-known mu receptor-mediating antinociception. However, antagonism by ICI 174,864 of morphine suppression in beta-FNA-pretreated animals demonstrates that morphine is capable of suppressing noxiously evoked activity of WDR neurons as a result of an interaction with delta receptors in addition to mu receptors at the level of spinal cord.

Peripheral nitric oxide in carrageenan-induced inflammation.

Recent studies have suggested that nitric oxide (NO) peripherally produced by different nitric oxide synthase (NOS) isoforms contributes to edema formation and development of hyperalgesia. The present study was designed to examine the effects of NOS isoforms on NO release in carrageenan-induced inflammation at various time points. A microdialysis probe was implanted subcutaneously into the glabrous skin of hindpaws of Sprague-Dawley rats under pentobarbital anesthesia. After sample collection to obtain the basal level of the total amount of nitrite and nitrate (NO2-/NO3-), modified Ringer solution, a non-selective NOS inhibitor, NG monomethyl-L-arginine acetate (L-NMMA), or an iNOS inhibitor, aminoguanidine hemisulfate (AG) was perfused through the microdialysis probe. 2 mg of carrageenan was injected into the plantar surface of the probe-implanted hindpaw. Carrageenan was also injected in rats that had undergone sciatic nerve sectioning. Carrageenan significantly increased the dialysate concentrations of NO2-/NO3- for more than 8 h. L-NMMA suppressed the carrageenan-induced increase in NO2-/NO3- concentration. Although AG did not suppress the increase in NO2-/NO3- for the first 2 h after carrageenan injection, significant suppression of the increase in NO2-/NO3- was observed from 2.5 h after carrageenan injection. In the rats in which the sciatic nerves had been denervated, the increases in concentrations of NO2-/NO3- were completely suppressed up to 3 h and partially suppressed 4.5-8 h after carrageenan injection. The results of the current study show that carrageenan induces peripheral release of NO, the production of which is mediated by nNOS in the early phase and by both nNOS and iNOS in the late phase of carrageenan-induced inflammation.

Self-guided robotic camera control for laparoscopic surgery compared with human camera control.

BACKGROUND: In laparoscopic surgery, the surgeon no longer has direct visual control of the operation area, and a camera assistant who maneuvers the laparoscope is essential. Problems of cooperation between the two naturally arise, and a robotic assistant that automatically controls the laparoscope can offer a highly desirable alternative to this situation. METHODS: A self-guided robotic camera control system (SGRCCS) based upon a color tracking method has been developed and its use evaluated in 20 cases of laparoscopic cholecystectomy and compared with using human camera control. RESULTS: In 83% of the patients the procedures were successfully completed with the SGRCCS. Set-up time for the robot averaged 21 minutes; and the surgical time with and without the robot averaged 54 and 60 minutes, respectively. Using the robot instead of a human camera assistant significantly reduced both the frequency of the camera correction, 2.2 per hour compared with 15.3 per hour, and frequency of the lens cleaning, 1.0 per hour compared with 6.8 per hour. Subjective assessment by the surgeon revealed that the robot performed better than the human assistant in 71 % of the cases. CONCLUSIONS: In laparoscopic surgery, the SGRCCS offered optimal camera guidance and helped to maintain the surgeon's concentration during the operation.

Evaluation of cervical cord function in cervical spondylotic myelopathy and/or radiculopathy using both segmental conductive spinal-evoked potentials (SEP).

To evaluate the function of the cervical cord and to diagnose the level and severity of cervical cord lesions in myelopathy, both segmental and conductive spinal evoked potentials (SEP) were measured in 73 patients with cervical spondylotic myelopathy and/or radiculopathy. In normal subjects, segmental SEPs consisted of two waves (R and N waves). Ascending conductive SEPs also consisted of two waves (first and second waves). The function of the cervical cord, including roots, grey matter, and white matter, can be measured by the combined method using both segmental and conductive SEPs, and this allows differentiation among radiculopathy and various types of myelopathy.

Evaluation of cervical cord function using spinal evoked potentials from surface electrode.

Spinal evoked potentials from cervical skin surface (surface spinal evoked potentials) were measured to evaluate spinal cord function as a convenient method that precludes inserting electrodes into the epidural space, and results were compared with those of the former epidural recording method. Surface spinal evoked potentials were obtained from cervical skin surface over the C3, C5, and C7 spinous processes after median nerve stimulation in 18 normal subjects and 37 patients with a cervical lesion. In normal subjects, surface spinal evoked potentials consisted of three negative waves (N1, N2, N3). Abnormal N2 (80%) and abnormal N3 (100%) were observed in cervical myelopathy, and abnormal N2 was observed only in radiculopathy; this allows for differentiation between myelopathy and radiculopathy. Comparing preoperative and postoperative surface spinal evoked potentials, it was seen that improvement of clinical symptoms was proportional to that of surface spinal evoked potentials.

Altered expression of beta-catenin and c-erbB-2 in early gastric cancer.

To investigate the possible relationship between altered expression (loss of membranous staining or nuclear accumulation) of beta-catenin and invasion/metastasis in early gastric cancer (EGC), beta-catenin was detected immunohistochemically in 116 cases of EGC, including 86 differentiated and 30 undifferentiated carcinomas. In parallel, immunohistochemical expression of c-erbB-2 was analyzed in all EGC cases. Regardless of histological type, altered expression of beta-catenin was found in 47% of mucosal carcinomas and 89% of carcinomas with submucosal invasion (p<0.001). Of particular interest is that beta-catenin alteration was found in almost all EGCs with lymph node metastasis, even though no significant statistical comparison could be made. These results suggest that molecular changes resulting in abnormal beta-catenin expression participate in the process of submucosal invasion and metastasis. While loss of expression was preferentially observed in undifferentiated EGCs, nuclear accumulation was found exclusively in 24% of differentiated EGCs. c-erbB-2 was overexpressed in only 16% of differentiated EGCs but there was no correlation between this overexpression and invasion or metastasis. However, it is intriguing that 12 out of 14 cases with c-erbB-2 overexpression also showed altered beta-catenin expression, suggesting that both molecules are involved in the development of a certain set of differentiated EGCs.

Sympathetic ganglion blockade for the management of hyperhidrosis.

We present three patients with severe primary hyperhidrosis, refractory to conservative medical treatment, who were successfully managed with sympathetic ganglion blockade with ethanol. We also summarize 10 patients with hyperhidrosis who underwent sympathetic ganglion blockade in the past 2 years. This closed percutaneous method offers the patients considerably less discomfort and less stress with minimal morbidity and has a efficacy similar to that of surgical sympathectomy, which has previously been the only effective and permanent therapy for severe primary hyperhidrosis. It is concluded that chemical sympathectomy is an effective and useful method for treating severe hyperhidrosis which has advantages over surgical sympathectomy.

Vecuronium neuromuscular blockade at the cricothyroid and posterior cricoarytenoid muscles of the larynx and at the adductor pollicis muscle in humans.

STUDY OBJECTIVE: To compare the sensitivity to vecuronium in the cricothyroid (CT) and posterior cricoarytenoid (PCA) muscles of the larynx and the adductor pollicis muscles. DESIGN: Prospective, nonrandomized study. SETTING: Operating room at Sapporo Medical College and Hospital. PATIENTS: 9 ASA status I and II adult patients scheduled for total laryngectomy to resect laryngeal cancer of essentially unilateral involvement. INTERVENTIONS: During surgery, electromyographic (EMG) recording wire electrodes were inserted into the CT and PCA muscles with nitrous oxide, oxygen, and fentanyl anesthesia. The evoked compound EMG responses of the CT and PCA muscles were quantified simultaneously in an identical manner by supramaximally stimulating the superior laryngeal and recurrent nerves at 0.1 Hz and 0.2 millisecond. The ulnar nerve also was stimulated, and the adduction force of the thumb was measured. Vecuronium was infused continuously at a rate of 0.01 mg/kg/min. The evoked responses (percentage of control, means +/- SD) of the adductor pollicis, CT, and PCA muscles were 12.0 +/- 4.2, 22.0 +/- 5.3, and 32.9 +/- 7.8, respectively, 6 minutes after vecuronium administration. MEASUREMENTS AND MAIN RESULTS: Vecuronium produced significantly more intense neuromuscular blockade in the adductor pollicis than in the laryngeal muscles (p < 0.05). In comparing the laryngeal muscles, the depression of EMG at the CT was significantly greater than that at the PCA (p < 0.05). CONCLUSIONS: The laryngeal (CT and PCA) muscles are more resistant to vecuronium than is the adductor pollicis, and the PCA muscle is more resistant to vecuronium than is the CT muscle. This finding suggests that partially paralyzed patients are able to maintain laryngeal patency despite the apparent impaired neuromuscular transmission in the adductor pollicis muscle.

[Analgesic mechanism of ketamine].

The present study was conducted to investigate the mechanism of antinociceptive action of ketamine in rats. Intraperitoneally administered ketamine showed antinociception in the tail flick (TF) and mechanical paw pressure (MPP) tests in a time- and dose-dependent manner. However, intrathecal ketamine, 50-1,000 micrograms, did not exert any antinociceptive effect in TF and MPP tests. In contrast to the antinociceptive actions of intraperitoneal ketamine in TF test in intact rats, ketamine did not produce any effect in spinally transected rats. Further, pretreatment with an intrathecal serotonin antagonist, methysergide, and an intrathecal alpha 2 adrenergic antagonist, yohimbine, abolished the antinociceptive effects of intraperitoneal ketamine in the TF and the MPP tests. The monoamine levels (noradrenaline, MHPG, 5-HT, 5-HIAA) in the spinal cord, which were measured by high performance liquid chromatography, increased after the intraperitoneal administration of ketamine. We conclude that the antinociceptive effects of ketamine involve an activation of the monoaminergic (noradrenergic and serotonergic) descending inhibitory system, and that spinal ketamine does not affect the responses to the acute noxious stimuli.