Effective treatments for neuropathic pain are lacking due to our limited understanding of the mechanisms. The circRNAs are mainly enriched in the central nervous system. However, their function in various physiological and pathological conditions have yet to be determined. Here, we identified circFhit, an exon-intron circRNA expressed in GABAergic neurons, which reduced the inhibitory synaptic transmission in the spinal dorsal horn to mediate spared nerve injury-induced neuropathic pain. Moreover, we found that circFhit decreased the expression of GAD65 and induced hyperexcitation in NK1R⁺ neurons by promoting the expression of its parental gene Fhit in cis. Mechanistically, circFhit was directly bound to the intronic region of Fhit, and formed a circFhit/HNRNPK complex to promote Pol II phosphorylation and H2B monoubiquitination by recruiting CDK9 and RNF40 to the Fhit intron. In summary, we revealed that the exon-intron circFhit contributes to GABAergic neuron-mediated NK1R⁺ neuronal hyperexcitation and neuropathic pain via regulating Fhit in cis.
Rats ; Animals ; Posterior Horn Cells/pathology* ; Spinal Cord Dorsal Horn/metabolism* ; Neuralgia ; Synaptic Transmission

Motor evoked potential(MEP) produced by cortical surface or transcranial stimulation has evolved as a new clinical and experimental tool to monitor the integrity of motor pathways and to map motor cortex. Clinical assessment of motor system using MEP has further advanced with recent development of the magnetic stimulator. The primary concept using MEPs for test of motor pathways was based on the assumption that pyramidal neurons in the motor cortex are activated by electrical stimulation applied on the cerebral cortex and synchronized compound action potentials are conducted mainly along the corticospinal tracts in the spinal cord. However,recent studies indicated that the origins of the Meps in non primates may differ from those previously believed. In order to use MEPs as a clinical or experimental tool, it is essential to clarify the origin of MEPs. Therefore, goals of this study were : (1) to investigate the origin of MEPs, and (2) to design the most reliable but simple method to evoke and monitor MEPs. In a total of fifteen rats, MEPs were produced by cortex to cortex stimulation and were monitored using a pair of epidural electrodes. Using varying stimulus intensities, the amplitudes and latencies of MEPs were statistically analyzed. The latencies and amplitudes of the MEPs in these animals showed surprisingly large standard deviations, which were partially resulted in these animals showed surprisingly large standard deviations, which were partially resulted from convergence of neighboring waves during high stimulation intensities. Wave forms of MEPs were also varied greatly depending on the position of recording electordes. At low stimulus intensities, most consisten MEPs were obtained when the stimulating electrodes were placed on the hard palate and the temporal muscle, not on the motor cortex. This observation indicates that the primary source of MEPs is not the motor cortex in the rat. When the potentials generated by direct stimulation of motor cortex and those generated by reticular nuclei were monitored epidurally in the same preparation using the same electrodes, these potentials generated by different sources actually identical in their latencies and wave forms. However, the threshold stimulus intensities evoking these potentials were quite different in the two metholds. The threshold was much lower to evoke potentails by reticular nuclei stimulation. It suggests that MEPs are geneated by the reticular nuclei or brain structure located in the brain stem. The observation that the motor cortex play no major roles in generating MEPs was confirmed by sequential sections of neural axis from the motor cortex to brain stem in three rats. All these findings suggested that neither direct motor cortex stimulation not transcranial stimulation did evoke MEPs originating from the motor cortex in rat. These stimulating methods activate reticular nuclei by stimulus current spread to the brain stem. Since the reticular formation plays an important role in motor function in rats, MEP originated from reticular nucleus can be an important testing of the motor function in rats. Moreover, transcranial stimulation of the brain is technically easy. This technique producing MEPs originated from reticular nucleus can be useful to monitor the integrity of motor pathways.
Action Potentials ; Animals ; Axis, Cervical Vertebra ; Brain ; Brain Stem ; Cerebral Cortex ; Efferent Pathways ; Electric Stimulation ; Electrodes ; Evoked Potentials, Motor* ; Extrapyramidal Tracts ; Motor Cortex ; Neurons ; Palate, Hard ; Primates ; Pyramidal Tracts ; Rats* ; Reticular Formation ; Spinal Cord ; Temporal Muscle

Epidural injection of 2-4mg morphine for 7-14 days were given to 18 patients with severe chronic pain. All cases except 2 arachnoiditis had considerable amelioration of pain, which commenced within 2-3min, reached a peak in 10-20min, and was effective for 6-72 hours. Using sensory evoked potentials(SEP) examination for pre-and post-morphine epidural infusion in 18 pain patients, we found the morphine inhibited the action of spinal pain receptor system. It is suggested that morphine reached the subarachnoid space and produced its effect by direct action on the specific opiate receptors in the substantia gelatinosa of the posterior horn cell of the spinal cord.
Arachnoid ; Arachnoiditis ; Chronic Pain* ; Humans ; Injections, Epidural ; Morphine* ; Nociceptors ; Posterior Horn Cells ; Receptors, Opioid ; Spinal Cord ; Subarachnoid Space ; Substantia Gelatinosa

Caudal narcotic analgesia was assessed after the injection of 3mg morphine diluted in 30ml(physiologic) saline into the sacral canal in 15 patients after upper abdominal surgery, in 20 patients after lower abdominal surgery under general anesthesia, and in 20 patients after perianal surgery under caudal block. Pain relief was evaluated by the subsequent need for systemic analgesics. All cases had considerable relief from pain and the morphine was effective for 12 or more hours. There were no significant differences between pain relief of the upper abdominal and lower abdominal surgery group, upper abdomianl and perianal surgery group, and lower abdominal and perianal surgery group (p>0.05, p>0.05, p>0.05). It is suggested that the morphine, which was administered into the sacral, cannal, reached the subarachnoid space and produced it's effect by direct action on the specific opiate receptors in the substantia gelatinosa of the posterior horn cell of the spinal cord. Consequently, whether analgesia from epidural narcotics appears to be segmental in distribution or not is still in controveray.
Analgesia ; Analgesics ; Anesthesia, General ; Humans ; Morphine* ; Narcotics ; Pain, Postoperative* ; Posterior Horn Cells ; Receptors, Opioid ; Spinal Cord ; Subarachnoid Space ; Substantia Gelatinosa

Recently, motor evoked potential(MEP) using cortical surface of transcranial stimulation have been used to monitor the integrity of motor pathways and map motor cortex in human and animal. The primary concept using motor evoked potentials(MEPs) for test of motor pathways was based on the assumtion that pyramidal neurons in the motor cortex are activated by electrical stimulation applied on the cerebral cortex and synchronized compound action potentials are conducted mainly along the corticospinal tracts in the spinal cord. However, the origins and the descending pathways of these MEPs in small animals may be different from those of potentials evoked by intracortical microstimulation because of current spread. Our previous study revealed that the origns of the MEPs in rats differed from those previously believed and may be reticular nuclei. To further clarify those results and localize the intraspinal pathways conduction MEPs, consecutive vertical and/or horizontal sections of the spinal cord were performed at T9 cord level in twelve rats. MEPs were recorded at T2/3 and L2/3 before and after each section and sequential alterations of MEPs were observed. In six rats, the stimulation was alternated between the right and left cortex and the lateralities of conduction pathways were compared. All six cases showed no differences of MEPs and pattern of wave abolition after each section between right and left brain stimulation. The alteration of MEPs after each consecutive section was categorized by analyzing latency shift, amplitude change, and disappearance of waves. We divided a cross section of T9 spinal cord into forty-six squares. If one of the categorized changes occurrd after cutting an area, the appropriate score was given for the area since more change of waves meant more significant contribution of the cut area to conduction of MEPs. The score of twelev rats were summed in each forty-six spots and map showing the distribution of MEPs was constructed. The map revealed that MEPs were conducted along the wide area of ventral and lateral funiculus of the spinal cord but mainly along the medial portion of the ventral funiculus of the spinal cord but mainly along the medial portion of the ventral funiculus and ventral portion of the larteral funiculus through which reticulospinal and vestibulospinal tracts pass. No conduction of MEPs along the corticospinal tracts was confirmed. This finding supports the result of our previous study. However, this extrapyramidal MEP conducted along ventral spinal cord in addition to somatosensory evoked potential(SSEP) which is conducted along posterior funiculus can be useful to monitor the integrity of the whole spinal cord. Moreover, the extrapyramidal MEP can be more useful than pyramidal MEP in rats because the reticular formation plays a more important role in motor function and pyramidal tract is located in posterior funiculus.
Action Potentials ; Animals ; Brain ; Cerebral Cortex ; Efferent Pathways ; Electric Stimulation ; Evoked Potentials, Motor* ; Extrapyramidal Tracts ; Humans ; Motor Cortex ; Neurons ; Pyramidal Tracts ; Rats* ; Reticular Formation ; Spinal Cord

OBJECTIVE: The purpose of this study was to evaluate pain-related behaviors after bilateral C2 root resection and change in pain patterns in the suboccipital region in rats.METHODS: Male Sprague-Dawley rats were randomly assigned to three groups (n=25/group); näive, sham, and C2 resection. Three, 7, 10, and 14 days after surgery, cold allodynia was assessed using 20 μL of 99.7% acetone. c-Fos and c-Jun were immunohistochemically stained to evaluate activation of dorsal horn gray matter in C2 segments of the spinal cord 2 hours, 1 day, 7 days, and 14 days after surgery.RESULTS: Three days after surgery, the response to acetone in the sham group was significantly greater than in the näive group, and this significant difference between the näive and sham groups was maintained throughout the experimental period (p < 0.05 at 3, 7, 10, and 14 days). Seven, 10, and 14 days after surgery, the C2 root resection group exhibited a significantly greater response to acetone than the näive group (p < 0.05), and both the sham and C2 resection groups exhibited significantly greater responses to acetone compared with 3 days after surgery. No significant difference in cold allodynia was observed between the sham and C2 root resection groups throughout the experimental period. Two hours after surgery, both the sham and C2 root resection groups exhibited significant increases in c-Fos- and c-Jun-positive neurons compared with the naive group (p=0.0021 and p=0.0358 for the sham group, and p=0.0135 and p=0.014 for the C2 root resection group, respectively). One day after surgery, both the sham and C2 root resection groups exhibited significant decreases in c-Fos -positive neurons compared with two hours after surgery (p=0.0169 and p=0.0123, respectively), and these significant decreases in c-Fos immunoreactivity were maintained in both the sham and C2 root resection groups 7 and 14 days after surgery. The sham and C2 root resection groups presented a tendency toward a decrease in c-Jun-positive neurons 1, 7, and 14 days after surgery, but the decrease did not reach statistical significance.CONCLUSION: We found no significant difference in cold allodynia and the early expression of c-Fos and c-Jun between the sham and C2 resection groups. Our results may support the routine resection of the C2 nerve root for posterior C1–2 fusion, but, further studies are needed.
Acetone ; Animals ; Gray Matter ; Humans ; Hyperalgesia ; Male ; Neuralgia ; Neurons ; Rats ; Rats, Sprague-Dawley ; Spinal Cord ; Spinal Cord Dorsal Horn

Herpes zoster is characterized by multiple grouped erythematous vesicular eruption on the skin with severe pain. Therefore diagnasis of herpes zoster is clinically easy but effective control of its severe pain is not clear yet. Recently, Behar suggested that morphine which was injected into the epidural space reached subarachnoid space and produced potent analgesic effect by direct action on the specific opiate receptors in the substantia gelatinosa of the posterior horn cell of the spinal cord. On the basis suggestion, we have attempted to treat the severe pain of herpes zoster which was developed on right shoulder, right upper anterior and posterior chest, and posterior auricular area. At the initial block, we inserted the catheter into the 6th cervical intervertebral space and the tip of the catheter was placed in epidural space to 3cm cephalad. Then 3mg of morphine mixed with 10ml of normal saline was adminietered and also right stellate ganglion block with 0. 25% bupivacaine 7ml was combined, therefore effective pain relief was obtained for 11 hours. At second block, additional dose of 2mg of morphine mixed with 10ml of normal saline was administered through the inserted epidural catheter at the initial block without stellate ganglion block, and effective pain relief was obtained for 15 hours. After then, whenever the pain was developed, additional dose of 2mg of morphine was administered as the same procedure of the second block, and the duration of pain relief was extended gradually by the each procedure (for 24 hours after third block and for 49 hours after fourth block). At 5 days after initial block, severe pain was completely subsided. Side effects such as postural hypotension, motor weakness and dizziness was not developed except slightly motor weakness after initial block. This result showed that small dose of morphine injection into the epidural space is effective control of severe pain of herpes zoster.
Bupivacaine ; Catheters ; Dizziness ; Epidural Space ; Herpes Zoster* ; Hypotension, Orthostatic ; Morphine* ; Posterior Horn Cells ; Receptors, Opioid ; Shoulder ; Skin ; Spinal Cord ; Stellate Ganglion ; Subarachnoid Space ; Substantia Gelatinosa ; Thorax

The autonomic nervous system (ANS) integrates the function of the internal organs for the homeostasis against various external environmental changes. The efferent components of the ANS are regulated by sensory signals arising from the viscera as well as non-visceral organs. The central neural networks that integrate these sensory signals and modify visceral motor output are complex, and synaptic reflexes formed in the brainstem and spinal cord integrate behavioral responses and visceral responses through the central neural networks. A detailed understanding of the neural network presented above may explain the role of the vestibular system on the homeostasis more extensively.
Autonomic Nervous System ; Brain Stem ; Homeostasis ; Physiology ; Reflex ; Solitary Nucleus ; Spinal Cord ; Spinal Cord Lateral Horn ; Viscera

The caudal subnucleus of the spinal trigeminal nucleus (medullary dorsal horn; MDH) receives direct inputs from small diameter primary afferent fibers that predominantly transmit nociceptive information in the orofacial region. Recent studies indicate that reactive oxygen species (ROS) is involved in persistent pain, primarily through spinal mechanisms. In this study, we aimed to investigate the role of xanthine/xanthine oxidase (X/XO) system, a known generator of superoxide anion (O₂(·−)), on membrane excitability in the rat MDH neurons. For this, we used patch clamp recording and confocal imaging. An application of X/XO (300 µM/30 mU) induced membrane depolarization and inward currents. When slices were pretreated with ROS scavengers, such as phenyl N-tert-butylnitrone (PBN), superoxide dismutase (SOD), and catalase, X/XO-induced responses decreased. Fluorescence intensity in the DCF-DA and DHE-loaded MDH cells increased on the application of X/XO. An anion channel blocker, 4,4-diisothiocyanatostilbene-2,2-disulfonic acid (DIDS), significantly decreased X/XO-induced depolarization. X/XO elicited an inward current associated with a linear current-voltage relationship that reversed near −40 mV. X/XO-induced depolarization reduced in the presence of La³⁺, a nonselective cation channel (NSCC) blocker, and by lowering the external sodium concentration, indicating that membrane depolarization and inward current are induced by influx of Na⁺ ions. In conclusion, X/XO-induced ROS modulate the membrane excitability of MDH neurons, which was related to the activation of NSCC.
Animals ; Catalase ; Facial Pain ; Fluorescence ; Ions ; Membranes ; Neurons* ; Oxidoreductases ; Posterior Horn Cells* ; Rats* ; Reactive Oxygen Species* ; Sodium ; Spinal Cord Dorsal Horn* ; Superoxide Dismutase ; Superoxides ; Trigeminal Nucleus, Spinal ; Xanthine Oxidase

Reactive oxygen species (ROS) and nitrogen species (RNS) are involved in cellular signaling processes as a cause of oxidative stress. According to recent studies, ROS and RNS are important signaling molecules involved in pain transmission through spinal mechanisms. In this study, a patch clamp recording was used in spinal slices of rats to investigate the action mechanisms of O₂˙⁻ and NO on the excitability of substantia gelatinosa (SG) neuron. The application of xanthine and xanthine oxidase (X/XO) compound, a ROS donor, induced inward currents and increased the frequency of spontaneous excitatory postsynaptic currents (sEPSC) in slice preparation. The application of S-nitroso-N-acetyl-DLpenicillamine (SNAP), a RNS donor, also induced inward currents and increased the frequency of sEPSC. In a single cell preparation, X/XO and SNAP had no effect on the inward currents, revealing the involvement of presynaptic action. X/XO and SNAP induced a membrane depolarization in current clamp conditions which was significantly decreased by the addition of thapsigargin to an external calcium free solution for blocking synaptic transmission. Furthermore, X/XO and SNAP increased the frequency of action potentials evoked by depolarizing current pulses, suggesting the involvement of postsynaptic action. According to these results, it was estblished that elevated ROS and RNS in the spinal cord can sensitize the dorsal horn neurons via pre- and postsynaptic mechanisms. Therefore, ROS and RNS play similar roles in the regulation of the membrane excitability of SG neurons.
Action Potentials ; Animals ; Calcium ; Excitatory Postsynaptic Potentials ; Humans ; Membranes ; Neurons ; Nitric Oxide ; Nitrogen ; Oxidative Stress ; Posterior Horn Cells ; Rats ; Reactive Oxygen Species ; Spinal Cord ; Substantia Gelatinosa ; Superoxides ; Synaptic Transmission ; Thapsigargin ; Tissue Donors ; Xanthine ; Xanthine Oxidase