• The Journal of Korean Physical Therapy
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• v.15 no.4
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• pp.299-311
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• 2003

The purpose of this study is to investigate and analysis effect of TENS with immunohistochemistry methode through changes of substance P in spinal using arthritis model after inducing inflammation. The changes of substance P induced at that time are compared with control which is not induced arthritis by means of counting. The effect of TENS (4Hz, $200{\mu}$, 20minutes) is also tested by observing changes of substance P in spinal dorsal horn after application on knee joint of rats which is arthritis model induced by kaolin and carrageenan. The results of this study were as follows: 1. Substance P immunoreactive positive neurons are increased in dorsal horn after inducting arthritis. 2. In arthritis group, Substance P immunoreactive positive neurons are progressively increased from the first to the third days. 3. Substance P immunoreactive positive neurons after applicating TENS on arthritis group are more decreased than only arthritis-induced group. 4. Substance P immunoreactive positive neurons were significantly decreased on the second days resulting from TENS application from the first to the third days. Therefore, TENS application is decrease Substance P immunoreactive positive neurons in spinal dorsal horn of rats induced arthritis. This decrease is considered as analgesic effect of TENS.

• Animal cells and systems
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• v.4 no.4
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• pp.353-359
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• 2000

Location of the neurons in the lateral reticular nucleus projecting to dorsal horn of the cervical, thoracic, or lumbar spinal cord was investigated in the rat using the technique of retrograde transport of horseradish peroxidase. The projection was bilateral with ipsilateral predominance. Neurons projecting to the cervical spinal cord were located near the medial, dorsal, and lateral perimeter of the magnocellular division of the lateral reticular nucleus, whereas cells projecting to the thoracic and lumbar spinal cord were localized in the medial and dorsal boundaries of the magnocellular division. The labeled neurons were distinctly multipolar in shape and measured approximately 10-15 $\mu m$ in their greatest transverse diameter. A few neurons were also observed in the subtrigeminal nucleus, whereas few cells were in the parbocellular division. These observations provide an anatomical substrate for the functional implication of the lateral reticular nucleus in the regulation of spinal nociceptive transmission and vascular hemodynamics via the descending pathway into the spinal cord.

• The Korean Journal of Pain
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• v.11 no.1
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• pp.23-29
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• 1998

Background: The present study was conducted to investigate effects of microinjection of bicuculline, GABA-A receptor antagonist, into the brain stem nuclei on the dorsal horn neuron responsiveness in rats with an experimental peripheral neuropathy. Methods: An experimental neuropathy was induced by a unilateral ligation of L5~L6 spinal nerves of rats. After 2~3 weeks after the surgery, single-unit recording was made from wide dynamic range (WDR) neurons in the spinal cord dorsal horn. Results: Responses of WDR neurons to both noxious and innocuous mechanical stimuli applied to the somatic receptive fields were enhanced on the nerve injured side. These enhanced responsiveness of WDR neurons were suppressed by microinjection of bicuculline into periaqueductal gray(PAG) or nucleus reticularis gigantocellularis(Gi). A similar suppression was also observed when morphine was microinjected into PAG or Gi. Suppressive action by Gi-bicuculline was reversed by naloxonazine, ${\mu}$-opioid receptor antagonist, microinjected into PAG whereas PAG-bicuculline induced suppression was not affected by naloxonazine injection into Gi. Gi-bicuculline induced suppression were reversed by a transection of dorsolateral funiculus(DLF) of the spinal cord. Conclusions: The results suggest that endogenous opioids, via acting on GABAergic interneurons in PAG and Gi, may be involved in the control of neuropathic pain by activating the descending inhibitory pathways that project to the spinal dorsal horn through DLF to inhibit the responsiveness of WDR neurons.

• Proceedings of the Korean Society of Applied Pharmacology
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• 1996.04a
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• pp.175-175
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• 1996

The spinal dorsal horn is the area where primary afferent fibers terminate and cutaneous sensory information is Processed. A number of putative neurotransmitter substances, including excitatory and inhibitory amino acids and peptides, are present in this region and sites and cellular mechanisms of their actions have been a target of numerous studies. In this study, single neurons were acutely isolated and the properties of whole cell current and responses to excitatory and inhibitory neurotransmitters were studied by the patch clamp method. Young rats (7-14 days) were anesthetized with diethyl-ether, and the lumbar spinal cord was excised and cut transversely at a thickness of 30$\mu\textrm{m}$ by Vibroslicer. The treatment of spinal slices with low concentration of proteases (pronase and thermolysin 0.75 mg/$m\ell$) and mechanical dissociation yielded isolated neurons with near intact morphology. Multipolar, ellipsoidal and bipolar, and pyramidal cells were shown. By applying step voltage pulses to neurons held at -70 mV, two types of inward currents and one outward currents observed. The fast activating and inactivating inward current was the Na$\^$+/ current because of its fast kinetics and blocking by 0.5${\mu}$M TTX, a specific blocker of Na$\^$+/ channel. The second type of inward currents were sustained. Based on their kinetics and current-voltage relations, it was likely that the second type of inward current was the voltage-dependent Ca$\^$2+/ current. In the presence of TTX, the steady-state currents mainly represented outward K$\^$+/ current which looked like the delayed rectifier K$\^$+/ current. In addition, the membrane currents produced by agonist of excitatory amino acid (EAA) receptor and the endogenous transmitter candidate L-glutamate were recorded in isolated whole-cell voltage clamped neurons as well as responses to inhibitory amino acids (${\gamma}$-amino butyric acid, glycine). Drugs were applied by a method that allows complete exchange of the solution within 1 sec; an infinite number of solutions can be applied to a single cell.

• The Korean Journal of Physiology and Pharmacology
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• v.12 no.5
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• pp.253-258
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• 2008

Somatostatin (SOM) is a widely distributed peptide in the central nervous system and exerts a variety of hormonal and neural actions. Although SOM is assumed to play an important role in spinal nociceptive processing, its exact function remains unclear. In fact, earlier pharmacological studies have provided results that support either a facilitatory or inhibitory role for SOM in nociception. In the current study, the effects of SOM were investigated using anesthetized cats. Specifically, the responses of rostrally projecting spinal dorsal horn neurons (RPSDH neurons) to different kinds of noxious stimuli (i.e., heat, mechanical and cold stimuli) and to the $A{\delta}$ -and C-fiber activation of the sciatic nerve were studied. Iontophoretically applied SOM suppressed the responses of RPSDH neurons to noxious heat and mechanical stimuli as well as to C-fiber activation. Conversely, it enhanced these responses to noxious cold stimulus and $A{\delta}$-fiber activation. In addition, SOM suppressed glutamate-evoked activities of RPSDH neurons. The effects of SOM were blocked by the SOM receptor antagonist cyclo-SOM. These findings suggest that SOM has a dual effect on the activities of RPSDH neurons; that is, facilitation and inhibition, depending on the modality of pain signaled through them and its action site.

• The Korean Journal of Physiology and Pharmacology
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• v.1 no.3
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• pp.241-249
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• 1997

The aim of the present study is to examine the brainstem sites where the electrical stimulation produces a suppression of dorsal horn neuron responses of neuropathic rats. An experimental neuropathy was induced by a unilateral ligation of L5-L6 spinal nerves of rats. Ten to 15 days after surgery, the spinal cord was exposed and single-unit recording was made on wide dynamic range (WDR) neurons in the dorsal horn. Neuronal responses to mechanical stimuli applied to somatic receptive fields were examined to see if they were modulated by electrical stimulation of various brainstem sites. Electrical stimulation of periaqueductal gray (PAG), n. raphe magnus (RMg) or n. reticularis gigantocellularis (Gi) significantly suppressed responses of WDR neurons -to both noxious and non-noxious stimuli. Electrical stimulation of other brainstem areas, such as locus coeruleus. (LC) and n. reticularis paragigantocellularis lateralis (LPGi), produced little or no suppression. Microinjection of morphine into PAG, RMg, or Gi also produced a suppression as similar pattern to the case of electrical stimulation, whereas morphine injection into LC or LPGi exerted no effects. The results suggest that PAG, NRM and Gi are the principle brainstem nuclei involved in the descending inhibitory systems responsible for the control of neuropathic pain. These systems are likely activated by endogenous opioids and exert their inhibitory effect by acting on WDR neurons in the spinal cord.

• The Korean Journal of Pain
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• v.13 no.1
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• pp.19-30
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• 2000

Background: Partial nerve injury to a peripheral nerve may induce the development of neuropathic pain which is characterized by symptoms such as spontaneous burning pain, allodynia and hyperalgesia. Though underlying mechanism has not fully understood, sensitization of dorsal horn neurons may contribute to generate such symptoms. Nitric oxide acts as an inter- and intracellular messenger in the nervous system and is produced from L-arginine by nitric oxide synthase (NOS). Evidence is accumulating which indicate that nitric oxide may mediate nociceptive information transmission. Recently, it has been reported that NOS inhibitor suppresses neuropathic pain behavior in an neuropathic pain animal model. This study was conducted to determine whether nitric oxide could be involved in the sensitization of dorsal horn neurons in neuropathic animal model. Methods: Neuropathic animal model was made by tightly ligating the left L5 and L6 spinal nerves and we examined the effects of iontophoretically applied NOS inhibitor (L-NAME) on the dorsal horn neuron's responses to mechanical stimuli within the receptive fields. Results: In normal animals, NOS inhibitor (L-NAME) specifically suppressed the responses to the noxious mechanical stimuli. In neuropathic animals, the dorsal horn neuron's responses to mechanical stimuli were enhanced and NOS inhibitor suppressed the dorsal horn neuron's enhanced responses to non-noxious stimuli as well as those to noxious ones. Conclusions: These results suggest that nitric oxide may mediate nociceptive transmission in normal animal and also mediate sensitization of dorsal horn neurons in neuropathic pain state.

• The Korean Journal of Physiology and Pharmacology
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• v.2 no.1
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• pp.9-19
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• 1998

The present study was primarily carried out to characterize the properties of the spinomesencephalic tract (SMT) neurons that project from the upper cervical spinal segments to the midbrain. It was also investigated whether these neurons received convergent afferent inputs from other sources in addition to cervical inputs. Extracellular single unit recordings were made from neurons antidromically activated by stimulation of midbrain. Recording sites were located in lamina $I{\sim}VIII\;of\;C1{\sim}C3$ segments of spinal cord. Receptive field (RF) and response properties to mechanical stimulation were studied in 71 SMT neurons. Response profiles were classified into six groups: complex (Comp, n=9), wide dynamic range (WDR, n=16), low threshold (LT, n=5), high threshold (HT, n=6), deep/tap (Deep, n=10), and non- responsive (NR, n=25). Distributions of stimulation and recording sites were not significantly different between SMT groups classified upon their locations and/or response profiles. Mean conduction velocity of SMT neurons was $16.7{\pm}1.28\;m/sec$. Conduction velocities of SMTs recorded in superficial dorsal horn (SDH, n=15) were significantly slower than those of SMTs recorded in deep dorsal horn (DDH, n=18), lateral reticulated area (LRA, n=21), and intermediate zone and ventral horn (IZ/VH, n=15). Somatic RFs for SMTs in LRA and IZ/VH were significantly larger than those in SDH and DDH. Five SMT units (4 Comps and 1 HT) had inhibitory somatic RFs. About half (25/46) of SMT units have their RFs over trigeminal dermatome. Excitabilities of 5/12 cells and 9/13 cells were modulated by stimulation of ipsilateral phrenic nerve and vagus nerve, respectively. These results suggest that upper cervical SMT neurons are heterogenous in their function by showing a wide range of variety in location within the spinal gray matter, in response profile, and in convergent afferent input.

• The Korean Journal of Pain
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• v.9 no.2
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• pp.326-335
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• 1996

Background: Though a number of studies have described the distribution of substance P(SP)-like immunoreactivity in the spinal cord, they have been focused on lamina I and II of the dorsal horn and there are little morphological studies on the topographic distribution and ultrastructure of the SP immunoreactive neurons especially in the ventral horn of the spinal cord. this study was conducted to identify distribution pattern of SP immunoreactive neurons and to difine the synaptic organization of their processes in ventral horn of the thoracic cord of the cat by preembbeding immunocytochemical method using SP antiserum. Methods: Five adults cats of either sex were used and deeply anesthetized by intramuscular injection of ketamine. After removal of the spinal cord, samples of thoracic cord were taken and placed in fresh fixative at $4^{\circ}C$ for 2 hours. Transverse sections $50{\mu}m$ thick were processed using the preembbeding immunocytochemical method and incubated consecutively in the specific primary antibody and the 10% normal goat serum, the rabbit anti-substance P antiserum, the biotin-labelled goat anti-rabbit IgG and finally the avidin-biotin-peroxidase complex. The processed tissue sections were throughly washed and stained in the black with 1% uranyl acetate. Section were examined on a electron microscope. Results: 1) SP immunoreactive neurons were observed in the gray matter around central canal. 2) In lamina I and II SP immunoreactivity was observed in both myelinated and unmyelinated nerve fibers, but in ventral horn only in the unmyelinated nerve fibers. 3) SP immunoreactive axon terminals with small round and large dense core vesicles made chemical synapses onto the dendrites of motor neurons in the ventral horn. Conclusion: SP immunoreactive neurons might play an important role in modulation of motor neurons in the ventral horn of the thoracic cord of the cat.

• The Korean Journal of Physiology and Pharmacology
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• v.2 no.2
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• pp.155-163
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• 1998

The purpose of present study was to compare the effects of somatostatin (SOM) and morphine (Mor) on the responses of wide dynamic range (WDR) cells to peripheral noxious stimulation. Single neuronal activity was recorded with a carbon-filament electrode at the lumbosacral enlargement of cat spinal cord. After identifying WDR cells, their responses to peripheral noxious mechanical or thermal stimuli were characterized and the effects of SOM and Mor, applied either iontophoretically or intrathecally, were studied. In most cells SOM and Mor suppressed noxious stimulus-evoked WDR neuronal activity, though a few WDR neurons showed no change or were excited by SOM and Mor. Systemically applied naloxone, a non-specific opioid antagonist, always reversed the Mor induced suppression of neuronal activity evoked by noxious mechanical stimuli, but did not always reverse the suppression of neuronal activity elicited by SOM. The suppressive effect of Mor on thermal stimulus-evoked neuronal activity was partially reversed by naloxone, while that of SOM were not reversed at all. The above results suggest that both Mor and SOM exert an inhibitory effect on thermal and mechanical stimulus-evoked WDR neuronal activity in cat spinal dorsal horn, but the mechanisms are dependent upon the functional populations of dorsal horn nociceptive neurons.