• Journal of Korean Ophthalmic Optics Society
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• 제19권2호
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• pp.271-277
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• 2014

Purpose: The purpose of this study was to investigate the immunoreactivity of calretinin in Brazilian opossum (Monodelphis domestica) retina. Calcium-binding protein calretinin is known to play a key role in calcium-mediated signal transduction. Methods: Experiments have been performed by standard immunocytochemical techniques on retina of the Brazilian opossum. Results: Calretinin-immunoreactivity was exhibited within the horizontal subpopulations, AII amacrine and ganglion cell subpopulations in the Brazilian opossum retina. Especially, all calretinin-immunoreactive AII amacrine cells also expressed parvalbumin. Conclusions: Similar to other mammalian retinas, calretinin-immunoreactivity was also observed within the AII amacrine cells in the Brazilian opossum retina. Thus, calretinin can be a marker of AII amacrine cells in the Brazilian opossum retina.

• Development and Reproduction
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• 제17권4호
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• pp.389-397
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• 2013

FGF9/16/20 signaling pathway specify the developmental fates of notochord, mesenchyme, and neural cells in ascidian embryos. Although a conserved Ras/MEK/Erk/Ets pathway is known to be involved in this signaling, the detailed mechanisms of regulation of FGF signaling pathway have remained largely elusive. In this study, we have isolated Hr-Erf, an ascidian orthologue of vertebrate Erf, to elucidate interactions of transcription factors involved in FGF signaling of the ascidian embryo. The Hr-Erf cDNA encompassed 3110 nucleotides including sequence encoded a predicted polypeptide of 760 amino acids. The polypeptide had the Ets DNA-binding domain in its N-terminal region. In adult animals, Hr-Erf mRNA was predominantly detected in muscle, and at lower levels in ganglion, gills, gonad, hepatopancreas, and stomach by quantitative real-time PCR (QPCR) method. During embryogenesis, Hr-Erf mRNA was detected from eggs to early developmental stage embryos, whereas the transcript levels were decreased after neurula stage. Similar to the QPCR results, maternal transcripts of Hr-Erf was detected in the fertilized eggs by whole-mount in situ hybridization. Maternal mRNA of Hr-Erf was gradually lost from the neurula stage. Zygotic expression of Hr-Erf started in most blastomeres at the 8-cell stage. At gastrula stage, Hr-Erf was specifically expressed in the precursor cells of brain and mesenchyme. When MEK inhibitor was treated, embryos resulted in loss of Hr-Erf expression in mesenchyme cells, and in excess of Hr-Erf in a-line neural cells. These results suggest that zygotic Hr-Erf products are involved in specification of mesenchyme and neural cells.

• The Journal of Korean Medicine
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• 제19권1호
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• pp.392-409
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• 1998

The present experiment was designed to examine the effects of Jeongjihwan on carecholamines, serotonin, amino acids, lipid peroxide, free radical scavenging activity, malondialdehyde and superoxide dismutase activity in senile brain. It was performed by administering Jeongjihwan extracts of a variety of concentration to senile rats to experimentally determine effects of Jeongjihwan on biochemical changes in senile brain and examine protective effects against neurotoxin. To examine survival rate, the rat's spinal cord sensory ganglion cell pretreated in Jeongjihwan extracts was cultured in oxygen free radical. The results were summarized as follows: 1. Jeongjihwan significantly increased noradrenaline in the hippocampus and hypothalamus of the brain tissue of senile rats, and even though Jeongjihwan increased noradrenaline also in other brain tissue, there was no significance. 2. Jeongjihwan had no effects on dopamine changes in all brain tissue of senile rats. 3. Jeongjihwan significantly increased serotonin, but decreased in other brain tissue. 4. Jeongjihwan increased amino acid in the brain tissue of senile rats. 5. Jeongjihwan significantly decreased lipid peroxide and free radical in the brain tissue of senile rats. 6. Jeongjihwan significantly increased survival rate of nerve cell exposed to oxygen free radical. According to the above results, Jeongjihwan is assumed to improve brain function by reacting on biochemical changes of the senile brain and carries effects of protecting against neurocytotoxicity, and that Jeongjihwan can be used to treat regressive brain disease carrying symptoms of psychoactive disorders.

• Journal of Korean Neurosurgical Society
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• 제43권2호
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• pp.97-104
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• 2008

Objective: Transient receptor potential vanilloid subfamily type 1 (TRPV1), a most specific marker of the nociceptive primary afferent, is expressed in peptidergic and non-peptidergic primary afferents innervating skin and viscera. However, its expression in sensory fibers to skeletal muscle is not well known. In this study, we studied the neurochemical characteristics of TRPV1-positive primary afferents to skeletal muscles. Methods: Sprague-Dawley rats were injected with total $20{\mu}l$ of 1% fast blue (FB) into the gastrocnemius and erector spinae muscle and animals were perfused 4 days after injection. FB-positive cells were traced in the L4-L5 (for gastrocnemius muscle) and L2-L4 (for erector spinae muscle) dorsal root ganglia. The neurochemical characteristics of the muscle afferents were studied with multiple immunofluorescence with TRPV1, calcitonin gene-related peptide (CGRP) and $P2X_3$. To identify spinal neurons responding to noxious stimulus to the skeletal muscle, 10% acetic acids were injected into the gastrocnemius and erector spinae muscles and expression of phospho extracellular signal-regulated kinase (pERK) in spinal cords were identified with immunohistochemical method. Results: TRPVl was expressed in about 49% of muscle afferents traced from gastrocnemius and 40% of erector spinae. Sixty-five to 60% of TRPV1-positive muscles afferents also expressed CGRP. In contrast, expression of $P2X_3$ immnoreaction in TRPV1-positive muscle afferents were about 20%. TRPV1-positive primary afferents were contacted with spinal neurons expressing pERK after injection of acetic acid into the muscles. Conclusion: It is consequently suggested that nociception from skeletal muscles are mediated by TRPV1-positive primary afferents and majority of them are also peptidergic.

• Applied Microscopy
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• 제12권2호
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• pp.55-68
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• 1982

The study on the nerve cells in the pars intercerebralis(IP) of 5-day-old cabbage butterfly Pieris rapae L. was performed to observe their ultrastructures and classify them on the basis. of the differences in size, shape and relative distribution cf cell organelles. The brain-subesophageal ganglion complex was fixed in 1% paraformaldehyde-1% gluaraldehyde mixture and embedded in araldite mixture. The transverse thin sections of IP were stained with uranyl acetate and lead citrate and examined by Hitachi 500 and ]EM 100B electron microscope. Five distinct types. of nerve cells are recognized and are arbitrarily designated as Type I, Type II Type III, Type IV and Type V. Type I neurone: These neurones are neurosecretory cells. Several neurosecretory cells are. recognized in the pars intercerebralis. They are roughly round or peach-shaped cells measuring $13{\sim}25{\mu}m$ in diameter. The rounded nucleus shows about $5{\sim}10{\mu}m$ in diameter. The chromatin is predominantly diffused with only occasional dense patches. The perikaryon contains numerous. mitochondria, free polyribosomes and neurosecretory granules. The neurosecretory granules are relatively uniform in electron density, and each one is about $100{\sim}400{\mu}m$ in diameter and surrounded by a single membrane. The granules are also observed mostly as in groups. In one group of neurones the cisternae of endoplasmic reticulum are distended or in other group of neurones are not distended. Golgi saccules are slightly dilated at their lateral extremities and contains. frequenty dense rounded materials. Type II neurone: Thes have the largest soma in the pars intercerebralis about $30{\sim}35{\mu}m$ in diameter. They also show roughly polygonal in shape. The nucleus is elongated or sickle-shaped. The chromatin is mainly in the euchromatin form. The perikarya in these cells are well populated with populated with free ribosomes and contain numerous mitochondria and Golgi bodies. The cisternae of granular endoplasmic reticulum are also well distributed. Type III neurone: They are oval or spindle-shaped and also medium-sized. neurones approximately $15{\sim}17{\mu}m$ in length. The nucleus is oval or slightly elongated in shape and $8{\sim}9{\mu}m$ in length. The chromatin occurs in diffused form. The cytoplasm contains many filamentous or oval mitochondria. The perikaryon has also numerous free polyribosomes and cisternae of granular endoplasmic reticulum. Type VI neurone: They are roughly polygonal in shape probably due to the close approximation of the adjacent cells. The soma is about $7{\sim}8{\mu}m$ in diameter. The nucleus is round or oval in shape and $5.0{\sim}5.8{\mu}m$ in diameter. The necleus also occupies a large proprion of the cell body. The perikaryon is well populated with free ribosomes and contains several mitochondria and cistenae of granular endoplasmic reticulum. Type V neurone: These neurones are similar to Type VI neurones in various respects such as cell size and cell inclusion, but they differ from Type IV neurones in shape. The soma is oval or slightly elongated. The cell body contains several filamentous and oval mitochondria.

• The Korean Journal of Physiology and Pharmacology
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• 제8권4호
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• pp.219-225
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• 2004

The pelvic ganglia provide autonomic innervations to the various urogenital organs, such as the urinary bladder, prostate, and penis. It is well established that both sympathetic and parasympathetic synaptic transmissions in autonomic ganglia are mediated mainly by acetylcholine (ACh). Until now, however, the properties of ACh-induced currents and its receptors in pelvic ganglia have not clearly been elucidated. In the present study, biophysical characteristics and molecular nature of nicotinic acetylcholine receptors (nAChRs) were studied in sympathetic and parasympathetic major pelvic ganglion (MPG) neurons. MPG neurons isolated from male rat were enzymatically dissociated, and ionic currents were recorded by using the whole cell variant patch clamp technique. Total RNA from MPG neuron was prepared, and RT-PCR analysis was performed with specific primers for subunits of nAChRs. ACh dose-dependently elicited fast inward currents in both sympathetic and parasympathetic MPG neurons $(EC_{50};\;41.4\;{\mu}M\;and\;64.0\;{\mu}M,\;respectively)$. ACh-induced currents showed a strong inward rectification with a reversal potential near 0 mV in current-voltage relationship. Pharmacologically, mecamylamine as a selective antagonist for ${\alpha}3{\beta}4$ nAChR potently inhibited the ACh-induced currents in sympathetic and parasympathetic neurons $(IC_{50};\;0.53\;{\mu}M\;and\;0.22\;{\mu}M,\;respectively)$. Conversely, ${\alpha}-bungarotoxin$, ${\alpha}-methyllycaconitine$, and $dihydro-{\beta}-erythroidine$, which are known as potent and sensitive blockers for ${\alpha}7$ or ${\alpha}4{\beta}2$ nAChRs, below micromolar concentrations showed negligible effect. RT-PCR analysis revealed that ${\alpha}3$ and ${\beta}4$ subunits were predominantly expressed in MPG neurons. We suggest that MPG neurons have nAChRs containing ${\alpha}3$ and ${\beta}4$ subunits, and that their activation induces fast inward currents, possibly mediating the excitatory synaptic transmission in pelvic autonomic ganglia.

• Proceedings of the Korean Institute of Information and Commucation Sciences Conference
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• 한국정보통신학회 2018년도 춘계학술대회
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• pp.584-587
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• 2018

Many viruses including mouse hepatitis virus, corona, measles, and sidbis viruses are known as causative virus of inducing demyelination which means destruction of myelination in nervous system of mice. The purpose of this study is to investigate processing of myelination by co-culture of Schwann cells and neuronal cells and demyelination induced by infection of sindbis virusin rat. Schwann cells and neuronal cells from dorsal root ganglion (DRG) in embryos (E16) of rat were cultured in vitro respectively. The purified neuronal cells with anti-mitotic agents and purified Schwann cells were co-cultured. After that, infection of sindbis virus into this myelinated co-culture system was performed. Myelination and demyelination process were observed using antibody of myelin basic protein meaning presence of myelination.We identified myelination and demyelination processing using antibody of peripheral myelin protein 22 (PMP 22) meaning presence of myelinated neuron. This study was supported by the Basic Research Program through the National Research Foundation (NRF) funded by the Ministry of Science, ICT & Future Planning (NRF-2015R1C1A1A01053484 and 2017R1A2B3005753).

• The Journal of Korean Physical Therapy
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• 제22권3호
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• pp.71-77
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• 2010

Purpose: To determine whether upregulation of inducible nitric oxide synthase (iNOS) transcription and translation is related to radicular pain in a model of lumbar disc herniation. Also, to investigate the temporal changes of mRNA expression of iNOS and the identity of iNOS and transient receptor potential vanilloid (TRPV) 1 channel expression cells in dorsal root ganglion (DRG) of a model of lumbar disc herniation. Methods: A lumbar disc herniated rat model was developed by implantation of the autologous nucleus pulposus, harvested from the coccygeal vertebra of each tail, on the left L5 nerve root just proximal to the DRG. Rats were tested for mechanical allodynia of the plantar surface of both hind paws 2 days before surgery and 1, 5, 10, 20 and 30 days postoperatively. Reverse transcription polymerase chain reaction (RT-PCR) was used to follow iNOS mRNA expression. To stain iNOS and TRPV1 in DRG, an immunohistochemical study was done 10 days after surgery. Results: A significant drop in mechanical withdrawal threshold on the ipsilateral and contralateral hind paws was observed 1 day after surgery and was prolonged to 30 days in rats with lumbar disc herniation. The expression of mRNA for iNOS peaked at postoperative day 10 on both sides of the DRG. iNOS-positive sensory neurons in the DRG varied in size from large to small diameter cells. A majority of small and intermediate sensory neurons were TRPV1-positive cells. Double immunofluorescence staining for TRPV1 and iNOS revealed that most intermediate TRPV1-positive sensory neurons co-localized with iNOS-positive neurons. Conclusion: Nucleus pulposus-induced mechanical allodynia can be generated without mechanical compression. This pain is related to temporal changes in expression of iNOS mRNA in the DRG. Co-localization of TRPV1 and iNOS in intermediate neurons of the DRG is correlated with pain modality and intensity.

• The Korean Journal of Physiology and Pharmacology
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• 제3권1호
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• pp.101-117
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• 1999

Amyotrophic lateral sclerosis (ALS) is a degenerative neuromuscular disease of unknown etiology in which the upper and lower motor neurons are progressively destroyed. Recent evidences support the role of autoimmune mechanisms in the pathogenesis of ALS. This study investigated the effects of sera from ALS patients on neuromuscular transmission in phrenic nerve-hemidiaphragm preparations and on calcium currents of single isolated dorsal root ganglion (DRG) cells in mice. Mice were injected with either control sera from healthy adults or ALS sera from 18 patients with ALS of sporadic form, for three days. Miniature end plate potential (MEPP) and nerve-evoked end plate potential (EPP) were measured using intracellular recording technique and the quantal content was determined. Single isolated DRG cells were voltage-clamped with the whole-cell configuration and membrane currents were recorded. Sera from 14 of 18 ALS patients caused a significant increase in MEPP frequency in normal Ringer's solution $(4.62{\pm}0.14\;Hz)$ compared with the control $(2.18{\pm}0.15\;Hz).$ In a high $Mg^{2+}/low\;Ca^{2+}$ solution, sera from 13 of 18 ALS patients caused a significant increase in MEPP frequency, from $2.18{\pm}0.31$ Hz to $6.09{\pm}0.38$ Hz. Sera from 11 of 18 patients produced a significant increase of nerve-evoked EPP amplitude, from $0.92{\pm}0.05$ mV to $1.30{\pm}0.04$ mV, while the other seven ALS sera did not alter EPP amplitude. In the ALS group, EPP quantal content was also elevated by the sera of 14 patients (from $1.49{\pm}0.07$ to $2.35{\pm}0.07).$ MEPP frequency and amplitude in wobbler mouse were $4.03{\pm}0.53$ Hz and $1.37{\pm}0.18$ mV, respectively, which were significantly higher than those of wobbler controls (wobblers without the symptoms of wobbler). Sera from ALS patients significantly reduced HVA calcium currents of DRG cells to 42.7% at -10 mV. Furthermore, the inactivation curve shifted to more negative potentials with its half-inactivation potential changed by 6.98 mV. There were, however, significant changes neither in the reversal potential of $I_{Ca}$ nor in the I-V curve. From these results it was concluded that: 1) The serum factors of sporadic ALS patients increase neuromuscular transmission and can alter motor nerve terminal presynaptic function. This suggests that ALS serum factors may play an important role in the early stage of ALS, and 2) Calcium currents in DRG cells were reduced and rapidly inactivated by ALS sera, suggesting that in these cells, ALS serum factors may exert interaction with the calcium channel.

• Journal of Acupuncture Research
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• 제17권2호
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• pp.95-117
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• 2000

The purpose of this morphological studies was to investigate the relation between the meridian, acupoints and viscera using neuroanatomical tracers. The common locations of the spinal ganglia, sympathetic chain ganglia, spinal cord and brain projecting to the large intestine meridian were observed following injection of transganglionic tracer, WGA-HRP and transsynaptic neurotropic virus, pseudorabies virus(PRV), Bartha strain(Ba) and PRV-Ba-Gal (Galactosidase)) into the the large intestine(cecum, colon and rectum), ST37 and LI4. After survival times of 96 hours following injection into the thirty rats with WGA-HRP, PRV-Ba and PRV-Ba-Gal. They were perfused, and their spinal ganglia, sympathetic chain ganglia, spinal cord and brain were frozen sectioned($30{\mu}m$). These sections were stained by HRP and X-gal histochemical and PRV immunohistochemical staining method, and observed with a light microscope. The results were as follows : 1. WGA-HRP labeled neurons innervating the large intestine were observed bilaterally within the T13-L4 sympathetic chain ganglia, and T9-11 spinal ganglia. WGA-HRP labeled neurons innervating ST37 were observed within the L3-5 sympathetic chain ganglia, and L2-4 spinal ganglia. WGA-HRP labeled neurons innervating LI4 were observed in the middle cervical ganglion and stellate ganglion, and C5-8 spinal ganglia. 2. In spinal cord, PRV-Ba labeled neurons projecting to the large intestine, ST37 and LI4 were found in thoracic, lumbar and sacral spinal segments. Densely labeled areas of each spinal cord segment were founded in lamina N, V, VII(intermediolateral nucleus), Ⅸ, X and dorsal nucleus. 3. In medulla oblongata, PRV-Ba and PRV-Ba-Gal labeled neurons projecting to the large intestine, ST37 and LI4 were commonly found in the A1 noradrenalin cells/C1 adrenalin cells/caudoventrolateral reticular nucleus, dorsal motor nucleus of vagus nerve, nucleus tractus solitarius, raphe obscurus nucleus, raphe pallidus nucleus, raphe magnus nucleus and gigantocellular nucleus. 4. In pons, PRV-Ba and PRV-Ba-Gal labeled neurons were commonly found in locus coeruleus, Kolliker-Fuse nucieus and A5 cell group. 5. In midbrain, PRV-Ba and PRV-Ba-Gal labeled neurons were commonly found in central gray matter. 6. In diencephalon, PRV-Ba and PRV-Ba-Gal labeled neurons were commonly found in paraventricular hypothalamic nucleus. These results suggest that PRV-Ba and PRV-Ba-Gal labeled common areas projecting to the large intestine may be correlated to that of the large intestine meridian, ST37 and LI4. Especially, These morphological results provide that interrelationship of meridian-acupoints -viscera may be related to the central autonomic pathways.