• Korean Journal of Veterinary Research
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• v.39 no.1
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• pp.34-44
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• 1999

Ischemic damage in the selectively vulnerable populations of neurons is thought to be caused by an abnormal accumulation of intracellular calcium. It has been reported that the neurons, expressing specific calcium binding proteins, might effectively control intracellular calcium concentrations because of a high capacity to buffer intracellular calcium in the brain ischemic condition. It is uncertain that parvalbumin, one of the calcium binding proteins, can protect the neurons from the cerebral ischemic damage. Recently, treatment of kappa opioid agonists increased survival rate, improved neurological function, and decreased tissue damage under the cerebral ischemic condition. Many evidences indicate that these therapeutic effects might result from regulation of calcium concentration. This study was designed to analyze the changes of number in parvalbumin-positive neurons after cerebral ischemic damage according to timepoints after cerebral ischemic induction. In addition, we evaluated the effect of GR89696 (kappa opioid agonist) or naltrexone(non selective opioid antagonist) on the changes of number in parvalbumin expressing neurons under ischemic condition. Cerebral ischemia was induced by occluding the common carotid artery of experimental animals. The hippocampal areas were morphometrically analyzed at different time point after ischemic induction(1, 3, 5 days) by using immuno-histochemical technique and imaging analysis system. The number of parvalbumin-positive neurons in hippocampus was significantly reduced at 1 day after ischemia(p<0.05). Furthermore, the number of parvalbumin-immunoreactive neurons was dramatically reduced at 3 and 5 days after cerebral ischemic induction(p<0.05) as compared to 1 day group after ischemia, as well as sham control group. Significant reduction of parvalbumin positive neurons in CA1 region of hippocampus was observed at 1 day after cerebral ischemic induction. However, significant loss of MAP2 immunoreactivity was observed at 3 day after cerebral ischemia. The loss of parvalbumin-positive neurons and MAP2 immunoreactivity in CA1 region was prevented by pre-administration of GR89696 compared to that of saline-treated ischemic group. Furthermore, protective effect of GR89696 partially reversed by pre-treatment of naltrexone. These data indicate that parvalbumin-positive neurons more sensitively responded to cerebral ischemic damage than MAP2 protein. Moreover, this loss of parvalbumin-positive neurons was effectively prevented by the pretreatment of kappa opioid agonist. It was also suggested that the changes of number in parvalbumin-positive neurons could be used as the specific marker to analyze the degree of ischemic neuronal damage.

• 한국생물공학회:학술대회논문집
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• 2003.04a
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• pp.743-746
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• 2003

The major pathological lesion in Parkinson's disease(PD) is selective degeneration and loss of pigmented dopaminergic neurons in substantia nigra (SN). Although the initial cause and subsequent molecular signaling mechanisms leading to the dopaminergic cell death underlying the PD process is elusive, the potent neurotrophic factors (NTFs), brain derived neurotrophic factor (BDNF) and glial cell line derived neurotrophic factor (GDNF), are known to exert dopaminergic neuroprotection both in vivo and in vitro models of PD employing the neurotoxin, MPTP. BDNF and its receptor, trkB are expressed in SN dopaminergic neurons and their innervation target. Thus, neurotrophins may have autocrine, paracrine and retrograde transport effects on the SN dopaminergic neurons. This study determined the BDNF secretion from SN dopaminergic neurons by ELISA. Regulation of BDNF synthesis/release and changes in signaling pathways are monitored in the presence of free radical donor, NO donor and mitochondrial inhibitors. Also, this study shows that BDNF is able to promote survival and phenotypic differentiation of SN dopaminergic neurons in culture and protect them against MPTP-induced neurotoxicity via MAP kinase pathway.

• Toxicological Research
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• v.16 no.1
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• pp.63-66
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• 2000

The effects of acute ethanol on the high K+ induced $Ca^{2+}}$ signals were examined from primary cultures of cerebellar granule neurons. $Ca^{2+}}$ signals were measured with Calcium Green-1 based microscopic video imaging. Because $Ca^{2+}}$ signal was low in most of granule neurons without stimuli, high KCI was used for depolarization. In most case, acute exposure to ethanol reduced the peak amplitude of the $Ca^{2+}}$ signals, induced by high K+, even though low concentration of ethanol(2~10mM) was used and the effects lasted more than 30min. In was also possible to see differences of ethanol inhibition, i.e. the temporal pattern of $Ca^{2+}}$ signal reductions and the strength of inhibition of $Ca^{2+}}$ signals in cerebellar granule neurons. These results indicate that low concentration of ethanol has diverse actions on the $Ca^{2+}}$ signals in cerebellar granule neurons.

• The Korean Journal of Zoology
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• v.36 no.2
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• pp.209-222
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• 1993

The present study attempts to explore the first appearance 8nd subsequent development of %cosine hvdroxvlase (TH)-containing neurons in the rat brain from embryonic day (E) 10.5 to the neonate. To increase the senti노ivity and resolution power, a double bridge peroxidase-anti-peroxidase technique was employed for cellular localization of TH. In situ hybridization histochemistw with synthetic TH oligomer (30-mer) codinB TH was also used to detect TH mRNA. TH-containing neurons were first detected at E11.5 in the intermediate zone of prosencepha1on and mesencephalon. At this stage, TH-immunoreactive neurons were small, ovoid bee and emitted their fibres into their immediate surroundings. From this stage, TH-immunoreactive neurons increased in their number and underwent migration and cell differentiation. At E15.5, the distribution pattern of the maior groups of TH neurons was similar to that of adult catecholaminergic groups, and at E19.5 the external laver of median eminence showed TH-immunoreactive processes.

• Applied Microscopy
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• v.11 no.1
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• pp.1-9
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• 1981

The study on the nerve cells in the subesophageal ganglion 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 of cell organelles. 1. Type I neurons: These cells are neurosecretory granules ranging 100 to 300 nm in size. 2. Type II neurons: As giant neurons averaging 25 to $30{\mu}m$ in size, such as mitochondria and Golgi apparatus. 3. Type III neurons: These spindle-shaped cells range 9 to $15{\mu}m$ in width. 4. Type IV neurons: These cells have a range of diameter from 12 to $16 {\mu}m$. The cells are abundantly observed in the subesophageal ganglion. 5. Type V neurons: These cells are very small nerve cells with 4.5 to $8.0{\mu}m$ in size and have a prominent nucleus.

• Journal of the Korean Institute of Telematics and Electronics B
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• v.31B no.9
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• pp.133-141
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• 1994

Conventional competitive learning algorithms compute the Euclidien distance to determine the winner neuron out of all predetermined output neurons. In such cases, there is a drawback that the performence of the learning algorithm depends on the initial reference(=weight) vectors. In this paper, we propose a new competitive learning algorithm that dynamically generates output neurons. The proposed method generates output neurons by dynamically changing the class thresholds for all output neurons. We compute the similarity between the input vector and the reference vector of each output neuron generated. If the two are similar, the reference vector is adjusted to make it still more like the input vector. Otherwise, the input vector is designated as the reference vector of a new outputneuron. Since the reference vectors of output neurons are dynamically assigned according to input pattern distribution, the proposed method gets around the phenomenon that learning is early determined due to redundant output neurons. Experiments using speech data have shown the proposed method to be superior to existint methods.

• The Journal of Korean Medicine
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• v.21 no.1
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• pp.29-39
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• 2000

In order to elucidate the toxic mechanism of oxygen radicals in cultured mouse spinal sensory neurons, cytotoxic effect of oxygen radicals was evaluated by M1T assay and NR assay. In addition, protective effect of Sopunghwalhyultang(SPHHT) water extract on oxidant-induced neurotoxicity was investigated on these cultures. Spinal sensory neurons derived from mice were cultured in mediums containing various concentrations of Xanthine Oxidase / Hypoxanthine(XO/HX). Cell viability was measured by MTT assay and NR assay. XO/HX-mediated oxygen radicals remarkably decreased cell viability of cultured spinal sensory neurons in a dose-and time-dependent manner. And also, SPHHT blocked XO/HX-induced neurotoxicity in these cultures. These results suggest that oxygen radicals are toxic and SPHHT are effective in blocking against the oxidant-induced neurotoxicity in cultures of spinal sensory neurons of mice.

• The Korean Journal of Zoology
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• v.35 no.4
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• pp.428-438
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• 1992

The serotonin-immunoreactive (5-HTil neurons have been investigated in the brains of lanra, pupa and adult from Pieris ropae. There are ca. 54 5-HTi neurons in 5-instar larva, ca. 20 in 2-dav-old pupa and ca. 118 in 1-day-old adult, respectively. Most of these 5-HTi neurons are interneurons, but efferent and afferent 5-HTi neurons were also observed. Most of the 5-HTi neurons project into the central neuropils of postembrvonic brains. The larval brain contains abundant 5-HTi processes in the central neuropils, including those in three cerebral commissures. But in the pupal brain the 5-HTi processes are restricted in small numbers to the given regions of central neuropil. The adult brain contains a large number of 5-HTi processes in mushroom body, central body complex, lateral protocerebrum, protocerebral bridge, antennal lobe, and tritocerebral and suboesophageal neuropils. However, the 5-HTi processes are not found in the optic lobe of the brains. One prominent feature of the 5-HTi fibers in the postembrvonic brains is the fact that they are greatly arborized.

• International Journal of Oral Biology
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• v.30 no.2
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• pp.71-76
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• 2005

The mesencephalic trigeminal nucleus (Mes V) contains cell bodies of primary afferent sensory neurons that relay proprioceptive information from the periodontium and masticatory muscles and function as typical sensory neurons or potentially as integrative interneurons. In the present study, we studied these two potential functions using combined experimental approaches of retrograde labeling and whole cell patch clamp recording. Mes V neurons that presumably originate from periodontal nerve fibers in subsets of Mes V nucleus were identified by retrograde labeling with a fluorescent dye, DiI, which was applied onto inferior alveolar nerve. These cells were elliptical perikarya shaped cells about $40{\mu}m$ in diameter. In these neurons, we measured high voltage-activated calcium channel (HVACC) currents. $GABA_B$ agonist, baclofen, inhibited calcium currents, and the HVACC currents inhibition by baclofen was voltage-dependent, exhibited prepulse facilitation, indicating that it was mediated by $G_i/_G_o$ protein. Taken together, our results demonstrate that Mes V neurons not only have cell bodies originating from periodontium, but also receive synaptic inputs including GABAergic neurons suggesting that Mes V neurons function as both primary sensory neurons and integrative interneurons.

• The Korean Journal of Physiology and Pharmacology
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• v.12 no.6
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• pp.315-321
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• 2008

Eugenol is widely used in dentistry to relieve pain. We have recently demonstrated voltage-gated $Na^+$ and $Ca^{2+}$ channels as molecular targets for its analgesic effects, and hypothesized that eugenol acts on $P2X_3$, another pain receptor expressed in trigeminal ganglion (TG), and tested the effects of eugenol by whole-cell patch clamp and $Ca^{2+}$ imaging techniques. In the present study, we investigated whether eugenol would modulate 5'-triphosphate (ATP)-induced currents in rat TG neurons and $P2X_3$-expressing human embryonic kidney (HEK) 293 cells. ATP-induced currents in TG neurons exhibited electrophysiological properties similar to those in HEK293 cells, and both ATP- and $\alpha$, $\beta$-meATP-induced currents in TG neurons were effectively blocked by TNP-ATP, suggesting that $P2X_3$ mediates the majority of ATP-induced currents in TG neurons. Eugenol inhibited ATP-induced currents in both capsaicin-sensitive and capsaicin-insensitive TG neurons with similar extent, and most ATP-responsive neurons were IB4-positive. Eugenol inhibited not only $Ca^{2+}$ transients evoked by $\alpha$, $\beta$-meATP, the selective $P2X_3$ agonist, in capsaicin-insensitive TG neurons, but also ATP-induced currents in $P2X_3$-expressing HEK293 cells without co-expression of transient receptor potential vanilloid 1 (TRPV1). We suggest, therefore, that eugenol inhibits $P2X_3$ currents in a TRPV1-independent manner, which contributes to its analgesic effect.