• The Korean Journal of Physiology and Pharmacology
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• 제6권3호
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• pp.143-147
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• 2002

Granule cells in dentate gyrus of hippocampus relay information from entorhinal cortex via perforant fiber to pyramidal cells in CA3 region. Their electrical activities are known to be closely associated with seizure activity as well as memory acquisition. Since action potential is a stereotypic phenomena which is based on all-or-none principle of $Na^+$ current, the neuronal firing pattern is mostly dependent on afterpotentials which follows the stereotypic $Na^+$ spike. Granule cells in dentate gyrus show afterdepolarization (ADP), while interneurons in dentate gyrus have afterhyperpolarizaton. In the present study, we investigated the ionic mechanism of afterdepolarization in hippocampal dentate granule cell. Action potential of dentate granule cells showed afterdepolarization, which was characterized by a sharp notch followed by a depolarizing hump starting at about $-49.04{\pm}1.69\;mV\;(n=43,\;mean{\pm}SD)$ and lasting $3{\sim}7$ ms. Increase of extracellular $Ca^{2+}$ from 2 mM to 10 mM significantly enhanced the ADP both in amplitude and in duration. A $K^+$ channel blocker, 4-aminopyridine (4-AP, 2 mM), enhanced the ADP and often induced burst firings. These effects of 10 mM $Ca^{2+}$ and 4-AP were additive. On the contrary, the ADP was significantly suppressed by removal of external $Ca^{2+},$ even in the presence of 4-AP (2 mM). A $Na^+$ channel blocker, TTX (100 nM), did not affect the ADP. From these results, it is concluded that the extracellular $Ca^{2+}$ influx contributes to the generation of ADP in granule cells.

• 한국동물학회지
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• 제28권3호
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• pp.194-208
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• 1985

실험에 사용한 플라나리아는 Dugesia japonicia Ichikawa et Kawakatsu로서 核型分析에 의하여 種을 확인하였다. 이 種의 表皮上皮을 구성하는 세포들의 세포화학적 및 미세구조적 특징을 밝히고자 본 실험을 行하였다. 세포화학적 방법으로는 hematoxylin-eosin, periodic acid-schiff(PAS)反應, alcian blue(pH 2.5), PAS-alcian blue反應 그리고 methylene blue-basic fuchsin 등으로 염색을 시행하였고, 체표면을 走査電子顯微鏡으로, 또한 各部位의 微細構造를 透過電子顯微鏡으로 관찰하여 다음과 같은 결과를 얻었다. 플라나리아의 表皮는 單層圓桂形上皮로 되어 있었고, 상피를 구성하는 세포들은 형태학적 및 세포화학적 특성에 의하여 6類型의 세포들도 區分되었다. 1) 纖毛上皮細胞 (Ciliated epithelial cell) : microtubule 배열이 9+2型인 섬모를 가지고 있었다. 2) 好酸性細胞(Eosinophilic cell) : 세포질에 소수의 큰 好酸性 顆粒을 가진 세포로서, 전자 현미경하에서는 顆粒들은 棒狀形이고, 전자밀도가 매우 낮은 基質에 섬유상 구조물이 분산되어 있었다. 3) 粘液細胞(Mucous cell) : 세포질에 PAS 반응에 강한 陽性을 보이는 다양한 모습의 과립($0.8\\times0.3 \\muM$) 들이 충만되어 있었다. 4) 棒狀體形成細胞(Rhabdite-forming cell) : 세포질쪽에 매우 강한 eosinophilia를 나타내는 다양한 크기의 棒狀體顆粒을 한 개 혹은 여러개 가지고 있었으며, 顆粒의 전자밀도는 높았다. 5) A型 好鹽基性顆粒細胞(Basophilic granule cell type A) 背腹部 상피에서 관찰되었고 그 세포질에는 alcian blue(ph2.5)에 강하게 염색되는 $1.5\\times1.0 \\mum$의과립들을 상당히 많이 포함하고 있었다. 과립들은 형태가 球形에 가깝거나 혹은 불규칙형이였고 전자밀도가 높았다. 이類型의 세포들은 柔組織에서 基底膜은 通過하여 表皮上皮로 이동되었다. 6) B型 好鹽基性細胞(Basophilic granule cell type B) 兩側部의 表皮上皮에서 관찰되었으며 세포질에는 PAS 반응에 강한 陽性을 보이는 $0.7\\times0.4 \\mum$의 과립들을 상당히 많이 包含하고 있었다. 전자밀도가 매우 높은 과립들의 형태는 비교적 불규칙하였다. 이類型의 세포들은 역시 柔組織에서 표피로 이동되었다.

• 대한한의학회지
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• 제23권1호
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• pp.120-132
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• 2002

Objectives : The purpose of this investigation was to evaluate the effect of Goomicheongsim-won Extracts on E20 corticells and P7 cerebellar cells exposed to hypoxia, and the effect on neuronal protection by elimination of Rhinoceros unicornis L. and/or Orpiment $As_2S_3$. Methods : P7 cerebellar cells were grown in various concentrations of KM-A, KM-B, KM- C and KM-D. On 7 DIV (day in vitro), cells were exposed to hypoxia (98% $N_2/5%{;}CO_2,{\;}3{\;}hr,{\;}37^{\circ}C$) and normoxia, and then further incubated for 3 days. Neuronal viabilities were expressed as percentages of control. E20 cortical cells were grown in various concentrations of KM-A, KM-B, KM-C, and KM-D. On 7 DIV, cells were exposed to hypoxia and normoxia, and then further incubated for 3 and 7 days. Results : I. The effect of KM-A on neuronal protection was significantly increased P7 cerebellar granule cells and E20 cortical cells on normoxia and hypoxia. 2. The effect of KM-B on neuronal protection was increased P7 cerebellar granule cells on normoxia, but was significantly decreased P7 cerebellar granule cells on hypoxia. The effect of KM-B on neuronal protection was non-significantly increased E20 cortical cells on normoxia and hypoxia. 3. The effect of KM-C on neuronal protection was non-significantly increased P7 cerebellar granule cells on normoxia and hypoxia and was decreased (p=0.058) on hyperconcentration of the extracts in normoxia. The effect of KM-C on neuronal protection was significantly increased P7 cerebellar granule cells and E20 cortical cells on normoxia and hypoxia (10 DIV), and the effect was E20 cortical cells on normoxia (14 DIV), non-significantly increased E20 cortical cells on hypoxia (14DIV). 4. The effect of KM-D on neuronal protection was increased P7 cerebellar granule cells on normoxia but was not on hyperconcentration of the extracts, was significantly decreased on hyperconcentration of the extracts in hypoxia. The effect of KM-D on neuronal protection was significantly increased E20 cortical cells on normoxia and was significantly increased E20 cortical cells increased on hypoxia (10 DIV). Conclusions : Goomicheongsim-won extracts had applicable effect on E20 corticells and P7 cerebellar cells exposed to hypoxia. The effect on neuronal protection by elimination of Rhinoceros unicornis L. and/or Orpiment $As_2S_3$ was changed.

• 한국동물학회지
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• 제23권2호
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• pp.77-88
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• 1980

도룡뇽 피부점액선의 미세구조를 전자현미경으로 관찰하여 다음과 같은 결과를 얻었다. 1. 피부점액선은 선체부와 분비관으로 구성되며, 선체부는 선상피세포와 근상피세포로 구성되었다. 2. 피부점액선의 선체부의 선상피세포는 명세포, 암세포, mitochondria-rich cell, endoplasmic reticulum-developed cell, secretory granule-containing cell 및 vesiculated cells들로 구성되었다. 3. 명세포, 암세포 및 mitochondria-rich cell은 미분화세포, endoplasmic reticulum-developed cell은 분비전기세포, secretory granule-containing cell은 성숙세포 그리고 vesiculated cell은 분비후기세포라고 생각된다.

• 한국동물학회지
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• 제21권1호
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• pp.29-39
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• 1978

양서류 피부 점액선의 선상피세포의 미세구조를 관찰하기 위하여 양서류 피부조직을 2.5% glutaraldehyde-paraformaldehyde(pH7.2) 1% osmium tetroxide에 전후 고정 한후 ethanol과 aceton으로 탈수, Epon 812에 포매 Sorvall MT-2B ultramicrotome으로 ultrathin section을 만들어 uranyl acetate와 lead citrate으로 염색하여 JEOL-100B형 전자현미경으로 관찰하여 다음과 같은 결과를 얻었다. 1. 양서류 피부 점액선은 선상피세포와 근상피세포로 이루어졌다. 2. 양서류 피부 점액선의 선상피세포는 미세구조적인 면에서 종에 따라 여러형의 세포들이 관찰되었다. a. 도롱뇽에서는 암세포와 명세포가 관찰되었다. b. 무당개구리에서는 mitochondria가 많은 세포와 원형의 분비과립을 갖는 세포들이 관찰되었다. c. 맹꽁이에서는 구형의 분비과립을 갖는 세포와 거품모양의 과립괴를 갖는 세포들이 관찰되었다. d. 개구리 피부 점액선은 A형과 B형점액선으로 구분되었고, A형점액선 상피에서는 mitochondria가 많은 세포와 원형의 분비과립을 갖는 세포, B형점액선 상피에서는 mitochondria가 많은 세포와 많은 분비과립을 갖는 세포 및 거의 전 세포질이 endoplasmic reticulum으로 채워진 세포들이 관찰되었다. 3. 위의 사실들로 미루어 양서류 피부점액선의 선상피는 mitochondria가 많은 미분화세포와 점액분비과립을 많이 함유한 성숙세포 및 전 세포질이 거의 rough endoplasmic reticulum으로 채워진 분비후기세포로 구성된다고 생각된다.

• 한국응용약물학회:학술대회논문집
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• 한국응용약물학회 1996년도 춘계학술대회
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• pp.211-211
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• 1996

Changes in glutamate release and uptake on cerebellar cells after the chronic exposure to lead were investigated. Rats were received 0.25% lead acetate in drinking water from the beginning of the pregnancy. The control group was given 0.125% sodium acetate in drinking water. The cerebellar cells from 7 or 8 day-old pups were cultured. Amino acid release from cerebellar granule cells and the glutamate uptake into cerebellar glial cells were measured using HPLC-ECD. Basal glutamate release and NMDA-induced glutamate release didn't show significant difference. However, the other amino acids in the granule cells obtained from lead exposed pups were less released than the control after the stimulation by NMDA (50$\mu$M). SNAP-induced (50$\mu$M) glutamate release was significantly reduced in granule cells prepared from lead exposed pups. The basal glutamate uptake in glial cells didn't show any difference. However, the uptake in glial cells prepared from lead exposed pups was significantly less blocked by PDC (24$\mu$M) compared to the control group. These results indicate that lead exposure to the mother might affect the Excitatory amino acid system during the development of the offspring.

• Applied Microscopy
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• 제18권2호
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• pp.1-20
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• 1988

The species of the slug used in the experiment is Limax flavus L. For identifying the chemical characteristics of the epidermis, granules and mucus-producing cell of this animal is examined with methylene blue-basic fuchsin double stain and PAS-alcian blue reagent. For the ultrastructural research of the epidermal free surface, the epitheial cell and the parenchymal cell are used with scanning electron microscope and transmission elec-tron microscope respectively. I . Epidermal tissue The epidermal tissue of the slug is observed being divided into the dorsal and the ventral side(toot pad) respectively. 1) Dorsal epidermal tissue The dorsal epidermis of the slug is constituted with the simple columnar epithelium and the microvilli are compacted on the epidermal free surface. Two different types of the secretory granules of the neutral and the acid mucus are observed between the epithelial cells, and the neutral mucous granules are highest electron-dense but the acid mucous granules are observed to be electron-lucent. 2) Foot epidermal tissue The Foot epidermis is formed with the taller simple columnar epithelium than the dorsal epidermis and these cells have both a large number of the microvilli and a few number of the large villi. The secretory granules of three different types, which are acid, neutral and mixed mucous granule of two different types are observed between the epithelial cells. The neutral mucous granules are highest electron dense but the acid mucous granules are observed to be electron-lucent. II . Mucous granule-producing cell and mucus-producing cells Seven different types of the granules-producing cell and the mucus-producing cells are observed between the parenchyma. 1) A-type of acid mucous granule-producing cell The electron-lucent granules are largely occupied in the cytoplasm of these cells and then the granules are surrounded by irregular membrane. These electron-lucent granules exhibit alcianophilia with PAS-alcian blue reaction, so these granules are certified to be acid mucopolysaccharide. 2) B-type of acid mucus-producing cell The nucleus and the cytoplasm of these cells are pushed by the acid mucus of the electron-lucent toward the cell membrane. This mucus has been confirmed to be the acid mucopolysaccharide with PAS-alcian blue reagent. 3) A-type of neutral mucous granule-producing cell These cells contain the electron-dense round granules with approximately $1{\mu}m$ in diameter, which exhibit strongly PAS-positive reaction. These granules are confirmed to be the neutral mucoplysaccharide. 4) B-type of neutral mucous granule-producing cell These cells contain two different types of electron dense granules and electron-lucent granules; The former exhibits to be strongly PAS-positive and the latter to have alcianophilia reaction respectively. 5) C-type of neutral mucus-producing cell These cells are similar to the shape and the size of the B-type of mucus-producing cell but these two different types of cells are stained with reversing properties to each other. The mucus of the C-type cell that electron-lucent is largely occupied in the cytoplasm that exhibits strongly PAS-positive reaction. 6) D-type of neutral mucous granule-producing cell These cells contain round granules about $1{\mu}m$ in size which are observed to be medium electron-dense granules and those granules are stained brightly red with PAS-weak positive reaction. The granules are certified to be neutral mucopolysaccharide. 7) E-type of neutral mucous granule-producing cell These cells are similar to the shape and the size of the D-type of neutral mucous granule-producing cell. These cells contain a large number of granules with about $1{\mu}m$ in diameter showing electron-lucent and then granules are seen to be PAS-weak positive reaction. III. Parenchyma The clear cell and dark cell are found in the parenchyma of the Limax flavus L. 1) Clear cell These cells are round formed and the nucleus of the cells are larger than cytoplasm. These cells which have the electron-lucent cytosol possess poorly developed organelles. 2) Dark cell These cells are found to be dark cells due to high electron-density, which exhibit strongly methylene-blue reaction from double stain of methylene blue-basic fuchsin.

• Toxicological Research
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• 제6권2호
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• pp.191-204
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• 1990

Trimethyltin (TMT) induced lesions in the rat hippocampal formation was reviewed. Adult rats were treated with a single dose of 6.0 mg TMT/kg b.w. and were sacrificed between 3-60 days following exposure. On the hippocampal formation, the granule cells of fascia dentata showed early changes which subsided considerably at a later time when the destruction of the pyramidal neurons of the Ammon's horn became increasingly pronounced with time, leading to severe destruction of the structure. It is interesting to note that there was an inverse relationship of pathological involvement between the f.d. granule cells and the Ammon's horn neurons; i.e., when there was a large sparing of the granule cells. there was an extensive damage to the Ammon's horn and vice versa. This inverse relationship was also true between the $CA_3$neurons and the $CA_{1,2}$neurons in the Ammon's horn. Progressive zinc loss, as demonstrated by Timm's method, on the Mossy fibers was also observed. Similar Mossy fiber zinc depletion has been demonstrated in electrical stimulatory excitation condition of the perforant path to the hippocampus. Depletion of corticosterone, an inhibitor to the hippocampal neurons, by means of adrenalectomy will exaggerate the TMT induced hippocampal lesion. Neonatal study revealed that a unique degenerative pattern of the Ammon's horn could be established in accordance with exposure to TMT at specific maturation periods of the fippocampal formation: increasing destruction of the Ammon's horn with increasing synaptogenesis between the f.d. granule cells and the Ammon's horn neurons. Thus it is apparent that the damage of the Ammon's horn, upon exposure to TMT, may depend on the integrity and functional state of the f.d. granule cells. A hyperexcitory scheme and mechanism as the toxicity basis of TMT in the hippocampal formation is proposed and discussed.

• Applied Microscopy
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• 제32권2호
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• pp.175-183
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• 2002

한국 연근해산 살오징어와 서해낙지의 시엽에서 신경전달물질을 분비하는 neuron의 특성 및 기능을 확인하기 위해 rabbit anti-dopamine (TH)과 rabbit anti-calbindin-$D_{28K}$ 등의 항체를 이용한 면역염색과 면역금지법을 시행한 결과는 다음과 같았다. 항-dopamine을 사용한 면역염색 결과 살오징어에서는 외과립세포층 상단에 위치한 대형 무축삭세포와 수질부의 섬을 이루는 세포들 중 $2{\sim}3$개의 비교적 소량의 세포만이 양성반응을 보인 반면, 서해낙지에서는 외과립세포층 상단 및 중간 부위의 $2{\sim}3$개의 세포와 수질부섬을 이루는 5개 이상의 세포에서 강한 양성반응을 보였다. 항-calbindin-D28k인 경우는 살오징어의 외과립세포층 상단에 위치한 $2{\sim}3$개의 소형 무축삭세포와 내과립세포층 하단에 위치한 $1{\sim}2$개의 세포에서 양성반응을 보인 반면, 서해낙지인 경우는 외과립세포층에서 4개 정도의 세포에서 양성 반응을 보였으나, 내과립세포층에서는 반응을 확인할 수 없었다. 면역금지법을 시행한 결과 살오징어에서는 항-dopamine과 항-calbindin에 면역반응을 보인 세포들의 세포질 $0.5{\mu}m^2$당 $17{\sim}26$개 정도의 비교적 많은 수의 금입자가 표지된 반면, 서해낙지에서는 세포질 $0.5{\mu}m^2$당 평균 10개 정도인 소량의 금입자만이 표지되어, 살오징어가 서해낙지에 비해 표지된 금입자가 거의 2배 정도 많았다.

• The Korean Journal of Physiology and Pharmacology
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• 제2권1호
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• pp.41-48
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• 1998

The present study was undertaken to characterize homocysteic acid (HCA)-and cysteic acid (CA)-mediated formation of inositol phosphates (InsP) in primary culture of rat cerebellar granule cells. HCA and CA stimulated InsP formation in a dose-dependent manner, which was prevented by the N-methyl-D-aspartate (NMDA) receptor antagonist D,L-2-amino-5-phosphopentanoic acid (APV). CA-, but not HCA-, mediated InsP formation was in part prevented by the metabotropic glutamate receptor antagonist ?${\alpha}$-methyl-4-carboxyphenylglycine ($({\pm})$-MCPG). Both HCA- and CA-mediated increases in intracellular calcium concentration were completely blocked by APV, but were not altered by $({\pm})$-MCPG. CA-mediated InsP formation was in part prevented by removal of endogenous glutamate. In contrast, the glutamate transport blocker L-aspartic acid-${\beta}$-hydroxamate synergistically increased CA responses. These data indicate that in cerebellar granule cells HCA mediates InsP formation wholly by activating NMDA receptor. In contrast, CA stimulates InsP formation by activating both NMDA receptor and metabotropic glutamate receptor, and in part by releasing endogenous glutamate into extracellular milieu.