• Applied Microscopy
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• 제28권2호
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• pp.139-158
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• 1998

The development of the superior cervical ganglion was studied by electron microscopic method in human fetuses ranging from 40 mm to 260 mm of crown-rump length (10 to 30 weeks of gestational age). At 40 mm fetus, the superior cervical ganglion was composed of clusters of undifferentiated cell, primitive neuroblast, primitive supporting cell, and unmyelinated fibers. At 70mm fetus, the neuroblasts and their processes were ensheated by the bodies or processes of satellite cells. The cytoplasm of the neuroblast contained rough endoplasmic reticulum, mitochondria, Golgi complex, Nissl bodies and dense-cored vesicles. As the neuroblasts grew and differentiated dense-cored vesicles moved away from perikaryal cytoplasm into developing processes. Synaptic contacts between the cholinergic axon and dendrites of postganglionic neuron and a few axosomatic synapses were first observed at 70 mm fetus. At 90 mm fetus the superior cervical ganglion consisted of neuroblasts, satellite cells, granule-containing cells, and unmyelinated nerve fibers. The ganglion cells increased somewhat in numbers and size by 150 mm fetus. Further differentiation resulted in the formation of young ganglion cells, whose cytoplasm was densely filled with cell organelles. During next prenatal stage up to 260 mm fetus, the cytoplasm of the ganglion cells contained except for large pigment granules, all intracytoplasmic structures which were also found in mature superior cervical ganglion. A great number of synaptic contact zones between the cholinergic preganglionic axon and the dendrites of the postganglionic neuron were observed and a few axosomatic synapses were also observed. Two morphological types of the granule-containing cells in the superior cervical ganglion were first identified at 90 mm fetus. Type I granule-containing cell occurred in solitary, whereas type II tended to appeared in clusters near the blood capillaries. Synaptic contacts were first found on the solitary granule-containing cell at 150 mm fetus. Synaptic contacts between the soma of type I granule-containing cells and preganglionic axon termials were observed. In addition, synaptic junctions between the processes of the granule- containing cells and dendrites of postganglionic neuron were also observed from 150 mm fetus onward. In conclusion, superior cervical ganglion cells and granule-containing cells arise from a common undifferentiated cell precursor of neural crest . The granule-containg cells exhibit a local modulatory feedback system in the superior cervical ganglion and nay serve as interneurons between the preganglionic and postganglionic cells.

• 한국패류학회지
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• 제22권1호
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• pp.13-22
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• 2006

2005년 1월부터 12월까지 충청남도 서천군 비인만 앞바다 조간대에서 채집된 큰구슬우렁이를 대상으로 생식소중량지수(GSI), 난자형성 중 난황형성과정 및 생식주기를 세포, 조직학적 관찰과 형태측정 자료에 의해 조사하였다. GSI의 월별 변화는 생식소발달과 유사한 양상을 보였다. 초기 난황형성 단계 난모세포에서 골지체와 미토콘드리아가 지방적 및 난황과립의 형성에 관여되었다. 후기 난황형성 단계의 난모세포에서 조면소포체와 다포체가 세포질 내에서 단백질성 난황과립에 관여하였다. 성숙 난황과립은 주소체 (중앙중심), 가장 자리에 표면층, 그리고 이들을 둘러싼 한계막의 3 가지 성분으로 구성되어있다. 산란기는 6월 초-8월 말 사이 이었고, 주산란은 수온이 $19^{\circ}C$ 이상인 7-8월에 일어났다 암컷의 생식주기는 초기 활성기 (12-2월), 후기 활성기 (2-3월), 완숙기 (4-7월), 산란기 (6-8월), 회복기 (8-11월) 의 연속적인 5 단계로 구분되었다. 완숙난모세포의 크기는 직경이 $250-270{\mu}m$이었다.

• The Korean Journal of Physiology and Pharmacology
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• 제26권4호
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• pp.263-275
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• 2022

There is a paucity of detailed data related to the effect of senescence on the mitochondrial antioxidant capacity and redox state of senescent human cells. Activities of TCA cycle enzymes, respiratory chain complexes, hydrogen peroxide (H₂O₂), superoxide anions (SA), lipid peroxides (LPO), protein carbonyl content (PCC), thioredoxin reductase 2 (TrxR2), superoxide dismutase 2 (SOD2), glutathione peroxidase 1 (GPx1), glutathione reductase (GR), reduced glutathione (GSH), and oxidized glutathione (GSSG), along with levels of nicotinamide cofactors and ATP content were measured in young and senescent human foreskin fibroblasts. Primary and senescent cultures were biochemically identified by monitoring the augmented cellular activities of key glycolytic enzymes including phosphofructokinase, lactate dehydrogenase, and glycogen phosphorylase, and accumulation of H₂O₂, SA, LPO, PCC, and GSSG. Citrate synthase, aconitase, α-ketoglutarate dehydrogenase, succinate dehydrogenase, malate dehydrogenase, isocitrate dehydrogenase, and complex I-III, II-III, and IV activities were significantly diminished in P25 and P35 cells compared to P5 cells. This was accompanied by significant accumulation of mitochondrial H₂O₂, SA, LPO, and PCC, along with increased transcriptional and enzymatic activities of TrxR2, SOD2, GPx1, and GR. Notably, the GSH/GSSG ratio was significantly reduced whereas NAD⁺/NADH and NADP⁺/NADPH ratios were significantly elevated. Metabolic exhaustion was also evident in senescent cells underscored by the severely diminished ATP/ADP ratio. Profound oxidative stress may contribute, at least in part, to senescence pointing at a potential protective role of antioxidants in aging-associated disease.

• Applied Microscopy
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• 제7권1호
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• pp.21-43
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• 1977

This experiment was undertaken in order to localize the labeled dbcAMP (dibutyryl cyclic AMP) in oocytes whose development has been suppressed by cold dbcAMP for 6 or 19 hours in vitro. Mouse oocytes were obtained from the ovaries of 3-4 week old A strain female mice, by puncturing the Graafian follicles in the modified Krebs-Ringer bicarbonate salt solution under the dissecting microscope. Those oocytes which have intact germinal vesicle were cultured in the basic culture medium supplemented with 0.4% bovine serum albumin (BSA). Cultivation of the oocytes was carried out in a microtube developed by Cho (1974). The cultures were then incubated in a humidified 5% $CO_2$ incubator maintained at $37^{\circ}C$ for 6 or 19 hours (Donahue, 1968). DbcAMP was added to culture medium for a final concentration of 100ug/ml, and $^3H-dbc$ AMP (specific activity 13 Ci/mM) for a final concentration of $40{\mu}Ci/ml$ was also added to the medium. For electron microscopic autoradiography, those oocytes recovered from the culture were washed with phosphate buffer (pH 7.4), and immediately prefixed in a 2.5% glutaraldehyde overnight and postfixed for 2 hours at $4 ^{\circ}C$ in 1% osmium tetroxide in phosphate buffer with pH 7.4 (Palade, 1952). After fixation, the materials were dehydrated in graded alcohol series and embedded in Epon 812 mixture based on the standard procedures (Luft, 1961). The thin sections $600-700{\AA}$ thick were mounted on the grids of 200 meshes. The grids containing sections were coated with a nuclear emulsion Kodak NTB-3 and stored in a cold dark box (at $4^{\circ}C$) for 3 weeks. After exposure, the samples were developed with Kodak D-19 and stained with uranyl acetate and lead citrate. Routine observation was made with Hitachi HU-11E electron microsocope. The results of the observation were as followings: 1. It was found that the labeled dbcAMP penetrated the egg plasma membrane and dispersed at random in the cytoplasm. 2. It was also observed that most of the labeled dbcAMP was attached to microfibrillar lattices portion of the oocyte cytoplasm. There fore, it is presumed that the receptor of the dbcAMP is localized in the microfibrillar lattices of the oocyte. 3. It also seems that some other cell organells such as mitochondria, Golgi complex, cortical granules are not directly related to the action of the dbcAMP. 4. The labeled dbcAMP was neither observed in the membrane nor in the nucleus. Therefore, it seems that there is no relationship between the concentration of dbcAMP and the nuclear membranous permeability. 5. There was no difference in number of dbcAMP particles when oocytes were cultured for 6 hours and 19 hours. 6. However, it was observed that, in same of the oocytes suppressed in germinal vesicle by dbcAMP for 19 hours, cell organells were moved and concentrated to a small portion of the cytoplasm, and that the morphology of the organells greatly changed to an abnormal. form. Therefore, it is supposed that those oocytes were in the process of degeneration. From the above results, it is expected that dbcAMP penetrated the egg membrane and was bound to the receptor which seems to be located in the microfibrillar lattiees portion, and that this dbcAMP-receptor complex inhibited some enzyme system of the oocytes which are essential for the germinal vesicle breakdown.

• Applied Microscopy
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• 제30권2호
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• pp.193-204
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• 2000

Mongolian gerbil (Meriones unguiculatus)은 선천적으로 불완전한 뇌바닥동백고리와 특이한 콜레스테롤 대사를 가진 동물로서 수분의 섭취없이도 장기간 생존이 가능하기 때문에 최근 뇌신경학 연구방법에 많이 이용되는 동물이다. 따라서 본 연구는 mongolian gerbil을 절수 시켰을 때 시간의 경과에 따라 뇌에서의 catecholaminergic neuron의 형태적인 변화에 관하여 알아보고자 $50\sim80g$의 mongolian gerbil 25마리를 이용하여 5마리는 뇌도보를 작성하였으며 나머지는 정상군, 5일군, 10일군, 20일군으로 각각 5마리씩 나누어 TH, DBH, PNMT항체를 이용하여 면역조직화학염색하여 절수의 시간에 따른 뇌에서의 catecholaminer-gic neuron의 형태적인 변화를 광학 및 전자현미경적으로 관찰하여 다음과 같은 결론을 얻었다. 1. 정상군의 경우 시상하부의 제 3 뇌실주위에 있는 뇌실곁핵과 뇌실주위회백질에 TH면역반응세포들이 나타났고 이들의 신경세포돌기 및 신경섬유들도 많이 분포하였으나 20일군에서 감소되는 양상을 보였다. 2. 중뇌 및 교뇌에서는 catecholamine성 신경세포중 dopamine성 신경세포가 뚜렷히 관찰되었으나 noradrenalin성 신경세포는 약하게 관찰되었다. 3. 흑색질의 치밀부분과 배쪽피개영역에서 TH항체에 대한 면역반응을 보이는 Dopamine성 신경세포 밀집도가 높았으며 그물부분에서도 소수 관찰되었다. 그러나 이 부위에서 dopamine성 신경세포도 절수에 따라 5일군까지 신경세포체와 돌기 및 섬유들이 감소되었으나 10일군에서는 다시 증가하였으며 20일군에서부터 감소하는 양상을 보였다. 4. 절수에 의한 뇌의 TH면역반응세포체의 전자현미경적 소견은 정상군에 비해 5일군에서 핵의 농축 및 변형, 신경연접의 소실, 사립체의 감소, 축삭 수의 감소 등이 나타나 TH의 분비에 영향을 주는 것으로 보인다. 그러나 ER등의 변화는 크게 관찰할 수 없었다. 이러한 연구결과는 정상군의 경우 다른 동물과 유사했으나 다른 동물에서는 청색반점이나 교뇌부위에서 noradrenaline성 신경이 많다고 하였지만 본 연구에서는 그러한 차이를 발견하지 못했으며 이것은 동물간의 유전적, 환경적 차이로 사료된다. 그러므로 mongolian gerbil의 수분대사에 dopamine이 관여할 것으로 보고 탈수 질환모델로 개발하기 위한 기능적인 연구와 더불어 많은 형태학적인 연구가 병행되어야 할 것이다.

• Applied Microscopy
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• 제29권4호
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• pp.479-491
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• 1999

출생후 0일, 7일, 14일, 17일, 21일, 28일, 35일 그리고 성체의 흰쥐 전뇌 기저부의 Meynert 기저핵에서 ChAT 면역반응 신경세포의 분화를 면역조직화학적 및 전자현미경적 방법을 이용하여 조사하였다. 출생후 7일에서 성체까지 신경세포의 세포질에서 ChAT면역반응이 확인되었다. 특히 ChAT면역반응은 세포체의 세포질과 수상돌기에 고루 분포하였다. 전뇌 기저부의 ChAT면역반응 신경세포들은 발생에 따른 뇌 크기의 증대와 뇌조직의 분화에 따라 점차 수적 증가를 보였다. ChAT면역반응 신경세포들은 세포의 모양과 세포체의 장 단축의 비에 따라 6가지 형으로 분류되었다. 성체의 Meynert기저핵에서 원형은 9.4%, 난원형은 35.5%, 세장형은 32.1%, 방추형은 5.9%, 삼각형은 9.1% 그리고 다각형은 8.0%의 출현율을 보였다. 원형과 난원형 신경세포들의 출현율은 출생후 14일에서 가장 높아 각각 18.7%, 55.5%였고, 성체로 되면서 점차적으로 감소되었다. 또한, 세장형과 삼각형 신경세포들의 출현율은 출생후 21일에 가장 높아 각각 30.4%, 10.1%였고, 성체로 되면서 원형과 난원형 세포에서와 같이 점차적으로 감소하였다. 그러나 이들과는 다르게 방추형과 다각형 신경세포들의 출현율은 출생후 발생이 진행됨에 따라 점진적으로 증가하는 현상을 보였다. ChAT면역반응 신경세포체의 부피는 출생후 7일에 $1,083{\mu}m^3$로 제일 작았으나, 출생후 21일에서는 $5,045{\mu}m^3$로 최대치를 보였다. 그후 성체로 되면서 $2,731{\mu}m^3$로 감소되었다. 전자현미경적 관찰에서 출생후 21일의 경우 ChAT면역반응은 신경세포의 ribosome, polysome 그리고 RER에서 관찰되었으며, 대칭 및 비대칭 신경연접이 관찰되었다. 이상의 결과들로 미루어 흰쥐 전뇌 기저부 Meynert 기저핵에서의 ChAT면역반응 신경세포들은 출생후 발생과정에서 세포소기관과 신경돌기들의 분화에 따른 세포체부피의 증가, 분화된 세포형들의 출현율의 증가 및 세포의 손실이 없는 상태에서의 세포 응축 등의 과정을 통하여 세포들이 분화한다고 생각된다. 또한, Golgi체와는 다르게 ribosome, polysome그리고 RER들이 ChAT의 세포내 분포 및 생합성과 밀접한 관계를 같는 것으로 생각된다.

• Applied Microscopy
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• 제15권2호
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• pp.31-68
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• 1985

Authors are trying to unveil the ultrastructural organization of olfactory bulb, which has been summerized under light microscopic level or communicated only in some detail in different view point until now. For the critical point of view, since the phylogenetical approach will give the ultimate value in the correlative study between structural and functional bases (Brodal, 1969), the present study was carried out light and electron microscopic analyses of the structures of the neurons and synaptic organizations in olfactory bulbs from different animals in phylogenetical scale. We selected each one species from five animal classes: the house rabbit(Oryctolagus cuniculus var. domesticus [Gmelin]) from Mammalia, the domestic fowl (Gallus gallus domesticus Brisson) from Aves, the viper (Agkistrodon hylys [G.P. Pallas]) from Reptilia, a frog (Bombiana orientalis Boulenger) from Amphibia and the crussian carp (Carassius carassius [Linne]) from Pisces. For light microscopic study, samples were fixed in 10% formalin and paraffin sections were stained with hematoxylin-eosin. For the electron microscopic study, the tissues were fixed by perfusion through the heart or immersion with 1% paraform-aldehyde-glutaraldehyde mixture (phosphate buffer, pH 7.4), and final tissue block trimmed under dissecting microscope were osmicated (1% OsO4), they were embedded in Araldite or Epon 812, and ultrathin sections were made by LKB-V ultratome following the inspection of semi-thin sections stained with toluidine blue-borax solution. Ultra-thin sections contrasted with uranyl acetate and lead citrate were observed with JEM 100CX electron microscope. We have summerized our morphological analyses as follows: 1. The olfactory bulb of rabbit, viper and frog shows the eight layers of fila olfactoria, glomerular, external granular, external plexiform, mitral cell, internal plexiform, internal granular, medullary but domestic fowl shows the five layers of glomerular, fibrillar, mitral, granular and medullary and the three layers of fibrilla, glomerular and medullary in crussian carp. The sharpness of demarcation between the layers shows deferential tendency according to phylogenetical order. 2. Mitral cells of vertebrate have large triangular or oval shape with spherical nuclei which contain not so much chromatin. The cytoplasm contains numerous cell organelles, of which Nissl's bodies or granular endoplasmic reticula arranged as parallel strands. Development of granular endoplasmic reticula were declined as the phylogentical grade is going lower. 3. Tufted cells of all animal are mostly spindle or polygonal contour and contain oval nuclei which located in periphery of cytoplasm. The nuclei of rabbit, fowl, viper and frog has relatively space chromatin, but a nucleus of crussian carp contain irregularly aggregated chromatin in karyoplasm. Their cytoplasmic volume and cell organelle contents are in between those of mitral cell and granular cell. They contain moderate amount of mitochondria, granular endoplasmic reticula, a few Golgi complex, polysomes, lysosome, etc. 4. Granule of cells of all the vertebrate amimals studied exhibit similar features; cells and their dense nuclei show spherical or oval contour, and they have the thin rim of cytoplasm which contain only a few cell organelles. 5. In rabbit, the soma of mitral cells were in contact with boutons with two types of synaptic vesicles, that is, round and flat vesicles, especially flat vesicles in boutons were showing reciprocal synapses. However, in domestic fowls, vipers, frogs and crussian carps, there were found boutons showing only spherical synaptic vesicles. 6. The boutons containing round synaptic vesicles were made contact with the some of tufted cell of olfactory bulb in the rabbits, fowls, vipers and frogs, but no synaptic boutons were observed in soma of tufted cells in crussian carps. In the frogs, there were observed dendrites were contact with the soma of tufted cells. 7. In the neuropils of plexiform, granular and glomerular layers olfactory bulbs in the vertebrate, the synapses were axo-large dendrites, axo-median and small dendrites, dendrodendritic, and axo-axonal contacts. However, in the neuropil of crussian carps, synapses were observed only in glomerular layer.

• Parasites, Hosts and Diseases
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• 제29권3호
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• pp.245-258
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• 1991

요꼬가와흠흡에 감염된 숙주에서 항체생성을 유도하는 물질의 충체 래에 분포하고 있는 부위를 확인하기 위하여 대조군의 실험동물 IgG와 요꼬가와흡충에 감염된 실험고양이의 특이항체 IgG를 요꼬가와흡충 성충조직에 반응시키고 면역황금표식법을 이용하여 전자현미경으로 관찰하여 다음과 같은 결과를 얻었다. 대조군의 IgG를 반응시킨 조직항원에는 환금입자의 표지가 관찰되지 않았으나 표피층은 표피세포들의 원형질돌기들이 표피합포체를 구성하고 있었으며 기저층은 분명하게 구별되고 경육층도 잘 발달되어 있었다. 난황세포는 크기가 다양한 분필과립들을 포함하고 있었으며 조면소포체가 특히 발달되어 있었다. 맹관의 막구조물은 잘 발달되었으며 장상피 합포체도 잘 발달되어 있었다. 저정낭에는 정자가 전자밀도가 릎은 두부와 미세소관이 잘 발달한 미부로 구성되어 있었으며, 배설낭은 배설낭 상피에 막구조물이 관찰되었고 충란은 전자밀도가 높은 난각이 관찰되었다. 요꼬가와흡충에 감염된 실험동물의 IgG를 반응시킨 조직항원의 표피충에는 표피합포체와 표피세포의 세포질에 황금입자가 높은 밀도로 표지되어 강한 항원성을 나타내었다. 그리고 난황세포의 분필과립과 맹관 막구조물과 맹관 내강에도 환금입자가 높은 밀도로 관찰되어 항원성이 강한 것으로 생각되었다. 저정낭의 정자와 배설낭의 막구조물에는 적은 수의 황금입자가 표지되어 항원성이 미약한 것으로 생각되었으나, 자궁의 충란 난각에는 황금입자가 다소 표지되어 항원성물질이 존재하는 것으로 생각되었다. 이상의 결과로 요꼬가와홉충에 감염된 실험 고양이는 요꼬가와흡충의 표피합포체와 표피세포에서 생성된 물질과 난황세포에서 생성된 물질에 의하여 면역항체가 형성되며 일부는 맹관 내용물에 의하여 면역항체가 유도되는 것으로 생각되었다.는 69.7%의 같은 비율로 높았다. 이상으로 요충 감염은 아직까지 높은 상태임을 알았고 예방을 위한 집단 구충에는 전 집단에 대한 3회 이상의 반복구충이 효과적이었다. gas layers is also made. Low resolution spectral intensities at the boundary are obtained for uniform, parabolic and boundary layer type temeprature profiles using the obtained for uniform, parabolic and boundary layer type temperature profiles using the obtained WSGGM's with 9 gray gases. The results are compared with the narrow band spectral intensities as obtained by a narrow band model-based code with the Curtis-Godson approximation. Good agreement is found between them. Local heat source strength and total wall heat flux are also compared for the cases of Kim et al, which again gives promising agreement.면적 306~453$\textrm{cm}^2$, 유색계의 경우 수당면적 340~453$\textrm{cm}^2$ 일 때 경제능력을 제대로 발휘할 수 있고 경제성이 있는 것으로 나타났다. 첨가구가 높은 경향이 있었다8.4%. 79.7% 그리고