• Asian-Australasian Journal of Animal Sciences
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• 제21권8호
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• pp.1220-1225
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• 2008

This study was conducted to investigate the protective effect of genistein on the antioxidative defence system and membrane fluidity in chick skeletal muscle cells after supplementation with 0, 20, 40, and $80{\mu}mol/L$ genistein in $50{\mu}mol/L$ $FeSO_4/H_2O_2$ treated cells for 24 h. Genistein supplementation recovered the decreased activity of total superoxide dismutase induced by $FeSO_4/H_2O_2$, significantly increased glutathione peroxidase activity (p<0.05) and decreased malondialdehyde production (p<0.05). The treatment of 80 mol/L genistein in $FeSO_4/H_2O_2$ treated cells decreased the secretion of creatine kinase (p<0.05). Fluorescence polarization values and microviscosities observed with $FeSO_4/H_2O_2$ treated cells were significantly higher than those observed with no $FeSO_4/H_2O_2$ treated cells. The addition of $80{\mu}mol/L$ genistein improved the increased fluorescence polarization value (p<0.05) caused by $FeSO_4/H_2O_2$ treatment. The microviscosity value was significantly decreased by adding genistein (p<0.05). In conclusion, genistein protected skeletal muscle cells from oxidative damage by improving antioxidative status and membrane fluidity.

• Applied Microscopy
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• 제20권1호
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• pp.17-35
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• 1990

Effect of 5-hydroxytryptamine(5-HT, serotonin) and its precursor tryptophan on the cell proliferation of brain and somite parts of 4 day chick embryo in Dulbeco's modified essential medium was examined morphologically at cellular level. It was realized that the externally added 5-HT and/or tryptophan disturbed cell proliferation and severve necrosis occured. Electron micrograph showed that the development of cell organelles were greatly impaired. The activities of both acetylcholine esterase and $Mg^{2+}$ -dependent ATPase of the brain tissues of 5 day chick embryo, which received 1mg of tryptophan and/or 0.1mg of 5-HT at primitive streak stage after 24 hrs incubation of the fertilized egg, were much lower(about 20-25%) than those of control group. These results were supported by the electron micrographs of chemically treated cells. Control cells showed clear densed bands of acetylcholine esterase activity around nucleus and rough endoplasmic reticulum but tryptophan or 5-HT treated groups showed discontinued activity bands. In the case of $Mg^{2+}$-ATPase, the control groups showed clear continuous activity bands but tryptophan and/or 5-HT treated groups were discontinuous. From the previous and present studies, it seems that the intracelluar 5-HT level is very important for the cell proliferation and normal morphogenesis.

• Applied Microscopy
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• 제21권2호
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• pp.1-13
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• 1991

In order to investigate the factors of the control mechanism of somitogenesis, early chick embryos (H-H stage $8{\sim}13$) were treated with heat shock, ${\alpha}$-amanitin and cycloheximide and morphological changes of somite were examined by light microscopy, transmission and scanning electron microscopy. In normal chick embryo, somites were formed from the somitomere which preexisted in segmental plate. Somites were wrapped with extracellular collagen fibrils and connected with neural tube, notochord and ectoderm. And then, somites were differentiated to sclerotome, dermatome and myotome by the interaction of nervous tissue. Abnormal somites were observed after formation of six or seven so mites in heat shock treated group. Amounts of collagen fibrils were obviously decreased in this group. In cycloheximide treated group, most so mites were smaller and neural tube formation was incomplete. Chromatins were condenced and formed several heterochromatins in the nucleus of somite cells. Lipid like cytoplasmic dense mass and lipid droplets were also observed. Segmentation of somites seemed to be normal progress in ${\alpha}$-amanitin treated group. Center of somite, however, hollowed in longitudinal sectioned samples. These results suggested that so mites were already existed in the segmental plate as the form of somitomere. Segmented somites were contacted with neural tube or notochord and the somites were tightly connected with each other by the extracellular collagen fibrils which were secreted from neuroepithelium and somite cells. Somites are thought to differentiate into sclerotome, dermatome and myotome by these interactions.

• Applied Microscopy
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• 제26권2호
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• pp.123-136
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• 1996

To investigate the effect of aflatoxin $B_1$, on survival rate and ultrastructure of liver during chick embryogenesis electron microscopic methods were used. After injection of aflatoxin $B_1$ into the yolk, ultrastructural changes in the liver of chicken embryo were observed. The results were as followed. 1. 12-day old chicken embryos were treated with single injection of aflatoxin $B_1$ with the dose of $0.0005{\mu}g,\;0.005{\mu}g,\;0.05{\mu}g,\;0.5{\mu}g,\;2.5{\mu}g,\;5.0{\mu}g$ each. Chicken embryos treated with the dose of $0.5{\mu}g$ of aflatoxin $B_1$ had survival rate of 22%. The embryos treated with $2.5{\mu}g$ of aflatoxin $B_1$ hardly survived. 2. Chicken embryos treated with $0.05{\mu}g$ of afatoxin $B_1$ had hatched in 30%, but once hatched, they all survived. 3. After administration of $0.05{\mu}g$ of aflatoxin $B_1$ into the 12-day old chicken embryo, the electron microscopic studies were examined during development stages. The nuclei of hapatocytes became irregularly shaped and the structures of endoplasmic reticulum were changed to spherical types at 20-day old chicken embryo. Also, mitochondria became to be dilated and severe fibrosis was induced in the cytoplasm. However, the hepatocytes became almost normal in 30-day old young chicken.

• Applied Microscopy
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• 제26권3호
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• pp.253-266
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• 1996

To investigate the effects of mercuric chloride ($HgCl_2$) on the differentiation of the cerebral neuron of chick embryo 9 days, the ultrastructural changes in nerve cells injected with a various doses of mercuric chloride were observed with transmission electron microscope. The enzyme activity of the some dehydrogenases, cerebral proteins and adenosine triphosphate (ATP) were also analyzed. The results obtained are as follows: The ultrastructural changes in 0.5 and 1.0mg-injected groups were undetectable, but in 2.0mg-injected group, the nuclear envelops were very irregular and mitochondria, were swelled and destroyed partly. The number of polypeptide bands separated by SDS-PAGE in the normal group were 37 bands. According to the in creased dose of mercuric chloride, contends of the bands were increased in 7 bands. The activities of dehydrogenases were declined by increasing the dose of mercuric chloride. Lactate dehydrogenase (LDH) activity failed to 78% in 1.0mg-injected group and greatly to 68% in 2.0 mg-injected group. Malate dehydrogenase (MDH) activity failed to 81% in 2.0 mg-injected group. On the other hand, succinate dehydrogenase (SDH) activity decreased to 80% in 1.0 mg-injected group and greatly to 63% in 2.0 mg-injected group. ATP content in 1.0 mg-injected group was increased slightly and in 2.0 mg-injected group was increased greatly.

• Applied Microscopy
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• 제27권1호
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• pp.87-100
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• 1997

To investigate the effects of mercuric chloride $(HgCl_2)$ on the differentiation of the cerebral neuron of chick embryo 10 days, the ultrastructural changes in nerve cells injected with a various doses of mercuric chloride were observed with transmission electron microscope. The enzyme activity of the some dehydrogenases, cerebral proteins and adenosine triphosphate (ATP) were also analyzed. The results obtained are as follows; The ultrastructural changes in 1.0 mg-injected group, the nuclear membranes were irregular, outer of mitochondria membrances dispressioned, their cristae were destroyed. In 2.0 mg-injected group, the nuclear envelops were destroyed and divided, were not observed organelle except of few ribosome, the RER and mitochondria. The number of polypeptide bands were separated by SDS-PAGE in the normal group were 38 bands. According to the in creased dose of mercuric chloride, contends of the bands were increased in 4 bands, but were decreased in 1 band. The activities of dehydrogenases were declined by increasing the dose of mercuric chloride. Lactate dehydrogenase (LDH) activity fatted to 61% in 2.0 mg-injected group. Malate dehydrogenase (MDH) activity fatted to 90% in 1.0 mg-injected group, greatly to 76% in 2.0 mg-injected group. Succinate dehydrogenase (SDH) activity decreased to 79% in 1.0 mg-injected group and greatly to 62% in 2.0 mg-injected group. ATP content in 1.0 mg-injected group was almost near to the normal level, but it was increased greatly in 2.0 mg-injected group.

• Applied Microscopy
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• 제18권1호
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• pp.60-76
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• 1988

Chick embryos which have received a single injection of the organophosphate compound, malathion (0.1 mg/0.05 ml, 0.5 mg/0.05 ml, 1.0 mg/0.05 ml or 2.0 mg/0.05 ml) via the yolk sac at certain times (2 days, 4 days or 6 days after incubation) have been investigated. After 9 days of incubation, chick embryos were harvested to examine the effects of malathion on the ultrastructure and the acetylcholinesterase(AChE) activity of the developing spinal cord. The effects of simultaneous injection of malathion and nicotinamide were also compared. On ultrastructural findings, neurons in the ventral horn of spinal cord showed to be inhibited in their differentiation by malathion; nuclear irregularity, separation of nuclear membranes, reduction of ribosomal distribution, and cytoplasmic vacuoles were observed. In the younger embryos treated with relatively high doses of malathion, nucleus and cytoplasmic organelles of neurons were severely destroyed, and the neurons were shown to be necrotic. On cytochemical study of AChE by electron microscope, the positive reaction products of AChE were localized at the membranes of nucleus and endoplasmic reticulum of neurons. Inhibition of AChE activity was severe in groups treated with relatively low doses of malathion. Nicotinamide (5.0 mg/0.05 ml) alleviated malathion-induced morphological alterations. In conclusion, it is suggested that malathion changes the ultrastructure and reduces. AChE activity in differentiating neurons, and the severity of which is consistently dose- and age-dependent.

• 한국동물학회지
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• 제8권2호
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• pp.37-41
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• 1965

The frequency and pattern of sex chromatin in primary cultured cells of kidney cortex of cats and guinea-pigs, and muscle of chick embroys were examined and compared to those of in vivo condition, with special reference to the various cultured stages. 1. In cat, the frequencies of sex chromatin positive I of peripheral position were average 62.7% in female, and 15% in male, whereas those of non-peripherla position were 5.8% in female and 0.1% in male. The incident proportion between them showed a marked difference-approximately 10 times higher in female than male. These results failry indicated that a distinct nuclear dimorphism with regard to the sex chromatin positive I was established in cultured cells. The position of sex chormatin was usually peripheral location. The tendency of frequencies , with reference to the cultured stages, was low count in primary extracted and initial stage cells , but it showed a peaked frequency in 10-13 days after primary culture, and after that the frequencies were decreased gradually. Compared between I vitro and in vivo condition of the same tissues, the cells in vivo exhibited the sex chromatin in high frequency at the peak showed stage. 2. In guinea-pig , the frequencyies of peripheral positive I were 36.8% in female and 6.3% in male, while non-peripheral positions were 6.1% in female and 3.5% inmale. Its incident was a rate of nearly 4 times higher in female than male. The nuclear dimorphism was also established in cultrued cells of guinea-pig. The position and the incident frquency showed a similar pattern as in cat except the primary extracted cells. 3. In chick embryo, the frequencies of sex chromatin positive I of peripheral position were 38.2% in female, and 18.3% in male, non-peripheral position, however, was hardly to find. These results suggest that the definite sexual dimorphism was unable to find in chick embryo cultured cells. The position and the incident tendency were a similar pattern as in above mammals and the frequency was higher in vitro cells.

• 한국동물학회지
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• 제37권4호
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• pp.531-537
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• 1994

Partial internal amino acid sequences of the 15S ATPase from chick skeletal muscle were determined and found to be identical to the corresponding regions of the Mg2+_ATPase from Xenopus laevis oocytes, that is a close homolog of N-ethvlmaleimide-sensitive factor (called NSF) in hamster and Sec18p in yeast, both of which are believed to plaN an essential role in vesicle fusion in secretory process. Thus, the 15S Arpase in chick skeletal muscle maw also belong to a protein family of the "vesicle fusion proteins". Unlike the Mg2'-Afpase with an isoelectric point (pl) of 5.5, however, the 15S Arpase was separated into four spots with pls of 4.9,6.4 and 6.9 upon analysis by twoiimensional gel electrophoresis. In addition, the anti-15S ATPase IgG was found to be capable of interacting with the 265 protease complex upon analysis by immunoprecipitation. Moreover, immunoblot analysis revealed that the anti-155 Arpase IgG recognizes three subunits, ko of which show the same mobilities as the 510-kDa subunit 4 and 48-kDa subunit 7 of the 26S protease complex that are known to contain a highly consented ATP-binding motif. These results surest that a common antigenic site, likely the consensus nucleotide-binding site, exists in the 15S ATPase and the 26S pretense complex and hence both the enzymes maw also be related ATPases.d ATPases.

• 한국동물학회지
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• 제32권1호
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• pp.40-47
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• 1989

계배 근원세포의 배양 배지에 calcium ionophore A23187이나 EGTA를 배양 24시간에 첨가함으로서 초래된 세포내 칼슘 농도의 변화는 근원세포의 분화과정에 상당한 영향을 미쳤다. 배양 24시간에 A23187이나 EGTA를 첨가한 후 배양 48시간, 72시간, 및 96시간에 각각 세포를 [35S]methionine으로 1시간 표지시킨 후 수확하여 2차원 전기영동법으로 단백질을 분리시켰을 때, 일부 단백질은 배양 조건에 따라 합성 양상을 달리함을 보였다. 배양 24시간에 처리한 A23187과 calcium-activated neutral protease는 대조군에 비해 세포융합을 촉진시켰으나 동일 시기에 처리된 phosphoprotein을 정량함으로써 조사하였을 때, A23187이 배양 초기에는 대조군에 비해 약간 이 효소의 활성도를 높이는 효과를 보였으나 세포융합이 완성된 시기인 96시간에는 대조군에 비해 활성도를 높이는 효과를 보였으나 세포융합이 완성된 시기인 96시간에는 대조군에 비해 활성도의 차이를 나타내지 않았다. A23187 및 calcium-activated neutral protease에 의한 세포융합의 촉진, 그리고 A23187에 의한 protein kinase C 활성도의 증가가 모두 근원세포의 융합이 활발히 진행되는 시기인 배양 48-72 시간에 관찰됨을 볼 때, 세포내 칼슘의 농도는 protein kinase C 및 calcium-activated neutral protease와 상호연관을 가지면서 세포분화에 관여하는 것으로 사료된다.