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      • 계배심근 근원섬유형성에 관한 면역형장, 면역전자현미경 및 전기영동 연구

        하재청,김동수 한국통합생물학회 1992 동물학회지 Vol.35 No.3

        심근세포는 골격근 세포와 더불어 세포의 분화기작을 이해하기 위한 좋은 모델이라 할 수 있다. 본 연구에서는 심근세포의 배양중에 일어나는 심근 근원섬유형성과정에 대한 이해를 위하여 SDS- 및 two-dimensional gel electrophoresis와 immunofluorescent 및 immunoelectron microscopy를 수단으로 이를 검토하였다. 배양과정에서 나타나는 주요한 단백질 중에서 SDS-PAGE에 의해 209 kDa와 53 kDa 및 46 kDa band가 각각 막분획 및 세포질 분획 단백질에서 배양과정동안에 가장 현저한 양상을 보였다. 특히 배양 3일과 4일의 양상에서 뚜렷한 변화를 나타내어 이들의 OD 격은 배양 96시간에서의 막분획에서 0.38(209 kDa)와 0.52(53 kDa) 및 배양 72시간에서의 세포질 분획에서 0.34(46 kDa)로 각각 최고치를 나타내었다. 또한 이차원 전기영동상의 양상에서도 세포질 및 막분획 단백질의 전반적인 전개양상이 배양 96시간에서 가장 두드러짐으로써 결국 배양 72시간 및 96시간대에서 심근 모세포의 분화와 관련된 일련의 단백질 합성과정이 가장 활발하다는 것을 알 수 있었다. 배양중에 발현되는 actin에 대한 형광현미경적 분석에 의해, actin은 세포내에서 불균등하게 분포하였고 근원섬유가 형성되는 시기인 배양 96시간에 세포외곽부에서 나타나는 다소 강한 염색성을 관찰할 수 있었다. 심근세포의 근원섬유는 전자현미경 관찰에 의해 primitive forms로 배양 96시간에 출현함을 알 수 있었으며 이 시기의 근원섬유의 형태는 여러 구성대(A, H and M bands)가 미약하나 1대는 비교적 뚜렷하였다. 따라서 심근세포의 T근원섬유형성과정에서 관찰되는 뚜렷한 단백질양상의 변화와 근원섬유의 출현시기 및 구성과정간에는 중요한 관련성이 있으며 이러한 과정에서 actin은 세포질내에서 불균등한 분포를 나타낸다는 것을 알 수 있었다.

      • KCI등재

        계배 흉근 근섬유 형성에 대한 염화수은의 영향

        하재청,김한도,김병기,김정락,강호성,최은상,Hah, Jae-Chung,Kim, Han-Do,Kim, Byeong-Gee,Kim, Chong-Rak,Kang, Ho-Sung,Choe, Eun-Sang 한국현미경학회 1993 Applied microscopy Vol.23 No.2

        To investigate the effect of mercuric chloride on the pectoral muscel cells during chick embryogensis, chicken embryos were treated with mercuric chloride. Monoclonal antibodies against myosin were prepared for the localization of differentiation of thick filaments detected by the use of protein A-gold complex as a cytochemical marker. The effect of mercuric chloride was appeared not only to be ascribed with the reduced formation of myofibrilogenesis but also associated with induced change of morphology by the inhibition of protein synthesis.

      • The Effects of Mercuric Chloride on the Protein Synthesis during Myogenesis in the Chicken Breast Muscle

        하재청,김병기,김한도,강호성,김정락,Hah, Jae-Chung,Kim, Byeong-Gee,Kim, Han-Do,Kang, Ho-Sung,Kim, Chong-Rak 생화학분자생물학회 1993 한국생화학회지 Vol.26 No.1

        11일 된 계배의 흉근에서 얻어낸 myoblast의 배양에서, 5ppm 농도의 수은은 세포성장과 분화과정중 형태척인 변화에는 별다른 영향을 끼치지 않았다. 또한 myoblast에서 myotube로 분화되는 과정에 일어나는 세포융합은 수은에 의하여 미미하게 억제되었으나 단백질합성에는 상당한 변화를 초래하였다. Cytosolic soluble protein으로 판독된 4종류의 단백질은 모두다 세포성장분화와 더불어 합성이 증가되었다. 특히 Western blot에서 actin으로 나타난 46kDa 단백질이 myotube가 많이 형성되는 4일째 배양세포에서 두드러졌다. 그러나 수은처리군에서는 46kDa 단백질을 위시한 모든 단백질의 세포내 농도가 격감하였다. 한편, membrane-associated protein으로서는 7종류의 판독된 단백질중 200 kDa 단백질의 변화가 주목되었는데, 이 단백질은 standard myosin heavy chain과 comigration 하였으며 Western blotting에 의하여 myosin으로 나타났다. 그리고 세포분화 양상이 나타나면서 MHC 단백질합성이 증대되었으나 수은 처리군에서는 이 단백질은 검출되지 않았다. 위의 결과에 의하면 myoblast의 분화과정에서 필수적인 세포융합은 근수축단백질인 actin과 myosin이 중요한 역할을 하는데 이들 단백질은 세포질내에 산재해 있기 보다는 세포막과 밀접한 관계를 갖고 막주위에 위치한 것으로 추정된다. 그리고 비록 5ppm의 수은이 4일된 배양세포에서 극히 미미한 범위 내에서 세포분화를 억제하였지만 근수축단백질을 위시한 여러 단백질의 합성을 명확히 억제하였다. Mercuric chloride at the concentration of 5 ppm did not cause any significant morphological alteration except slightly low fusion index in the differentiation of myoblasts of chicken breast muscle tissue. However, it gave some influences in the patterns of protein synthesis during the formation of myotubes. The concentrations of all four major proteins in the cytosolic fraction increased along with the culture age. Among the proteins, 46 kDa protein, which was actin, was the most prominent one and sharply increased as the myotube formation was apparent. However, the increase of protein could not be detected in the culture treated with $HgCl_2$. In the membrane fraction, protein of 200 kDa, which was myosin, only showed an increase during myogenesis. However, myosin was not found in the presence of mercury, could not be detected in untreated cytosolic fraction, suggesting peripheral distribution of myosin during the cell fusion in the early stage of myogenesis. The increases of myosin in membrane fraction and actin in cytosolic fraction indicate that contractile proteins are presumable associated with cell membrane, and that membrane fusion is an important event during myogenesis.

      • An Experimental Study on the metabolism of 5-Hydroxytryptamine

        하재청,이원호,Hah, Jae Chung,Lee, Wuen Ho The Korean Society for Integrative Biology 1973 동물학회지 Vol.16 No.1

        The changes in the quantitative distribution and in cytoplasmic granules of tongue mast cells and duodenal enterochromaffin cells in male albino rats were observed following oral administration of 40mg/kg body wt. isonicotinic acid hydraside (INH) and 20mg/kg body wt. pyridoxine. The results obtained are summarized as follows: 1. INH administered-rat showed a marked decrease in the number of mast cells, caused by leakage of cytoplasmic granules, while pyridoxine-rat showed increased the number of mast cells. 2. Similarly, INH-rat showed a marked decrease in the number of enterochromaffin cells. In the case of pyridoxine-rat, however, the number of enterochromaffin cells increased compared with that of the controls. 3. In view of the fact that a large dose of INH was harmful to the formation of mast cells and enterochromaffin cells. And considering that a moderate dose of pyridoxine stimulated the formation of the two kinds of cells and the amounts of cytoplasmic granules, it was concluded that pyridoxine might be concerned with the metabolism of secretory products, 5-Hydroxytryptamine.

      • Histochemistry of the Developing Chick Respiratory Tract

        하재청,Hah, Jae Chung The Korean Society for Integrative Biology 1972 동물학회지 Vol.15 No.4

        A histochemical study of the respiratory tract in developing chick was done to demonstrate PAS-postivie materials, ribonucleic acid, phospholipid, and alkaline phosphatase activity. Following results are obtained: 1. The alkaline phosphatase activity was found to be high before the appearance of cartilage in mesenchyme surrounding the tracheal epithelium. The enzyme activity declined after the cartilage formation, followed by the restricted activity in epithelium in the postembryonic stage. 2. A moderate positive reaction of ribonucleic acid was found in the cytoplasm of the epithelium and undifferentiated mesenchyme. As the cartilage grew differentiated the reaction of ribonucleic acid was found to disappear in the mesenchyme surrounding the epithelim, but the cytoplasm of the glands showed a moderate positive reaction. 3. Goblet cells of the mucosa and glandular cells showed highly positive reaction, but the basement membrane exhibited slightly positive PAS-reaction. 4. Epithelial cells of the mucosa showed a weak to moderate reaction. However, the epithelia of bronchiol and alveoli in the differentiating period and glandular cells showed a strong positive reaction in Baker's hematein test.

      • SCIESCOPUSKCI등재

        계배 흉근세포의 Myogenesis 에 있어서 염화수은이 단백질합성에 미치는 영향

        하재청,김병기,김한도,강호성,김정락 ( Jae Chung Hah,Byeong Gee Kim,Han Do Kim,Ho Sung Kang,Chong Rak Kim ) 생화학분자생물학회 1993 BMB Reports Vol.26 No.1

        Mercuric chloride at the concentration of 5 ppm did not cause any significant morphological alteration except slightly low fusion index in the differentiation of myoblasts of chicken breast muscle tissue. However, it gave some influences in the patterns of protein synthesis during the formation of myotubes. The concentrations of all four major proteins in the cytosolic fraction increased along with the culture age. Among the proteins, 46 kDa protein, which was actin, was the most prominent one and sharply increased as the myotube formation was apparent. However, the increase of protein could not be detected in the culture treated with HgCl₂. In the membrane fraction, protein of 200 kDa, which was myosin, only showed an increase during myogenesis. However, myosin was not found in the presence of mercury, could not be detected in untreated cytosolic fraction, suggesting peripheral distribution of myosin during the cell fusion in the early stage of myogenesis. The increases of myosin in membrane fraction and actin in cytosolic fraction indicate that contractile proteins are presumable associated with cell membrane, and that membrane fusion is an important event during myogenesis.

      • Distribution of Alkaline Phosphatase, PAS-Positive Materials and Phospholipid in the Kidney of the Developing Chick

        하재청,Hah, Jae Chung The Korean Society for Integrative Biology 1969 동물학회지 Vol.12 No.3

        著者는 白色레그혼의 種卵을 孵化시켜 얻은 鷄胎中 賢과 後賢에 있어서의 alk. p., PAS-陽性物質 및 燐脂質의 消長과 局在位置의 變異를 組織化學的 方法으로 追究하여 다음과 같은 結果를 얻었다. 1. 分泌尿細管의 器官形成에 앞서 未分化 間葉組織內에 强한 alk. p. 活性이 出現하였으며 同活性은 分泌尿細管의 分化와 함께 消失하였다. 2. 中賢 및 後賢 分泌尿細管의 分化와 함께 alk. p. 및 PAS-陽性物質은 分泌尿細管의 刷子綠에 强陽性反應을 보였다. 3. 中賢 및 後賢 分泌尿細管의 細胞質과 刷子綠은 强한 燐脂質 陽性反應을 띠었다. 4. 造賢組織의 分化中 未分化 間葉組織에 alk. p. 活性이 出現하였다가 組織의 分化와 함께 消失된 事實에서 보아 alk. p. 는 組織分化에 關與하는 것으로 생각된다. 5. 中賢 및 後賢 分泌尿細管의 刷子 에 있어서 alk. p. 와 PAS-陽性物質 및 燐脂質의 變異와 分布狀態가 一致한 것으로 보아 alk. p. 는 炭水化物의 再吸收 및 燐脂質代謝에 關與하는 것으로 思料된다. For the purpose of making clear the activity of the alkaline phosphatase to the morphogenesis and function of the various structures of the developing chick kidney in relation to PAS-positive materials and phospholipid, the author observed histochemically the aforementioned enzyme and other substances. The mesonephros and metanephros of 4-20 day white leghorn embryos were used, obtaining the following results. 1. Before definite appearance of the secretory tubules the alkaline phosphatase activity showed strongly in the undifferentiated mesenchymal tissue. As the tubules grew differentiated, the alkaline phosphatase activity was found to have disappeared in the mesenchymal tissue surrounding the tubules. The above mentioned fact indicates that the alkaline phosphatase may be concerned with morphogenesis of the developing chick kindney. 2. The fact that the strong alkaline phosphatase activity and the occurrence of the PAS-positive materials were observed at the luminal borders of differentiated secretory tubules of mesonephros and metanephros, indicates that alkaline phosphatase may be concerned with reabsorption of carbohydrate at the borders. 3. A strong positive reaction of phospholipid was found in the cytoplasm and brush borders of the mesonephric and metanephric tubules. The fact that vicissitude of alkaline phosphatase was found to coincide with that of phospholipid suggests that the enzyme may have influence on the metabolism of the phospholipid.

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