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Immunogold Labellings and Expression of Metallothionein in Regenerating Rat Liver
안영모,오승한,김호진,이미영,이종화,신길상,김완종,Ahn, Young-Mo,Oh, Seung-Han,Kim, Ho-Jin,Lee, Mi-Young,Lee, Jong-Hwa,Shin, Kil-Sang,Kim, Wan-Jong Korean Society of Electron Microscopy 2005 Applied microscopy Vol.35 No.1
진핵세포 내에 존재하는 metallothionein (MT)은 시스테인 함량이 높은 저분자량의 단백질로서 2가 금속이온들과의 친화력이 높아 중금속에 대한 해독작용, 금속이온의 대사 및 세포분열 등과 관련되어 있다. 본 연구자들은 간 재생능력이 우수한 흰쥐를 실험모델로 하여, 간의 70% 정도를 실험적으로 제거한 후, 간 재생을 유도하는 과정에서 시간 경과에 따라 간세포의 미세구조 변화와 더불어 MT 유전자 발현 및 이 단백질의 세포내 분포를 알아보고자 하였다. 부분 간 절제 후 간 조직이 증식되고 재구성되어 간이 원래의 크기로 재생되는 시간은 1주일 정도가 소요되었다. 재생중인 간세포는 핵대 세포질의 비가 크고, 핵내에 인이 크게 발달하고 크기가 작은 미토콘드리아가 다수 나타났으며, 조면소포체가 잘 발달하고 있었다. MT mRNA는 간 절제 직후부터 증가하기 시작하여 1시간 경과 후에 최대치에 이르렀고, 12시간 이후부터 감소하기 시작하였다. 재생중인 간세포에서 MT에 대한 면역반응은 간 절제 후 12시간이 경과한 군에서 가장 높게 나타났고, 이후 점차 감소하여 8일이 경과한 실험군에서는 정상 대조군과 유사한 정도로 감소하는 것으로 관찰되었다. 또한, 간 재생 초기에는 항-MT 금 입자들이 주로 세포질쪽에 분포하다가 점차 핵내에 존재하는 양이 증가하는 것으로 나타났다. 이러한 결과들은 간 절제 후 보상작용으로 일어나는 세포분열 단계에서 MT가 이 과정에 필요한 요소를 제공하는 역할을 수행하기 때문인 것으로 사료된다. Metallothionein (MT) is a low molecular weight, cysteine-rich, metal-binding, and non-enzymatic protein. The present study was carried out to investigate the expression of MT gene as well as the localization of MT in regenerating rat liver. In partial hepatectomized rats, MT mRNA was detected as early as 1 hr and reached a maximal level by 8 hr after the operation. Thereafter, this level decreased gradually until 24 hr, and it became similar to that of sham control. Meanwhile, time course of MT immunoreactivity using immunogold-labelling revealed that the number of gold particles in hepatocytes increased significantly by 12 hr, but decreased at 48 hr after partial hepatectomy. Ultrastructurally, immunogold particles indicating the presence of MT were distributed in both the cytoplasm and the nucleus of the rat hepatocytes, particularly the particles were distributed at rough endoplasmic reticulum and nucleolus and did not seem to adhere to mitochondria or lysosomes in proliferating hepatocytes. Briefly, high level of MT mRNA expression and the intense immunoreactivity and/or the specific localization of MT was observed during liver regeneration. These results suggest that MT possibly involves hepatocyte proliferation via the storage or the supply of various metal ions in the regenerating rat liver.
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발생중인 흰쥐 간에서의 Metallothionein 발현에 관한 면역세포화학적 연구
오승한,안영모,신길상,김완종,Oh, Seung-Han,Ahn, Young-Mo,Shin, Kil-Sang,Kim, Wan-Jong 한국현미경학회 2004 Applied microscopy Vol.34 No.3
메탈로사이오닌 (metallothionein: MT)은 시스테인(cysteine)이 다수 포함되어 있고, 저분자량인 단백질로서 중금속이온이나 다양한 세포 독성 인자에 의해 유도되며, 최근 발암과정 (carcinogenesis)이나 혹은 세포분화와 연관하여 활발하게 연구가 이루어지고 있다. 본 연구는 흰쥐를 수정시킨 후 발생과정 또는 출생 후 성장과정 단계에서 태아와 신생 흰쥐의 간 조직 및 세포내 MT의 분포 양상을 조사하고자 하였다. 흰쥐 간 조직내 MT의 분포는 면역조직화학적 방법을 적용하여 광학현미경으로 관찰하였고, 세포내 미세구조적 위치 (localization)는 금 입자 표지를 이용한 면역세포화학적 방법을 이용하여 투과전자현미경으로 관찰하였다. MT는 발생 13일이 경과한 흰쥐 태아의 간에서 처음으로 관찰되기 시작하여, 출생 당일의 신생 흰쥐에서 가장 높게 관찰되었다. 면역 조직화학 및 세포화학적 방법으로 관찰한 결과, MT 염색성은 발생 후기에서 출생후 10일째까지 비교적 높게 나타났다. MT에 대한 양성반응은 난형 세포 (oval cell)들과 간 실질세포 (liver parenchymal cell)들에서 뚜렷하게 존재하였다. 발생과정과 신생 흰쥐의 간 조직에서는 핵과 세포질 양쪽에서 공통적으로 양성 반응을 보였으나, 생후 30일 이후부터 성체에 이르는 시기까지는 세포질에 한정되어 있는 양상을 보였다. 금 입자 표지법에 의한 MT의 세포질내 분포를 보면, 발생 중인 흰쥐 간 세포에서는 핵과 세포질내에 고르게 산재되어 있었다. 즉, 핵내 염색질, 인, 조면소포체 및 cytosol에서도 MT에 대한 면역 금 입자들이 관찰되었으나, 미토콘드리아와 지질 소포 등에서는 거의 관찰되지 않았다. 상기의 결과들은 발생중 혹은 성장중인 흰쥐의 간에서 MT가 점차 증가하는 것으로 요약될 수 있는 것으로서, 이는 MT가 세포들의 증식과 분화에 금속 이온의 저장과 방출 혹은 전사인자 (transcription factor)로서 작용할 수 있음을 암시하는 것으로 판단된다. Metallothionein (MT) is a family of ubiquitous, low molecular weight (6-7 kDa), cysteine-rich protein with a high affinity to metal ions and has no aromatic amino acids and histidine. Some of the known functions of MT include detoxification of heavy metals and alkylating agents and neutralization of free radicals. Also, this protein may affect a number of cellular processes including gene expression, apoptosis, proliferation and differentiation. But, its actual functions are still not clear. The present study was undertaken to examine immunocytochemically the localization of MT in developing rat liver. On the day 11 of gestation, the fetal rat liver has already been formed and contained numerous oval cells with high nuclear cytoplasmic ratio, which were the progenitors of hepatic parenchymal cells, but no reaction products of MT were detected at this time. And then, positive reactions against MT started to appear predominantly in the parenchymal cells of liver from the 13th day after gestation. Reaction products, immunogold particles or brown coloration, were localized at both the nucleus and the cytoplasm of the parenchymal cells, except mitochondria. The intensity of this reaction gradually increased, and exhibited the strongest at birth. The intensity of MT staining and immunogold labelling diminished with growth, and by the 15th day after birth weak positive reaction was observed in the cells. In brief, positive reactions for MT were observed in the oval cells and the parenchymal cells during fetal stage, meanwhile they were present only in the parenchymal cells after birth. The present results suggest that MT possibly involves parechymal cell proliferation and differentiation through the storage or the supply of various metal ions in the developing rat liver.
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박영성(Yeong-Seong Park),김경수(Kyung-Soo Kim),안영모(Young-Mo Ahn),최선미(Sun-Mi Choi),최욱(Uk Choi) 대전대학교 환경문제연구소 2008 환경문제연구소 논문집 Vol.12 No.-
Characteristics of VOC(benzene) treatment were investigated using a fixed bed reactor system over copper base catalyst and photocatalyst/adsorbent blending material. The copper base catalysts were made by using copper nitrate reagent and various support materials such as γ-Al₂O₃, TiO₂. The parametric tests were conducted at the reaction temperature range of 200~400℃, benzene concentration of1,000~2,000 ppm, and space velocity range of 5,000~10,000 hr<sup>-1</sup>. The property analyses such as BET, SEM and the removal efficiency(conversion) of VOC were examined. The experimental results showed that the VOC removal efficiency of hybrid method was higher than that of single method. It was also found that the comprehensive feasibility study of hybrid method would need with considering various factors including additional expenses.
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안영모,김완종 순천향대학교 기초과학연구소 2004 순천향자연과학연구 논문집 Vol.10 No.2
Metallothionein(MT) is a low molecular weight (approximately 6,000~7,000Da), cysteine-rich protein (approximately 22~23%), metal-binding protein. MT is known to be induced by surgical operation and chemical agent, as carbon tetrachloride(CCI_(4)) and cadmium(Cd). The goal of the present study was to determine the distribution of the protein on regenerating rat liver after partial hepatectomy(PH). The results were as follow: Size of the remained liver increased at 1day after PH. Morphological changes were observed such as destroyed hepatocyte plate around central vein, enlarged nuclear cytoplasmic ratio and increased the number of hepatocytes with two nuclei. Expression of MT mRNA started to increase at 30 minutes, reached to maximum at 1 hour and maintained at 8 hours after PH. The level of MT mRNA showed similar to that of control group at 12 hours. On immunohistochemistry, in all groups treated with primary antibody, immunoreactivity indicating the presence of MT appeared moderately throughout both the cytoplasm and nucleus of rat hepatocyte. On the immunohistochemical analysis, immunoreactivities both in the cytoplasm and nucleus appeared after 4 hours, reached to maximum at 12 hours, and then it showed similar to those of normal control. These results suggested that the remained liver immediately entered cell proliferation and this MT protein during the process may be associated with transfer of factors needed to cell reproduction from the cytoplasm to the nucleus for regeneration of the liver after PH.
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안영모,김완종,신길상 순천향대학교 기초과학연구소 2001 순천향자연과학연구 논문집 Vol.7 No.1
Ultrastructural changes during spermatogenesis in the hamster testis was studied by electron microscope. The purpose of the present study is to ascertain the ultrastructural changes of various germ cells, Sertoli and Leydig cells. The results were as follows: various staged germ cells were embedded in cytoplasm of Sertoli cell. Leydig cell contained numerous mitochondria with tubular cristae and well developed smooth endoplasmic reticulum. Spermatogonia were located close to the basal layer. They were spherical to somewhat oval cells with large nucleus. Spermatocytes were connected to each other until the late spermatid stage by intercellular bridge. An acrosome was located apical head and microtubule arrangement of flagellum was shown typical "9+2" characteristics. In conclusion, spermatogenesis involves the development, separation and maturation of germ cells, and Sertoli and Leydig cells show similar ultrastructures compared with ones of other mammalian species.
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