재생중인 흰쥐 간세포의 조직화학적 미세구조 관찰

최치용,손성향,유창규,최임순,Choi, Chee-Yong,Sohn, Seong-Hyang,Yoo, Chang-Kyu,Choe, Rim-Soon 한국현미경학회 1988 Applied microscopy Vol.18 No.2

An ultrastructural study of hepatocyte proliferation in the regenerating rat liver has been made by means of the partial hepatectomy. And electron microscopic histochemistry of hepatocyte in the regenerating rat liver is studied through alkaline phosphatase reaction. The results are as follows: 1. When the regeneration of rat liver is induced by the partial hepatectomy, the prominent ultrastructural characteristics of hepatocyte are changes of the distribution of chromatin in nucleus, increase of the number of mitochondria and decrease of the size of them, development of rough endoplasmic reticulum, and transient decrease of glycogen granules in cytoplasm. 2. Alkaline phosphatase reaction products are appeared in the nucleus or rough endoplasmic reticulum of hepatocyte during the initial regeneration of liver as 24, 48 and 72 hour groups after partial hepatectomy. And these positive reaction are mainly increased in cytoplasm and plasma membrane of hepatocytes during 1, 2 and 3 week groups after partial hepatectomy. As 4 weeks passed after partial hepatectomy, these positive reaction is located in the sinusoidal epithelial cells or erythrocytes. With above results, we concluded that alkaline phosphatase was synthesized in the rough endoplasmic reticulum bounded ribosomes of regenerating hepatocyte, was transported to the plasma membrane of them, and then was transported in blood by the way sinusoidel epithelial cells.

흰쥐 간세포의 소포체에서의 Aflatoxin B1 의 대사 및 돌연변이 유발양상에 미치는 Butylated Hydroxyanisole 의 영향

최치용,차영남 ( Chee Yong Choi,Young Nam Cha ) 한국환경생물학회 1991 환경생물 : 환경생물학회지 Vol.9 No.1

Effect of administering butylated hydroxyanisole(BHA) on the metabolism of aflatoxin B_1(AFB_1) and production of mutagenic metabolites have been compared with those of phenobarbital(PB) and 3-methylcholanthrene(MC) administration in rat liver microsomes. Male Sprague-Dawley rats were treated with these inducers and liver microsomes were isolated. These microsomes were used to metabolize AFB_1 and to produce mutagenic metabolites. Results showed that normal rat liver were able to metabolize AFB_1 quite actively and produced large amounts of AFB-8, 9-epoxide (appearing as the AFB-8, 9-dihydrodiol-Tris complex). Upon incubations of normal rat liver microsomes with increasing concentrations of AFB_1, a steep dose-related increases of mutagenicity were observed in the Ames test. The PB-microsomes had an increased ability to metabolize AFB_1 and particularly the rate for the production of the weakly mutagenic AFQ_1 metabolite was markedly increased. Conversely, PB-microsomes had a moderate decrease in its ability to form the strongly mutagenic of AFB-8, 9-epoxide metabolite. However, the ability of PB-microsomes to form mutagenic metabolites from AFB_1 was somewhat greater than that of the control-microsomes. The MC-microsomes had an increased ability to metabolize AFB_1 also. However, instead of the weakly mutagenic AFQ_1 metabolite seen with the PB-microsomes, large amounts of the strongly mutagenic AFM_1 metabolite was formed. Although AFM_1 is not known to be a direct mutagen, it was highly mutagenic upon activation with microsomes. The very steep dose-related increases of mutagenicity and appearance of bacterial toxicity at relatively lower doses of AFB_1 may have been caused by the secondary metabolic activation. The ability of BHA-microsomes to metabolize AFB_1 was decreased. Among the metabolites produced by the BHA-microsomes, the non-mutagenic AFB_2a, was formed in significantly increased amounts but the toxic AFB-8, 9-epoxide was produced only in much reduced amounts. The AFB_2a was not mutagenic even after metabolic activation with microsomes. When increasing concentrations of AFB_1 was incubated with BHA-microsomes, a very mild dose-related increases of mutagenicity was observed and the occurence of toxic effects on bacterial growth appeared only at high doses of AFB_1, This may have been due both to the reduced rate of overall AFB_1 metabolism and to the decreased formation of the highly mutagenic AFB-8, 9-epoxide but an increased formation of the non-mutagenic AFB_2a metabolite by the BHA-microsomes. Such a reduced rate of formation of the toxic AFB-8, 9-epoxide but an increased production of the non-toxic AFB_2a metabolite by the BHA-microsomes may have been, at least partially, responsible for the anticarcinogenic effect of BHA.

Aflatoxin $B_1$으로 유발되는 흰쥐 간세포의 미세구조 변화 : Butylated hydroxyanisole(BHA) 전처리에 의한 보호 효과

최치용,최임순,차영남,Choi, Chee-Yong,Choe, Rim-Soon,Cha, Young-Nam 한국현미경학회 1991 Applied microscopy Vol.21 No.1

Butylated hrdroxyanisole(BHA), a widely used food additive phenolic antioxidant, is known to inhibit cancer formations inducible with a wide variety of chemical carcinogens including aflatoxin $B_1(AFB_1)$. Thus, in the present study morphological characteristics underlying the hepatoprotective effects of BHA against $AFB_1$ inducible ultrastructural changes of hepatocytes have been examined. The obtained results are as follows : 1 . Livers obtained from rats treated with $AFB_1$ in vivo have been examined with transmission electron microscope. Among the many hepatocellular structural aberrations induced by $AFB_1$ treatment, the nuclear chromatins were found to be distributed irregularly('cap formation') and the nuclear membrane was found to be partially segregated. Furthermore, there were many lipid droplets, hyperplasia of smooth endoplasmic reticulum, dialated rough endoplasmic reticulum and, lysosomes arrested at various stages of its development. 2. Also, when $AFB_1$ was given in vitro to hepatocytes which have been isolated from untreated normal rats and examined under scanning electron microscope, there were much 'blobbing' phenomena resulting from cytoskeletal disturbances. 3. However, in the liver obtained from rats pretreated with BHA and then give the $AFB_1$, the observed morphological aberrations were in much reduced extent. Similarly, the BHA-hepatocytes had much decreased severity in the $AFB_1$ inducible blob formations.

흰쥐 간세포의 소포체에서의 Aflatoxin B1 의 대사 및 돌연변이 유발양상에 미치는 Butylated Hydroxyanisole 의 영향

최치용,차영남 한국환경생물학회 1991 환경생물 : 환경생물학회지 Vol.9 No.1

Actinomycin D가 흰쥐 원생식세포 이동에 미치는 영향에 관한 조직화학적 미세구조 관찰

최치용,최춘근 연세대학교 자연과학연구소 1984 學術論文集 Vol.13 No.-

본 연구에서는 임신한 쥐에 Actinomycin D를 투여하여 원생식세포의 분포로써 이동경로와 시간을 조사하였으며, 각 이동경로에 따르는 형태학적 차이와 미세구조의 변화를 alkaline phosphatase로 조직화학적 방법으로 처리하여 전자현미경으로 관찰하였다. 그 결과 흰쥐 발생과정에서 원생식세포는 수태 9일째 난황낭의 내배엽 세포에서 유래하여 수태 11일째 후장을 지나며 수태 13일째에는 배측장간막을 경유하며 수태 14일째 생식융기에 도달하였다. 또한 원생식세포가 후장과 생식융기에 있을때 alkaline phosphatase 에 대한 양성반응이 크게 나타났으며, Actinomycin D를 투여하였을 때에는 이에 대한 반응이 현저히 줄어들었다. 원생식세포의 형태학적 변화로 pseudopodia 와 tail process는 후장과 배측장간막을 지날때에 가장 잘 나타났으며, 이때에는 trailing cytoplasm도 잘 볼 수 있다. 그리고 특수한 세포소기관인 nuage는 발생 초에는 나타나지 않고 이동함에 따라서 많이 나타났다. 본 실험의 결과를 통하여 원생식세포의 이동은 세포형태의 변형에 의하여 진행되며, 이러한 특징이 Actinomycin D에 의하여 손상을 입었을 때에 원생식세포의 이동이 억제되었다. 따라서 이와 같은 현상이 원생식세포가 이동할 수 있는 형태학적 특징임을 확인할 수 있다. In this study, pregnant rats were injected with Actinomycin D and the reaction of Alkaline phosphatase was used for identifying the Primordial germ cells (PGCs). So the pathway and date of migrating PGCs were observed light microscopically as the distribution of PGCs, and ultrastructural changes were observed electron microscopically during migration of PGCs. At the 9th gestation day, PGCs were observed among the endodermal cells of yolk sac, at the 11th gestation day, PGCs were seen in the hindgut and then entered the dorsal mesentery by the 13th gestation day, and at the 14th gestation day, the genital ridges were populated with them. When PGCs located in the hindgut and genital ridges, the positive reactions of Alkaline phosphatase were dominated, in the case of Actinomycin D treatment these reaction were lessened. It appeared such as pseudopodia, tail process, trailing cytoplasm and nuage as the ultrastructural characteristics of PGCs. In addition, these morphological features were damaged by Actinomycin D treatment. In these results, it was convinced that migration of Primordial germ cells went through these morphological features.

흰쥐 부정소 상피세포의 여러 유형에 관한 연구

정경순,박용빈,최치용,고기석,최임순,Cheong, Kyung-Soon,Park, Yong-Bin,Choi, Chee-Yong,Koh, Ki-Seok,Choe, Rim-Soon 한국현미경학회 1990 Applied microscopy Vol.20 No.2

Several types of the epithelial cells were classified by ultrastructural observation through transmission electron microscope in the rat epididymis. Ultrastructural studies showed that the principal cells, basal cells and narrow cells are located in all the regions of the epididymis and the light cells are present only in the corpus and cauda epididymis. It was observed that the columnar epithelial cells like the principal cells, light cells and narrow cells contain several secretory vesicles and there are halo cells migrating in the several regions. The basal cells showed the elliptical forms in the caput and corpus region and the global forms in the cauda region.

Actinomycin D가 흰쥐의 모체 및 태아 간세포에 미치는 영향에 관한 전자현미경적 연구

한금자,고기석,최치용,최춘근,최임순,Hahn, K.J.,Ko, K.S.,Choi, C.Y.,Choi, C.K.,Choe, R.S. 한국현미경학회 1983 Applied microscopy Vol.13 No.1

This study was made to investigate the ultrastructural changes of the hepatocyte of the maternal liver, and fetal liver by Actinomycin D in Wistar rats at the stage of pregnancy. Peritoneal injection of Actinomycin D to rats carried out gestation day 7 to 9 at the level of $15{\mu}g(11.5{\mu}g/100g$ body wt.), $20{\mu}g(15.8{\mu}g/100g$ body wt.) on each day. Treated animals with saline only were used for controls. Animals were sacrificed on day 15 of gestation. On electron microscopic examination, the hepatocytes of maternal liver given Actinomycin D $15{\mu}g$/ml had evidence of serious cellular damage, for example, hypertrophy of rough endoplasmic reticulum, loss in nucleolar osmiophilia, swelling of Golgi apparatus and change of mitochondrial structure. Maternal liver given Actinomycin D $20{\mu}g/ml$ shown similar changes to that of the $15{\mu}g/ml$ treated animals. But mitochondria of this group were not changed than that of $15{\mu}g/ml$ treated group. In the hepatocytes of fetal liver, changes were more pronounced. The drug produced alteration in nuclei and cytoplasm. The rough endoplasmic reticulum was swollen and there were ribosomes detachement. In addition, damages of mitochondria, Golgi apparatus were detected.

간세포 미토콘드리아의 호흡에 미치는 비소의 영향

부문종,이강오,최임순,최치용 ( Moon Jong Boo,Kang Oh Lee,Rim Soon Choe,Chee Yong Choi ) 한국환경생물학회 1991 환경생물 : 환경생물학회지 Vol.9 No.2

To understand the effect of Arsenic(As) on the rat liver mitochondria, various amounts of As were added to the mitochondrial preparation. When pyruvate and malate were used as respiration substrate, the mitochondrial respiration was inhibited in the presence of 0.1∼1.0mM of As, but when succinate was used as substrate, the respiration was only inhibited at 1.0∼2.0mM of As. This suggests that As might inhibit NADH ubiquinone reductase(complex Ⅰ). 0.4 mM of As reduced the activity of complex Ⅰ and brought about induction of swelling and inhibition of ATP driven contraction of mitochondria. These experiments again suggest that As has an inhibitory effect on the mitochondrial respiration by selectively inhibiting the electron transport mediated by complex Ⅰ of respiratory chain and by affecting the integrity of mitochondrial membrane.

쥐 간세포의 노화에 따른 형태학 및 생화학적 연구

최임순,주충노,최춘근,최치용,고기석,고지훈,곽한식 연세대학교 자연과학연구소 1983 學術論文集 Vol.11 No.-

생후 3개월부터 25개월까지 성장한 쥐를 나이군에 따라서 간세포의 미세구조적 변화를 전자현미경으로 관찰하고, DNA,RNA 및 단백질의 함량변화와 몇가지 효소활성의 변화를 관찰하였다. 미세구조적 변화로서 나이가 증가함에 따라 mitochondria의 크기가 커지며 수가 감소하는 현상이 나타나고 Lipid droplet가 늙은군에서 많아졌다. 한편 DNA,RNA및 단백질 함량의 변화 관찰로부터 나이의 증가에 따라서 단백질합성의 효율이 저하됨을 알 수 있으며 SDH, LDH, G6-PDH 효소 활성의 측정 결과 어린군에서는 성장에 필요한 생합성을 위하여 효소활성이 높아진다는 것을 알 수 있었다. 그러나 이러한 효소활동은 나이의 증가에 따라 점차로 감소되는 경향을 보였고, 아주 늙은 쥐에서의 효소활성은 크게 저하됨을 알 수 있었다. 이와같은 실험결과로 부터 세포는 노화됨에 따라 대사 기능이 저하되고 아주 늙은 쥐에서는 mitochondria와 핵과 같은 세포소기관의 형태적 변화까지 초래되는 것으로 생각된다. The liver cells of normal rats of young (3 month and 6 month old), middle age (9 month, 12 month, and 15 month), old age (18 month, 20 month, and 23 month old) and very old age (25 month old) were investigated morphologically as well as biochemically. Observation of ultrastructure of the cells of normal rats using electron microscope showed that the number of the mitochondria was decreased as aging but their size became larger at their older age. Furthermore, the lipid droplet in the cytoplasm appeared frequently in the cells of older group. The DNA, RNA, and protein analysis showed that the efficiency of the protein biosynthesis became lowered as aging. The activities of succinate dehydrogenase, glucose 6-phosphate dehydrogenase and lactate dehydrogenase were also gradually lowerd as aging. From the above experimental results, it seemed likely that the cell activities became gradually lower and the physiological activities of the extremely old rats became extremely low and some morphological changes of organells such as mitochondria and nucleus occured.