붕어(Carassius carassius)의 조직내 젖산수소이탈효소와 에스테라아제 아이소자임에 미치는 동의 영향에 관한 연구

Studies on the Effects of Copper on the Lactate Dehydrogenase and Esterase Isozymes in Various Tissues of Carassius carassius

붕어(Carassius carassius)에 미치는 동의 영향을 밝히기 위하여 다음과 같은 연구를 하였다. 1) 셀루로오스 아세테이트 전기영동법에 의한 젖산수소이탈효소 아이소자임 상, 2) 분광비색법에 의한 LDH활성과 LDH효소계에 대한 동의 영향, 3) 한천 박층 전기영동법에 의한 에스테라아제 아이소자임 상, 4) 전분 겔 전기영동법에 의한 혈색소 상, 및 5) 조직 학적 연구. 1. LDH 아이소자임 영동대는 정상 붕어의 아가미에 2개 (LDH-3 및 LDH-5), 간에 3개(LDH-2, LDH-4, 및 LDH-5), 그리고 근육에서 2개 (LDH-3 및 LDH-4) 나타났다. LDH-1은 이상의 세가지 조직에서 나타나지 않았다. 동을 처리한 붕어의 간에서는 LDH-3이 나타났고 근육에서는 LDH-5가 나타났다. 그러나 아가미에서는 새로운 영동대가 나타나지 않았다. 2. LDH활성은 정상 붕어의 아가미, 간, 및 근육 중 아가미에서 가장 낯았고 근육에서 가장 높았으며, 아가미를 제외한 간과 근육에서는 1일부터 10일간의 동처리에 따라 점차적으로 감소되었다. 3. 동을 처리한 붕어의 간과 근육의 LDH활성 감소는 주로 체내 M-LDH에 대한 동의 억제에 기인된다. 4. 정상 붕어의 아가미, 간, 근육, 혈액, 뇌, 및 신장의 에스테라아제 아이소자임 영동대 수는 각각 3,6,2,2,2 및 2개 였고 이들은 동을 처리한 경우에서도 같았다. 동처리군의 아가미, 간, 혈액 그리고 신장의 에스테라아제 영동대의 상대이동도는 대조군의 그것들과 상이하였다. 5. 정상 붕어의 혈색소 영동대는 양극에 1개 있었다. 동을 처리한 붕어의 혈색소 영동대의 이동도는 정상 붕어의 그 것보다 약간 빠른 것으로 보였다. 6. 동(20ppm)에 5일간 처리한 붕어의 조직학적 연구는 다음과 같았다. 1) 아가미에서는 처음에 표피층이 분리되었다가 분해된 후 완전히 파괴되었다. 2) 간에서는 많은 지방립이 나타났다. 3) 정상 붕어와 동에 처리된 붕어의 근육 사이에서는 조직학적 차이가 없었다. 4) 신장족직은 기부요세관에 손상을 받았다. 요세관세포의 높이가 작아진 후 요세관의 내강이 확대되었다. 많은 기부요세관이 분홍빛의 과립성 형태와 여러 단계의 변성을 나타내었다.

In order to elucidate the effects of copper on Corassius carassius, the following were studied: 1) lactate dehydrogenase isozyme patterns by cellulose acetate electrophoresis, 2) LDH activity and copper effect on LDH enzyme system y spectrophotometry, 3) esterase isozyme patterns by agar thin layer electrophoresis, 4) hemoglobin patterns by starch gel electrophoresis, and 5) histological study. 1. There were two bands of LDH isozymes (LDH-3 and LDH-5) in the gill, three bands (LDH-2, LDH-4, and LDH-5) in the liver, and two bands (LDH-3 and LDH-4) in the muscle of the normal fish. The LDH-1 bond was not found in the above three tissues. When the fish were exposed to copper, LDH-3 appeared in the liver, LDH-5 in the muscle, but no new LDH band appeared in the gill. 2. The sepcific activities of the LDH were lowest in the gill and highest in the muscle of the normal fish, and they were gradually decreassed in the gill and highest in the muscle of the normal fish, and they were gradually decreased in the liver and mucle except in the gill from 1-day to 10-day exposure to copper. It indicates that LDH activities in the liver and muscle of the fish were inhibited by copper. 3. Through in vitro experiment, it is clear that the decrease of the LDH activities of the liver and muscle of the fish exposed to copper is mainly caused by the inhibition on the M-LDH in the fish. 4. The numbers of the esterase isozyme bands of the gill, liver, muscle, blood, brain, and kidney of the normal fish were 3, 6, 2, 2, 2, and 2 respectively, and these numbers were the same as those exposed to copper. The relative mobilities of the esterase bands in the gill, liver, blood, and kidney of the exposed group were different from those of the control. 5. There was one hemoglobin band on the anode in the normal fish. It seems that the nobility of hemoglobin band of the fish exposed to copper was slightly faster than that of the normal fish. 6. The normal gill lamellae of the fish consisted of centrally located pillar cells and a number of mucus cells. When the fish were exposed to copper, the epithelial layer was divorced first, disintegrated, and then destroyed completely. 7. The liver of the normal fish had prominent central veins, cords of hepatic cells, and sinusoids. When the fish were exposed to copper, numerous droplets of fat appeared in the cells around the central vein of the liver. It is assumed that the fatty droplets were accumulated by the lesion due to fatty metamorphosis of the liver caused by copper. 8. There was no histological difference between the muscle of the normal fish and that of the fish exposed to copper. 9. In the normal fish, the tubules of the kidney were surrounded by hemopoetic tissues. However, the kidney tissue of the fish exposed to copper received some damage on the proximal tubules. Since the tubule cells were reduced in height, the lumens of the tubules were enlarged. Consequently many proximal tubules exhibited some pink-stained granular casts and various stages of degeneration.