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Electro microscopic observation of mice hearts administered methamphetamine
j Mol Cell Cardiol 19 (Supplement 1) (1987)
91 ULTRASTRUCTURAL STUDY OF THE MYOCARDIUM 1N EXPERIMENTAL DIABETIC MYOCARDIOPATHY; EFFECT OF PANCREATIC TRANSPLANTATION. T. Hayashi, H. Deguehi, Y. Kitaura, K. Kawamura, *M. Nozawa. The 3rd Div. Dept. Int. Med., Osaka Medical College, Takatsuki, *Dept. Surgery, Josai Dental University, Saitama, Japan. We studied ultrastruetural changes of the myocardium in diabetic rats and tried to evaluate the effect of pancreatic transplantation on diabetic myocardiopathy. Male Lewis rats were divided into three groups (4 rats each): 1) normal, 2) diabetic rats induced by intravenous administration of streptozotocin (STZ) 60 mg/Kg, and 3) STZ induced diabetic rats with pancreatic transplantation. All animals were fed for 2 months, and were then examined. Twelve to fifteen blood capillaries from each tissue block were photographed in the electron microscope, and thickness of the basement membrane of the vessels was estimated by Williamson's method. In group 2, the blood sugar elevated over 300 mg/dl, and the body and heart weights significantly decreased; but they were all improved in group 3. Electron Microscopy: In group 1, the myoeytes and capillaries appeared to be normal in the myocardium. In group 2, various morphological changes were seen, including a thickening in the basal lamina of capillaries, an accumulation of lipid droplets, vacuolar formation and an increased number of myelin figures in many eardiocytes. In group 3, the above changes were very much smaller in size and degree. These findings show that pancreatic transplantation decreases the incidence and severity of cardiac ultrastructural changes in diabetic rats.
92
C I R C U L A T I N G A U T 0 - A N T I B O D I E S A G A I N S T T H E M E M B R A N E OF I S O L A T E D C A R D I O C Y T E L I G H T A N D E L E C T R O N M I C R O S C O P I C A L STUDY. T. I z u m i , B. M a i s c h a n d K. K o c h s i e k . D e p a r t m e n t of Internal Medicine, Niigata University School of Medicine, Niigata 951, Japan and U n i v e r s i t y of W u e r z b u r g , W u e r z b u r g 8700, FR. G e r m a n y . C i r c u l a t i n g a u t o - a n t i b o d i e s a g a i n s t the m e m b r a n e o f i s o l a t e d c a r d i o cyte: antimyolemmal antibodies(AMLAs) are likely the most relevant humoral diagnostic m a r k e r in t h e i n f l a m m a t o r y heart disease. In this study AMLAs are analized by light and electron microscopy for their b i n d i n g site. ( M a t e r i a l s a n d m e t h o d s ) F r o m m o r e t h a n 5 0 0 0 p a t i e n t s 16 selected sera with more than 1:80 in AMLA titers were investigated. Isolated human, rat and mouse cardiocytes w e r e u s e d as t a r g e t c e l l s . F l u o r e s c e n c e m i c r o s c o p y w a s p e r f o r m e d a c c o r d i n g to the i n d i r e c t m e t h o d . Immunoelectron microscopically the rat cardiocyte was treated with avidin-biotin complex after 4% buffered paraform aldehyde(pH 7.4). ( R e s u l t s ) I. T h e h u m a n a n d r a t c a r d i o c y t e revealed positive linear staining on t h e c e l l s u r f a c e , b u t i n m o u s e t h e f o r m is r o u n d . 2. T h e reduction in A M L A s t i t e r b y a b s o r p t i o n was achieved at 100,000 per m i c r o l i t e r in c a r d i o c y t e c o n c e n t r a t i o n at least. 3. E l e c t r o n m i c r o s c o p i c a l l y , D A B p a r t i c l e b i n d s to the s u b s a r c o l e m m a l area. -
93
ELECTRO MICROSCOPIC OBSERVATION OF MICE HEARTS ADMINISTERED METHAMPHETAMINE Ryoji Matoba, Shunzo Onishi ~ Noboru Fujitani, Ichiro Shikata. Department of Legal Medicine, Osaka University Medical School. ~ of Biomedical Technology, Osaka Univ. We have already reported that there are various cardiac legions among methamphetamine (MP) ab~isers and that these changes are also seen on the mouse hearts being administered MP for a relatively long time. To investigate these mechanism of cardiac lesions, we researched these changes mainly by using EM. Male DDY mice (4 weeks old) were injected MP 20 mg/g b.w. subcutaneously every day for 2 weeks and 4 weeks. As a control, same volume saline wer e injected. Every groups were consisted of 5 mice. Two days after last injection, animals were anesthetized with pentobarbital and were removed the hearts. They were cut into 1 mm cube blocks of the left ventricle, were fixed in the 2% glutar -a]dehvde in 0.I M phosphate buffer, post-fixed with 1% osminium tetroxide, dehydrated in graded alcohols, and embedded in Epon 812. In the MP administered mice, these findings were frequently seen : hypertrophy,super-contraction, disarray and unclarification of myofibrils. Degeneration, swellings, cramps of mitochondria and dilatation of sarcoplasmic reticulum especially just under the sarcolemma. Sometimes these vesicles which were thought to be sarcoplasmic reticulums were seen to be excluded to the extra-cellular space as " exocytosis." This findings are interesting thinking that catecholamine release by MP will result in calcium overload on cardiac cells. There were not so much differences between 2w and 4w, but these were scarcely seen in the control groups.
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91 ULTRASTRUCTURAL STUDY OF THE MYOCARDIUM 1N EXPERIMENTAL DIABETIC MYOCARDIOPATHY; EFFECT OF PANCREATIC TRANSPLANTATION. T. Hayashi, H. Deguehi, Y. Kitaura, K. Kawamura, *M. Nozawa. The 3rd Div. Dept. Int. Med., Osaka Medical College, Takatsuki, *Dept. Surgery, Josai Dental University, Saitama, Japan. We studied ultrastruetural changes of the myocardium in diabetic rats and tried to evaluate the effect of pancreatic transplantation on diabetic myocardiopathy. Male Lewis rats were divided into three groups (4 rats each): 1) normal, 2) diabetic rats induced by intravenous administration of streptozotocin (STZ) 60 mg/Kg, and 3) STZ induced diabetic rats with pancreatic transplantation. All animals were fed for 2 months, and were then examined. Twelve to fifteen blood capillaries from each tissue block were photographed in the electron microscope, and thickness of the basement membrane of the vessels was estimated by Williamson's method. In group 2, the blood sugar elevated over 300 mg/dl, and the body and heart weights significantly decreased; but they were all improved in group 3. Electron Microscopy: In group 1, the myoeytes and capillaries appeared to be normal in the myocardium. In group 2, various morphological changes were seen, including a thickening in the basal lamina of capillaries, an accumulation of lipid droplets, vacuolar formation and an increased number of myelin figures in many eardiocytes. In group 3, the above changes were very much smaller in size and degree. These findings show that pancreatic transplantation decreases the incidence and severity of cardiac ultrastructural changes in diabetic rats.
92
C I R C U L A T I N G A U T 0 - A N T I B O D I E S A G A I N S T T H E M E M B R A N E OF I S O L A T E D C A R D I O C Y T E L I G H T A N D E L E C T R O N M I C R O S C O P I C A L STUDY. T. I z u m i , B. M a i s c h a n d K. K o c h s i e k . D e p a r t m e n t of Internal Medicine, Niigata University School of Medicine, Niigata 951, Japan and U n i v e r s i t y of W u e r z b u r g , W u e r z b u r g 8700, FR. G e r m a n y . C i r c u l a t i n g a u t o - a n t i b o d i e s a g a i n s t the m e m b r a n e o f i s o l a t e d c a r d i o cyte: antimyolemmal antibodies(AMLAs) are likely the most relevant humoral diagnostic m a r k e r in t h e i n f l a m m a t o r y heart disease. In this study AMLAs are analized by light and electron microscopy for their b i n d i n g site. ( M a t e r i a l s a n d m e t h o d s ) F r o m m o r e t h a n 5 0 0 0 p a t i e n t s 16 selected sera with more than 1:80 in AMLA titers were investigated. Isolated human, rat and mouse cardiocytes w e r e u s e d as t a r g e t c e l l s . F l u o r e s c e n c e m i c r o s c o p y w a s p e r f o r m e d a c c o r d i n g to the i n d i r e c t m e t h o d . Immunoelectron microscopically the rat cardiocyte was treated with avidin-biotin complex after 4% buffered paraform aldehyde(pH 7.4). ( R e s u l t s ) I. T h e h u m a n a n d r a t c a r d i o c y t e revealed positive linear staining on t h e c e l l s u r f a c e , b u t i n m o u s e t h e f o r m is r o u n d . 2. T h e reduction in A M L A s t i t e r b y a b s o r p t i o n was achieved at 100,000 per m i c r o l i t e r in c a r d i o c y t e c o n c e n t r a t i o n at least. 3. E l e c t r o n m i c r o s c o p i c a l l y , D A B p a r t i c l e b i n d s to the s u b s a r c o l e m m a l area. -
93
ELECTRO MICROSCOPIC OBSERVATION OF MICE HEARTS ADMINISTERED METHAMPHETAMINE Ryoji Matoba, Shunzo Onishi ~ Noboru Fujitani, Ichiro Shikata. Department of Legal Medicine, Osaka University Medical School. ~ of Biomedical Technology, Osaka Univ. We have already reported that there are various cardiac legions among methamphetamine (MP) ab~isers and that these changes are also seen on the mouse hearts being administered MP for a relatively long time. To investigate these mechanism of cardiac lesions, we researched these changes mainly by using EM. Male DDY mice (4 weeks old) were injected MP 20 mg/g b.w. subcutaneously every day for 2 weeks and 4 weeks. As a control, same volume saline wer e injected. Every groups were consisted of 5 mice. Two days after last injection, animals were anesthetized with pentobarbital and were removed the hearts. They were cut into 1 mm cube blocks of the left ventricle, were fixed in the 2% glutar -a]dehvde in 0.I M phosphate buffer, post-fixed with 1% osminium tetroxide, dehydrated in graded alcohols, and embedded in Epon 812. In the MP administered mice, these findings were frequently seen : hypertrophy,super-contraction, disarray and unclarification of myofibrils. Degeneration, swellings, cramps of mitochondria and dilatation of sarcoplasmic reticulum especially just under the sarcolemma. Sometimes these vesicles which were thought to be sarcoplasmic reticulums were seen to be excluded to the extra-cellular space as " exocytosis." This findings are interesting thinking that catecholamine release by MP will result in calcium overload on cardiac cells. There were not so much differences between 2w and 4w, but these were scarcely seen in the control groups.
S.53