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Effect of ethanol on calcium-uptake and phospholipid turnover in mouse brain and heart synaptosomes
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P.mo.426
ect
ciu
an01 0
Natsuki, R. and Takabatake, E. Faculty of Pharmaceutical Sciences, Setsunan University, 45-l. Nagaotoge-cho. Hirakata. Osaka, 573.01, Japm
The effects of ethanol treatment on mouse brain and heart synaptosomal45calcium (Ca) uptake and phospholipid turnover were investigated. Ethanol was given to mice in drinking water (15%) for 3 weeks. The consumption of ethanol increased gradually during treatment but the food intake was almost as same as control. The body weight of ethanol treated mice was slightly less than that of control. Synaptosomal lipid peroxidation level of ethanol treated tice was higher in the brain than control but lower in heart. On the other hand, synaptosomal no.n-protein thiol level of ethanol treated mice was higher than that of control in either brain or heart. The synaptosomes of either brain or heart were incubated with ethanol, norepinephrine (NE). carbachol (Carb), or isoproterenol (IsoPro) and the uptake of 45Ca and the incorporation of 3-H-inositol or 14-C-choline to phosphatidylinositol (PI) or to posphatidylcholine (PC), respectively, were determined. The 45-Ca uptake of brain and heart from ethanol treated mice were g7 and 216% of control mice, respectively. Not only ethanol but aiso NE, Garb, or IsoPro added in vitro increased 45-Ca uptake in all the cases. However, the low dose of ethanol (100 mM) increased 45-Ca uptake by brain of either control or ethanol treated mice, but the high dose (500 mM) decreased them. The incorporation of 3-H-inositol into PI in the brain and heart synaptosomes of ethanol treated mice were 150 and 113% of that of control, respectively. On the other hand, the incorporation of 14C-choline into PC in the brain and heart of ethanol treated mice were 104 and 126% of control, respectively. The in vitro addition of ethanol, NE, Garb, or IsoPro in brain synaptosomes increased the incorporation of 3-H-inositol and 14Gcholine into PI and PC, respectively, in either control or ethanol treated mice. In the case of heart synaptosomes, NE and Carb increased the incorporation of 3-H-inositol and 14-C-choline into phosphoiipids in control mice but not in ethanol treated mice. However, IsoPro increased significantly these incorporation by either control or ethanol treated mice heart synaptosomes. These results suggest that alpha-adrenoceptors and choline& system of heart could play any important role in modulating the toxic effects of ethanol. Table 1 The incorporation of 14C-choline into brain and heart synaptosomal. Tissue
Treatment
None
Chemicals added in vitro
Ethanol Brain Ethanol Heart Ethanol
Control 104i 3 Control 126f 7
100+ 4 121& 5 * 100*14 108&15
113f 4* 120+ 13 * lll& 7 117+19
NE
Carb
IsoPro
118* 6 * 128f 2* 168k13 * 127+- 5
117f 8 * 154&12 * 177& 13 * 174+11 *
241+14 * 179f
4 *
The synaptosomes of brain or heart were incubated with ethanol (100 mM), NE (100 pM), Carb (100 PM), or IsoPro (100 PM) and WC-choline (20 PM) for 20 min at 37°C. Phospholipids were extracted with chloroform-methanol (2 : 1) and the radioactivities were measured. Data from 5 experiments are expressed as S of the none addition group of control mice in each tissue. * Significantly different from the none addition group of each treatment at p < 0.01.
I
P.mo.426
ect
ciu
an01 0
Natsuki, R. and Takabatake, E. Faculty of Pharmaceutical Sciences, Setsunan University, 45-l. Nagaotoge-cho. Hirakata. Osaka, 573.01, Japm
The effects of ethanol treatment on mouse brain and heart synaptosomal45calcium (Ca) uptake and phospholipid turnover were investigated. Ethanol was given to mice in drinking water (15%) for 3 weeks. The consumption of ethanol increased gradually during treatment but the food intake was almost as same as control. The body weight of ethanol treated mice was slightly less than that of control. Synaptosomal lipid peroxidation level of ethanol treated tice was higher in the brain than control but lower in heart. On the other hand, synaptosomal no.n-protein thiol level of ethanol treated mice was higher than that of control in either brain or heart. The synaptosomes of either brain or heart were incubated with ethanol, norepinephrine (NE). carbachol (Carb), or isoproterenol (IsoPro) and the uptake of 45Ca and the incorporation of 3-H-inositol or 14-C-choline to phosphatidylinositol (PI) or to posphatidylcholine (PC), respectively, were determined. The 45-Ca uptake of brain and heart from ethanol treated mice were g7 and 216% of control mice, respectively. Not only ethanol but aiso NE, Garb, or IsoPro added in vitro increased 45-Ca uptake in all the cases. However, the low dose of ethanol (100 mM) increased 45-Ca uptake by brain of either control or ethanol treated mice, but the high dose (500 mM) decreased them. The incorporation of 3-H-inositol into PI in the brain and heart synaptosomes of ethanol treated mice were 150 and 113% of that of control, respectively. On the other hand, the incorporation of 14C-choline into PC in the brain and heart of ethanol treated mice were 104 and 126% of control, respectively. The in vitro addition of ethanol, NE, Garb, or IsoPro in brain synaptosomes increased the incorporation of 3-H-inositol and 14Gcholine into PI and PC, respectively, in either control or ethanol treated mice. In the case of heart synaptosomes, NE and Carb increased the incorporation of 3-H-inositol and 14-C-choline into phosphoiipids in control mice but not in ethanol treated mice. However, IsoPro increased significantly these incorporation by either control or ethanol treated mice heart synaptosomes. These results suggest that alpha-adrenoceptors and choline& system of heart could play any important role in modulating the toxic effects of ethanol. Table 1 The incorporation of 14C-choline into brain and heart synaptosomal. Tissue
Treatment
None
Chemicals added in vitro
Ethanol Brain Ethanol Heart Ethanol
Control 104i 3 Control 126f 7
100+ 4 121& 5 * 100*14 108&15
113f 4* 120+ 13 * lll& 7 117+19
NE
Carb
IsoPro
118* 6 * 128f 2* 168k13 * 127+- 5
117f 8 * 154&12 * 177& 13 * 174+11 *
241+14 * 179f
4 *
The synaptosomes of brain or heart were incubated with ethanol (100 mM), NE (100 pM), Carb (100 PM), or IsoPro (100 PM) and WC-choline (20 PM) for 20 min at 37°C. Phospholipids were extracted with chloroform-methanol (2 : 1) and the radioactivities were measured. Data from 5 experiments are expressed as S of the none addition group of control mice in each tissue. * Significantly different from the none addition group of each treatment at p < 0.01.