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Research for biological and biochemical responses during nsh and shellfish cell death processes

nsh 和贝类细胞死亡过程中的生物和生化反应研究

基本信息

批准号：
12460092
负责人：
HIDEAKI Yamanaka
金额：
$ 9.09万
依托单位：
Tokyo University of Fisheries
依托单位国家：
日本
项目类别：
Grant-in-Aid for Scientific Research (B)
财政年份：
2000
资助国家：
日本
起止时间：
2000 至 2002
项目状态：
已结题

来源：
https://kaken.nii.ac.jp/en/grant/KAKENHI-PROJECT-12460092/
关键词：
cell death homeostasis postmortem mitochondria membrane chromatophores norepinephrie hydroperoxide 脂質酸化アポトーシス

项目摘要

It is also true for fish and shellfish that each cell lives after body death. Therefore, metabolism postmortem would be strongly affected by handling and treatment during individual cell living state. The goals of this study is to disclose dynamics of biological and biochemical changes during cell death process and to develop novel methods for evaluating freshness of fish and shellfish. For biological approaches, mitochondrial membrane potentials in fish and crustacean muscle cells excised acutely were determined by the fluorescent probes such as JC-1 and rhodamine123. Then, carp muscle cells remained mitochondrial function during 4-hr anoxia treatment and that re-oxgenation revived mitochondrial membrane potential after 4-hr anoxia treatment, suggesting that carp muscle mitochondria could survive anoxia and synthesize ATP to sustain cell metabolisms. We also observed that crustacean muscle cells maintained the mitochondrial membrane potential 24 hours after body death. Thus, the mitoc … More hondrial membrane potential is a powerful index for evaluating the cell viability and freshness for early stages after whole body death. Red sea bream goes under drastic convulsion 5 - 30 min after cranial spiking. We measured monoamines in the spinal cord during this phenomenon and found that norepinephrine level in the spinal cord was reduced specifically. Then, we concluded that the delayed convulsion of red sea bream would be due to the drastic decrease in spinal cord norepinephrine suppressing spinal cord motoneuron, resulting in the drastic activation of spinal motoneurons. Thus, pinephrine levels in spinal cord and the delayed convulsion are potential indices for freshness for early stages after whole body death. Chromatophore responses were observed under digital microscope immediately after squid body death. Chilling and hypoxia treatment induced the retraction of chromatophore sacs and the responses were maintained over 24 hours after body death. Thus, the chromatophore responses afe also useful for evaluating freshness of squid. We adopted chemiluminescent detection of fatty acid hydroperoxide for the early stage of lipid peroxidation. Dark muscles of yellow tail and horse mackerel exhibited the increase in chemiluminescent due to fatty acid hydroperoxide, which suseested that fatty acid hydroperoxide would be also a potential index for evaluating fish muscle freshness. We also used DPPP, a fluorescent probe for fatty acid hydroperoxide, for evaluating lipid peroxidation of yellow tail and tuna muscle. Consequently, DPPP is also useful for evalulating very early stages of lipid peroxidation of fish muscle.In conclusion, mitochondrial membrane potential, norepinephrine level in spinal cord, delayed convulsion state, chromatophore responses, and fatty acid hydroperoxide levels afe useful and novel indices of freshness for very early stages during postmortem changes. Less

对于鱼类和贝类来说，每个细胞都是在身体死亡后存活的。因此，在单个细胞存活状态下的处理和处置将强烈影响死后代谢。本研究的目的是揭示细胞死亡过程中生物学和生化变化的动力学，并开发新的方法来评价鱼类和贝类的新鲜度。在生物学方法中，用JC-1和罗丹明123等荧光探针测定了急性切除的鱼类和甲壳类动物肌肉细胞的线粒体膜电位。缺氧处理4h后，鲤鱼肌细胞线粒体仍保持正常功能，缺氧处理4h后，复氧恢复线粒体膜电位，说明鲤鱼肌线粒体能在缺氧条件下存活并合成ATP维持细胞代谢。我们还观察到，甲壳动物肌肉细胞在身体死亡后24小时仍保持线粒体膜电位。因此， ...更多信息软骨膜电位是评价全身死亡后早期细胞活力和新鲜度的有力指标。红鲷在头部穿刺后5 - 30分钟发生剧烈抽搐。我们在此现象期间测量了脊髓中的单胺，发现脊髓中的去甲肾上腺素水平特异性降低。因此，我们认为真鲷的迟发性惊厥可能是由于脊髓去甲肾上腺素抑制脊髓运动神经元的急剧减少，导致脊髓运动神经元的急剧激活。因此，脊髓中的松果体素水平和延迟惊厥是全身死亡后早期新鲜的潜在指标。在数码显微镜下观察了鱿鱼尸体死亡后的色素细胞反应。低温和低氧处理引起色素囊的收缩，并且这种反应在尸体死亡后维持超过24小时。因此，色素细胞反应也可用于评价鱿鱼的新鲜度。采用荧光法检测脂质过氧化的早期阶段。黄尾鱼和竹荚鱼的深色肌肉由于脂肪酸过氧化氢的存在，其荧光强度增加，这表明脂肪酸过氧化氢也可能是评价肌肉新鲜度的一个潜在指标。我们还使用了DPPP，一种脂肪酸氢过氧化物的荧光探针，用于评估黄尾金枪鱼和金枪鱼肌肉的脂质过氧化。因此，DPPP也是有用的评价非常早期阶段的脂质过氧化的鱼类肌肉。总之，线粒体膜电位，去甲肾上腺素水平在脊髓，延迟惊厥状态，色素细胞的反应，和脂肪酸氢过氧化物水平是有用的和新的指标新鲜度非常早期阶段在死后的变化。少