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

对于鱼类和贝类来说，每个细胞在身体死亡后都会存活，这一点也是正确的。因此，在个体细胞生存状态下，处理和处理对死后新陈代谢有很大的影响。本研究的目的是揭示细胞死亡过程中生物和生化变化的动态，并开发新的方法来评价鱼类和贝类的新鲜度。对于生物学方法，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肌肉细胞的线粒体膜电位。然后，鲤鱼肌肉细胞在4小时缺氧处理中仍保持线粒体功能，缺氧4小时后复氧使线粒体膜电位恢复，这表明鲤鱼肌肉线粒体可以在缺氧中存活并合成ATP以维持细胞代谢。我们还观察到甲壳动物肌肉细胞在身体死亡24小时后维持线粒体膜电位。因此，Mitoc…更多的软骨细胞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。罗非鱼在颅尖后5-30分钟发生剧烈惊厥。在这一现象期间，我们测量了脊髓中的单胺类物质，发现脊髓中去甲肾上腺素的水平特别降低。由此得出结论，罗非鱼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，导致脊髓运动神经元的剧烈激活所致。因此，脊髓中的肾上腺素水平和迟发性惊厥是全身死亡后早期新鲜度的潜在指标。在鱿鱼身体死亡后即刻在数码显微镜下观察发色团反应。低温和低氧处理引起发色团囊壁收缩，这种反应在体死后24小时内保持不变。因此，发色团响应AFE也可用于评价鱿鱼的新鲜度。在脂质过氧化的早期阶段，我们采用化学发光法检测脂肪酸氢过氧化。脂肪酸氢过氧化氢增加了黄尾鱼和马鱼肌肉的化学发光强度，表明脂肪酸氢过氧化氢也可能是评价鱼肉新鲜度的一个潜在指标。我们还使用了脂肪酸氢过氧化氢的荧光探针DPPP来评估黄尾和金枪鱼肌肉的脂质过氧化作用。总而言之，线粒体膜电位、脊髓去甲肾上腺素水平、迟发性惊厥状态、生色团反应和脂肪酸氢过氧化氢水平在死后变化的非常早期阶段是有用的和新的新鲜度指标。较少

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