| Project/Area Number |
12460092
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| Research Category |
Grant-in-Aid for Scientific Research (B)
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| Allocation Type | Single-year Grants |
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| Section | 一般 |
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| Research Field |
Fisheries chemistry
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| Research Institution | Tokyo University of Fisheries |
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Principal Investigator |
HIDEAKI Yamanaka Tokyo Univ. Fish., Fd. Sci. & Technol., Professor, 水産学部, 教授 (20092596)
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| Co-Investigator(Kenkyū-buntansha) |
潮 秀樹 東京水産大学, 水産学部, 助教授 (50251682)
齋藤 洋昭 (独)水産総合研究センター, 中央水産研究所, 室長(研究職)
TOSHIAKI Ohshima Tokyo Univ. Fish., Fd. Sci. & Technol.. Assoc. Professor, 水産学部, 助教授 (70134856)
HIROAKI Saito Natl. Res. Inst. Fish. Sci., Fish. Res. Agency
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| Project Period (FY) |
2000 – 2002
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| Project Status |
Completed (Fiscal Year 2002)
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| Budget Amount *help |
¥14,200,000 (Direct Cost: ¥14,200,000)
Fiscal Year 2002: ¥3,400,000 (Direct Cost: ¥3,400,000)
Fiscal Year 2001: ¥3,700,000 (Direct Cost: ¥3,700,000)
Fiscal Year 2000: ¥7,100,000 (Direct Cost: ¥7,100,000)
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| Keywords | cell death / homeostasis / postmortem / mitochondria / membrane / chromatophores / norepinephrie / hydroperoxide / 脂質酸化 / アポトーシス |
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| Research Abstract |
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
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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
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