Research on hypoxic sudden death by analysis of mitochondrial respiratory-chain-related DNA

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In forensic autopsy, death from suffocation is extremely difficult to diagnose. So, technique of molecular biology is indispensable for elucidation of the cause of death.Mitochondria has DNAs as well as intra-nuclear DNA, in addition, it is expected that the variation is caused in mitochondrial DNA under a hypoxic condition. Therefore, I tried to do general analysis of a respiratory-chain-affiliated-domain of mitochondria DNA, in myocardium and intra-cerebral cells, by the detection of a manifestation of a respiratory chain-affiliated-DNA of mitochondria, in the relation with polymorphism with hypoxic sudden death cases. I collected tissue samples of the myocardium with a frozen specimen for the identification of death from suffocation or a respiratory organs system disorder or acute heart death and hypoxia by a circulatory organ system disorder. PCR were performed using a single cell by micro-dissection-system.In addition, I did a direct sequence of a respiratory-chain-related domain and analyzed a sequence of mitochondria DNA to detect SNPs (single base variation).The good results was obtained in the cases relatively long time was passed from asphyxiation to death, in addition, remarkable results were provided with a heart and a kidney. However, as for a brain, usefulness as a sample is expected because high sensitivity of hypoxia, but collapse of a cell was early, and deviation of data was found.In real time PCR, it became clear that two step of PCR method was indispensable for detections.I made contribution to apply DNA technology to forensic medicine in cases of sudden death and a diagnosis of death from suffocation by a change of mitochondria DNA. By this study, it is possible to push forward discussion about the origin of mitochondrial DNA.