Estimating time of death based on the biological clock

The biological clock may stop at the time of death in a dead body. Therefore, the biological clock seems useful for estimating the time of death. In this study, we tried to read the biological clock in tissues from dead bodies to estimate the time of death using molecular biological techniques. At first, we examined real-time RT-PCR analysis of gene expression for mPer2 and mBmal1, which constitutes a feedback loop in the oscillation system, in the kidney, liver, and heart of mice. We could detect circadian oscillation of these gene expressions in mouse tissues even at < 48 h after death. Thus, the ratio of mPer2/mBmal1 was found to be useful for estimating the time of death. We next applied this method to the liver, kidney, and heart obtained from forensic autopsy cases with less than 72 h of postmortem interval. Significant circadian oscillation of hPer2/hBmal1 ratio could be detected in these autopsy samples. We further examined gene expression for hRev-Erb alpha, a component of another feedback loop. The ratios of hRev-Erb alpha/hBmal1 showed higher amplitude of oscillation than those of hPer2/hBmal1 and are considered more suitable for estimating the time of death. In particular, a hRev/hBmal1 ratio of > 50 indicated the time of death as 0200-0900 hours, and a hRev/hBmal1 ratio that considerably exceeded 75 indicated the time of death as 0200-0800 hours. On the other hand, a hRev/hBmal1 ratio of less than 25 strongly indicated the time of death as 1000-2300 hours. Taken together, these findings indicate that gene expression analyses of the biological clock could be powerful methods for estimation of the time of death.