Improved RNA analysis for immediate autopsy of temporal bone soft tissues

RNA analysis is essential for understanding biological activities of a cell or tissue. Unfortunately, retrieval of RNA from existing archives of human temporal bones has proven extremely difficult due to degradation of RNA mo lecules. The major factors that contribute to degradation of RNA in specime ns from autopsied temporal bones are tissue autolysis due to time elapsed b efore autopsy. and technical problems in processing the bones after harvest . We therefore focused oil improving the survival of RNA in human temporal bones by shortening the time to autopsy and through modification of the pro cessing technique by removing targeted tissues directly from the temporal b ones and by avoiding time-consuming decalcification and celloidin-embedding . Eight temporal bones collected at immediate autopsies were used in this s tudy. Representative mRNAs, ranging From high (MUC5B, physically unstable) to low (beta-actin, physically stable) molecular weights. and from abundant (MUC5B) to non-abundant (MUC1) RNA, were studied by in situ hybridization, Northern blot technique, or both. Using this modified protocol in autopsie s performed up to 6 h after death. the existence of mRNAs was demonstrated in all bones studied. This improved method demonstrates the feasibility of the List of autopsied temporal bone tissues for RNA analysis.