title: Characterization of gene expression profiling of mouse tissues obtained during the postmortem interval.
abstract: Attempts to establish a tissue bank from autopsy samples have led to uncovering of the secrets of many diseases. Here, we examined the length of time that the RNA from postmortem tissues is available for microarray analysis and reported the gene expression profile for up- and down-regulated genes during the postmortem interval. We extracted RNA from fresh-frozen (FF) and formalin-fixed paraffin-embedded (FFPE) brains and livers of three different groups of mice: 1) mice immediately after death, 2) mice that were stored at room temperature for 3h after death, and 3) mice that were stored at 4°C for 18h after death, as this storage resembles the human autopsy process in Japan. The RNA quality of the brain and the liver was maintained up to 18h during the postmortem interval. Based on the microarray analysis, we selected genes that were altered by >1.3-fold or <0.77-fold and classified these genes using hierarchical cluster analysis following DAVID gene ontology analysis. These studies revealed that cytoskeleton-related genes were enriched in the set of up-regulated genes, while serine protease inhibitors were enriched in the set of down-regulated genes. Interestingly, although the RNA quality was maintained due to high RNA integrity number (RIN) values, up-regulated genes were not validated by quantitative PCR, suggesting that these genes may become fragmented or modified by an unknown mechanism. Taken together, our findings suggest that under typical autopsy conditions, gene expression profiles that reflect disease pathology can be examined by understanding comprehensive recognition of postmortem fluctuation of gene expression.
authors.affiliation: Department of Biomedical Sciences, College of Life and Health Sciences, Chubu University, Kasugai, Japan.
authors.forename: Sayaka
authors.initials: S
authors.lastname: Sobue
authors.affiliation: Department of Pathology, National Center for Geriatrics and Gerontology, Obu, Japan; Medical Office, Yachimata Reformatory, Yachimata, Japan.
authors.forename: Keita
authors.initials: K
authors.lastname: Sakata
authors.affiliation: Department of Biomedical Sciences, College of Life and Health Sciences, Chubu University, Kasugai, Japan; Department of Pathology, Tokyo Metropolitan Geriatric Hospital, Tokyo, Japan.
authors.forename: Yuki
authors.initials: Y
authors.lastname: Sekijima
authors.affiliation: Department of Biomedical Sciences, College of Life and Health Sciences, Chubu University, Kasugai, Japan.
authors.forename: Shanlou
authors.initials: S
authors.lastname: Qiao
authors.affiliation: Department of Biomedical Sciences, College of Life and Health Sciences, Chubu University, Kasugai, Japan.
authors.forename: Takashi
authors.initials: T
authors.lastname: Murate
authors.affiliation: Department of Biomedical Sciences, College of Life and Health Sciences, Chubu University, Kasugai, Japan. Electronic address: ichihara@isc.chubu.ac.jp.
authors.forename: Masatoshi
authors.initials: M
authors.lastname: Ichihara
mesh_terms: D000818:Animals; D001344:Autopsy / Q000379:methods*; D001921:Brain / Q000378:metabolism; D016000:Cluster Analysis; D003080:Cold Temperature; D005557:Formaldehyde; D020869:Gene Expression Profiling / Q000379:methods*; D063990:Gene Ontology; D006801:Humans; D008099:Liver / Q000378:metabolism; D008297:Male; D008807:Mice, Inbred BALB C; D020411:Oligonucleotide Array Sequence Analysis; D016612:Paraffin Embedding; D011180:Postmortem Changes; D020133:Reverse Transcriptase Polymerase Chain Reaction; D013997:Time Factors; D016707:Tissue Fixation; D014021:Tissue Preservation / Q000379:methods*
chemical_list: D005557:Formaldehyde
keywords: Bioinformatics; Expression profiling; Postmortem interval; RNA degradation