{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,10,9]],"date-time":"2026-10-09T19:54:45Z","timestamp":1791575685922,"version":"4.3.4"},"reference-count":29,"publisher":"MDPI AG","issue":"15","license":[{"start":{"date-parts":[[2025,7,22]],"date-time":"2025-07-22T00:00:00Z","timestamp":1753142400000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"name":"University Innovation Support Project through the National Research Foundation of Korea funded by the Ministry of Education"}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["IJMS"],"abstract":"<jats:p>The postmortem interval (PMI), defined as the time elapsed between death and the discovery or examination of the body, is a crucial parameter in forensic science for estimating the time of death. There are many ways to measure the PMI, such as Henssge\u2019s nomogram, which uses rectal temperature measurement; livor mortis; rigor mortis; and forensic entomology. However, these methods are usually affected by various conditions in the surrounding environment. The purpose of the present study was to compare molecular genetics and histological changes in the brain and skeletal muscle tissues of SD rats over increasing periods of time after death. For the PMIs, we considered 0 h, 6 h, 12 h, 24 h, 36 h, 48 h, 4 days, 6 days, 8 days, 10 days, 14 days, and 21 days and compared them at 4 \u00b0C and 26 \u00b0C. Hematoxylin and Eosin (H&amp;E) staining was performed to observe tissue changes. Morphological tissue changes were observed in cells for up to 21 days at 4 \u00b0C, and cell destruction was visually confirmed after 14 days at 26 \u00b0C. Total RNA (tRNA) was isolated from each tissue sample, and complementary DNA (cDNA) was synthesized. A reverse transcription quantitative PCR (RT-qPCR) SYBR Green assay targeting three types of housekeeping genes, including Gapdh, Sort1, B2m, and 5S rRNA, was performed. The results showed that Gapdh and 5S rRNA were highly stable and could be better RNA targets for estimating the PMI in brain and skeletal muscle tissues. Conversely, Sort1 and B2m showed poor stability and low expression levels. In conclusion, these molecular biomarkers could be used as auxiliary indicators of the PMI in human, depending on the stability of the marker.<\/jats:p>","DOI":"10.3390\/ijms26157059","type":"journal-article","created":{"date-parts":[[2025,7,22]],"date-time":"2025-07-22T15:00:42Z","timestamp":1753196442000},"page":"7059","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":6,"title":["Postmortem Changes in mRNA Expression and Tissue Morphology in Brain and Femoral Muscle Tissues of Rat"],"prefix":"10.3390","volume":"26","author":[{"ORCID":"https:\/\/orcid.org\/0009-0000-0135-4693","authenticated-orcid":false,"given":"Sujin","family":"Choi","sequence":"first","affiliation":[{"name":"Department of Forensic Science, Graduate School, Catholic University of Pusan, Busan 46252, Republic of Korea"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-6916-4498","authenticated-orcid":false,"given":"Minju","family":"Jung","sequence":"additional","affiliation":[{"name":"Department of Clinical Laboratory Science, College of Health Sciences, Catholic University of Pusan, Busan 46252, Republic of Korea"},{"name":"Next-Generation Industrial Field-Based Specialist Program for Molecular Diagnostics, Brain Busan 21 Plus Program, Graduate School, Catholic University of Pusan, Busan 46252, Republic of Korea"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Mingyoung","family":"Jeong","sequence":"additional","affiliation":[{"name":"Department of Forensic Science, Graduate School, Catholic University of Pusan, Busan 46252, Republic of Korea"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-2585-1226","authenticated-orcid":false,"given":"Sohyeong","family":"Kim","sequence":"additional","affiliation":[{"name":"Department of Clinical Laboratory Science, College of Health Sciences, Catholic University of Pusan, Busan 46252, Republic of Korea"},{"name":"Next-Generation Industrial Field-Based Specialist Program for Molecular Diagnostics, Brain Busan 21 Plus Program, Graduate School, Catholic University of Pusan, Busan 46252, Republic of Korea"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Dong Geon","family":"Lee","sequence":"additional","affiliation":[{"name":"Department of Clinical Laboratory Science, College of Health Sciences, Catholic University of Pusan, Busan 46252, Republic of Korea"},{"name":"Next-Generation Industrial Field-Based Specialist Program for Molecular Diagnostics, Brain Busan 21 Plus Program, Graduate School, Catholic University of Pusan, Busan 46252, Republic of Korea"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-4022-3415","authenticated-orcid":false,"given":"Kwangmin","family":"Park","sequence":"additional","affiliation":[{"name":"Department of Clinical Laboratory Science, College of Health Sciences, Catholic University of Pusan, Busan 46252, Republic of Korea"},{"name":"Next-Generation Industrial Field-Based Specialist Program for Molecular Diagnostics, Brain Busan 21 Plus Program, Graduate School, Catholic University of Pusan, Busan 46252, Republic of Korea"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Xianglan","family":"Xuan","sequence":"additional","affiliation":[{"name":"Department of Clinical Laboratory Science, College of Health Sciences, Catholic University of Pusan, Busan 46252, Republic of Korea"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Heechul","family":"Park","sequence":"additional","affiliation":[{"name":"Department of Clinical Laboratory Science, Hyejeon University, Hongseong 32244, Republic of Korea"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-5147-1783","authenticated-orcid":false,"given":"Dong Hyeok","family":"Kim","sequence":"additional","affiliation":[{"name":"Department of Biomedical Laboratory Science, Masan University, Changwon 51217, Republic of Korea"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Jungho","family":"Kim","sequence":"additional","affiliation":[{"name":"Department of Forensic Science, Graduate School, Catholic University of Pusan, Busan 46252, Republic of Korea"},{"name":"Department of Clinical Laboratory Science, College of Health Sciences, Catholic University of Pusan, Busan 46252, Republic of Korea"},{"name":"Next-Generation Industrial Field-Based Specialist Program for Molecular Diagnostics, Brain Busan 21 Plus Program, Graduate School, Catholic University of Pusan, Busan 46252, Republic of Korea"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Min Ho","family":"Lee","sequence":"additional","affiliation":[{"name":"Forensic DNA Division, Gwangju Institute, National Forensic Service, Jangseong 57248, Republic of Korea"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Yoonjung","family":"Cho","sequence":"additional","affiliation":[{"name":"Forensic DNA Division, Gwangju Institute, National Forensic Service, Jangseong 57248, Republic of Korea"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-2511-6555","authenticated-orcid":false,"given":"Sunghyun","family":"Kim","sequence":"additional","affiliation":[{"name":"Department of Forensic Science, Graduate School, Catholic University of Pusan, Busan 46252, Republic of Korea"},{"name":"Department of Clinical Laboratory Science, College of Health Sciences, Catholic University of Pusan, Busan 46252, Republic of Korea"},{"name":"Next-Generation Industrial Field-Based Specialist Program for Molecular Diagnostics, Brain Busan 21 Plus Program, Graduate School, Catholic University of Pusan, Busan 46252, Republic of Korea"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"1968","published-online":{"date-parts":[[2025,7,22]]},"reference":[{"key":"ref_1","unstructured":"Kang, D.Y., Kang, H.U., Gwak, J.S., Kim, J.H., Park, U.I.U., Park, J.T., Park, H.G., Lee, S.Y., Lee, S.H., and Lee, S.D. (2007). A Textbook of Legal Medicine, Jungmunkag. [1st ed.]."},{"key":"ref_2","doi-asserted-by":"crossref","unstructured":"Sampaio-Silva, F., Magalh\u00e3es, T., Carvalho, F., Dinis-Oliveira, R.J., and Silvestre, R. (2013). Profiling of RNA degradation for estimation of post mortem [corrected] interval. PLoS ONE, 8.","DOI":"10.1371\/annotation\/cb8b37ac-cbe2-45e3-b9fe-d62e7ced4b25"},{"key":"ref_3","doi-asserted-by":"crossref","unstructured":"Cianci, V., Mondello, C., Sapienza, D., Guerrera, M.C., Cianci, A., Crac\u00f2, A., Luppino, F., Gioffr\u00e8, V., Gualniera, P., and Asmundo, A. (2024). microRNAs as New Biomolecular Markers to Estimate Time since Death: A Systematic Review. Int. J. Mol. Sci., 25.","DOI":"10.3390\/ijms25179207"},{"key":"ref_4","doi-asserted-by":"crossref","first-page":"451","DOI":"10.1007\/s12024-016-9776-y","article-title":"Methods for determining time of death","volume":"12","author":"Madea","year":"2016","journal-title":"Forensic Sci. Med. Pathol."},{"key":"ref_5","doi-asserted-by":"crossref","first-page":"102049","DOI":"10.1016\/j.jflm.2020.102049","article-title":"Estimation of the post-mortem interval using microRNA in the bones","volume":"75","author":"Na","year":"2020","journal-title":"J. Forensic Leg. Med."},{"key":"ref_6","doi-asserted-by":"crossref","unstructured":"Dachet, F., Brown, J.B., Valyi-Nagy, T., Narayan, K.D., Serafini, A., Boley, N., Gingeras, T.R., Celniker, S.E., Mohapatra, G., and Loeb, J.A. (2021). Selective time-dependent changes in activity and cell-specific gene expression in human postmortem brain. Sci. Rep., 11.","DOI":"10.1038\/s41598-021-85801-6"},{"key":"ref_7","doi-asserted-by":"crossref","unstructured":"Pozhitkov, A.E., Neme, R., Domazet-Lo\u0161o, T., Leroux, B.G., Soni, S., Tautz, D., and Noble, P.A. (2017). Tracing the dynamics of gene transcripts after organismal death. Open Biol., 7.","DOI":"10.1098\/rsob.160267"},{"key":"ref_8","first-page":"16","article-title":"Histological changes in lingual striated muscle tissue of human cadavers to estimate the postmortem interval","volume":"19","author":"Fors","year":"2023","journal-title":"Forensic Sci. Med. Pathol."},{"key":"ref_9","doi-asserted-by":"crossref","first-page":"311","DOI":"10.1007\/s10561-010-9210-8","article-title":"Postmortem interval effect on RNA and gene expression in human brain tissue","volume":"12","author":"Birdsill","year":"2011","journal-title":"Cell Tissue Bank."},{"key":"ref_10","doi-asserted-by":"crossref","unstructured":"Joshi, C.J., Ke, W., Drangowska-Way, A., O\u2019Rourke, E.J., and Lewis, N.E. (2022). What are housekeeping genes?. PLoS Comput. Biol., 18.","DOI":"10.1371\/journal.pcbi.1010295"},{"key":"ref_11","doi-asserted-by":"crossref","first-page":"332","DOI":"10.2144\/00292rv02","article-title":"Control selection for RNA quantitation","volume":"29","author":"Suzuki","year":"2000","journal-title":"Biotechniques"},{"key":"ref_12","doi-asserted-by":"crossref","first-page":"498","DOI":"10.1111\/j.1365-2990.2010.01098.x","article-title":"Human brain weight is correlated with expression of the housekeeping genes\u2019 beta-2-microglobulin (\u03b22M) and TATA-binding protein (TBP)","volume":"36","author":"Harrison","year":"2010","journal-title":"Neuropathol. Appl. Neurobiol."},{"key":"ref_13","doi-asserted-by":"crossref","first-page":"92","DOI":"10.1002\/jnr.20231","article-title":"Neurotensin receptor-3\/sortilin mediates neurotensin- induced cytokine\/chemokine expression in a murine microglial cell line","volume":"78","author":"Dicou","year":"2004","journal-title":"J. Neurosci. Res."},{"key":"ref_14","doi-asserted-by":"crossref","first-page":"191","DOI":"10.18388\/abp.2001_5126","article-title":"Structure and functions of 5S rRNA","volume":"48","author":"Barciszewska","year":"2001","journal-title":"Acta. Biochim. Pol."},{"key":"ref_15","doi-asserted-by":"crossref","unstructured":"Taki, F.A., Abdel-Rahman, A.A., and Zhang, B. (2014). A Comprehensive Approach to Identify Reliable Reference Gene Candidates to Investigate the Link between Alcoholism and Endocrinology in Sprague-Dawley Rats. PLoS ONE, 9.","DOI":"10.1371\/journal.pone.0094311"},{"key":"ref_16","doi-asserted-by":"crossref","first-page":"302","DOI":"10.1002\/syn.20496","article-title":"Reference genes for normalization: A study of rat brain tissue","volume":"62","author":"Bonefeld","year":"2008","journal-title":"Synapse"},{"key":"ref_17","first-page":"363","article-title":"RNA Analysis in Forensic Molecular Biology","volume":"121","author":"Courts","year":"2024","journal-title":"Dtsch. Arztebl. Int."},{"key":"ref_18","doi-asserted-by":"crossref","first-page":"307","DOI":"10.1016\/j.scijus.2014.03.001","article-title":"Postmortem interval determination using 18S-rRNA and microRNA","volume":"54","author":"Li","year":"2014","journal-title":"Sci. Justice"},{"key":"ref_19","doi-asserted-by":"crossref","first-page":"628","DOI":"10.2144\/04364ST03","article-title":"Evaluating RNA status for RT -PCR in extracts of postmortem human brain tissue","volume":"36","author":"Miller","year":"2004","journal-title":"Biotechniques"},{"key":"ref_20","doi-asserted-by":"crossref","first-page":"e14221","DOI":"10.7717\/peerj.14221","article-title":"Comparison of reference gene expression stability in mouse skeletal muscle via five algorithms","volume":"10","author":"Ma","year":"2022","journal-title":"PeerJ"},{"key":"ref_21","doi-asserted-by":"crossref","first-page":"149","DOI":"10.1007\/s00414-019-02199-7","article-title":"Postmortem interval determination using mRNA markers ana DNA normalization","volume":"134","author":"Peng","year":"2020","journal-title":"Int. J. Leg. Med."},{"key":"ref_22","doi-asserted-by":"crossref","first-page":"1","DOI":"10.1111\/j.1471-4159.1993.tb03532.x","article-title":"Pre- and postmortem influences on brain RNA","volume":"61","author":"Barton","year":"1993","journal-title":"J. Neurochem."},{"key":"ref_23","doi-asserted-by":"crossref","first-page":"1629","DOI":"10.1007\/s00414-019-02125-x","article-title":"Analysis of RNA in the estimation of post-mortem interval: A review of current evidence","volume":"133","author":"Scrivano","year":"2019","journal-title":"Int. J. Leg. Med."},{"key":"ref_24","doi-asserted-by":"crossref","first-page":"69","DOI":"10.1186\/s41935-018-0102-6","article-title":"Estimation of the postmortem interval using GAPDH mRNA in skin and heart tissues of albino rats at different environmental conditions","volume":"8","author":"Elghamry","year":"2018","journal-title":"Egypt. J. Forensic Sci."},{"key":"ref_25","doi-asserted-by":"crossref","unstructured":"Chervoneva, I., Li, Y., Schulz, S., Croker, S., Wilson, C., Waldman, S.A., and Hyslop, T. (2010). Selection of optimal reference genes for normalization in quantitative RT-PCR. BMC Bioinform., 11.","DOI":"10.1186\/1471-2105-11-253"},{"key":"ref_26","doi-asserted-by":"crossref","unstructured":"Cianci, V., Mondello, C., Sapienza, D., Guerrera, M.C., Cianci, A., Crac\u00f2, A., Omero, F., Gioffr\u00e8, V., Gualniera, P., and Asmundo, A. (2024). Potential Role of mRNA in Estimating Postmortem Interval: A Systematic Review. Int. J. Mol. Sci., 25.","DOI":"10.3390\/ijms25158185"},{"key":"ref_27","doi-asserted-by":"crossref","first-page":"136","DOI":"10.1007\/s00414-006-0131-9","article-title":"Successful RNA extraction from various human postmortem tissues","volume":"121","author":"Heinrich","year":"2007","journal-title":"Int. J. Leg. Med."},{"key":"ref_28","doi-asserted-by":"crossref","first-page":"1547","DOI":"10.1007\/s00414-016-1349-9","article-title":"Postmortem muscle protein degradation in humans as a tool for PMI delimitation","volume":"130","author":"Pittner","year":"2016","journal-title":"Int. J. Leg. Med."},{"key":"ref_29","doi-asserted-by":"crossref","first-page":"1370","DOI":"10.1152\/japplphysiol.00343.2021","article-title":"Human adipose and skeletal muscle tissue DNA, RNA, and protein content","volume":"131","author":"Stroh","year":"2021","journal-title":"J. Appl. Physiol."}],"container-title":["International Journal of Molecular Sciences"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/www.mdpi.com\/1422-0067\/26\/15\/7059\/pdf","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2025,10,9]],"date-time":"2025-10-09T18:14:07Z","timestamp":1760033647000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.mdpi.com\/1422-0067\/26\/15\/7059"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2025,7,22]]},"references-count":29,"journal-issue":{"issue":"15","published-online":{"date-parts":[[2025,8]]}},"alternative-id":["ijms26157059"],"URL":"https:\/\/doi.org\/10.3390\/ijms26157059","relation":{},"ISSN":["1422-0067"],"issn-type":[{"value":"1422-0067","type":"electronic"}],"subject":[],"published":{"date-parts":[[2025,7,22]]}}}