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Erschienen in: International Journal of Legal Medicine 2/2007

01.03.2007 | Original Article

Successful RNA extraction from various human postmortem tissues

verfasst von: Marielle Heinrich, Katja Matt, Sabine Lutz-Bonengel, Ulrike Schmidt

Erschienen in: International Journal of Legal Medicine | Ausgabe 2/2007

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Abstract

Recently, several authors described the observation that RNA degradation does not correlate with the postmortem interval (PMI), but rather with other parameters like environmental impact and the circumstances of death. Therefore, the question arose if the analysis of gene expression could be a valuable tool in forensic genetics to contribute to the determination of the cause of death. In our study, six human tissues obtained from six individuals with PMI varying between 15 and 118 h were used for total RNA extraction. Quantification was performed using a GAPDH real-time assay, and the quality of mRNA was checked by amplification of different fragment lengths of the GAPDH transcript. In our set of samples, nearly all tissues in all PMI revealed satisfactory results, while skeletal muscle, followed by brain and heart, gave the best results. No correlation between PMI and RNA degradation could be detected, as very good results were observed for all tissues from the individual with the longest PMI. The highly promising results obtained in this study raise hopes that in the near future several fields of forensic investigation may profit from additional information about gene expression patterns and their correlation with pathological findings.
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Literatur
1.
Zurück zum Zitat Wilusz CJ, Wormington M, Peltz SW (2001) The cap-to-tail guide to mRNA turnover. Nat Rev Mol Cell Biol 2:237–246PubMedCrossRef Wilusz CJ, Wormington M, Peltz SW (2001) The cap-to-tail guide to mRNA turnover. Nat Rev Mol Cell Biol 2:237–246PubMedCrossRef
2.
Zurück zum Zitat Catts VS, Catts SV, Fernandez HR, Taylor JM, Coulson EJ, Lutze-Mann LH (2005) A microarray study of post-mortem mRNA degradation in mouse brain tissue. Brain Res Mol Brain Res 138:164–177PubMedCrossRef Catts VS, Catts SV, Fernandez HR, Taylor JM, Coulson EJ, Lutze-Mann LH (2005) A microarray study of post-mortem mRNA degradation in mouse brain tissue. Brain Res Mol Brain Res 138:164–177PubMedCrossRef
3.
Zurück zum Zitat Preece P, Cairns NJ (2003) Quantifying mRNA in postmortem human brain: influence of gender, age at death, postmortem interval, brain pH, agonal state and inter-lobe mRNA variance. Brain Res Mol Brain Res 118:60–71PubMedCrossRef Preece P, Cairns NJ (2003) Quantifying mRNA in postmortem human brain: influence of gender, age at death, postmortem interval, brain pH, agonal state and inter-lobe mRNA variance. Brain Res Mol Brain Res 118:60–71PubMedCrossRef
4.
Zurück zum Zitat Fitzpatrick R, Casey OM, Morris D et al (2002) Postmortem stability of RNA isolated from bovine reproductive tissues. Biochim Biophys Acta 1574:10–14PubMed Fitzpatrick R, Casey OM, Morris D et al (2002) Postmortem stability of RNA isolated from bovine reproductive tissues. Biochim Biophys Acta 1574:10–14PubMed
5.
Zurück zum Zitat Kobayashi H, Sakimura K, Kuwano R et al (1990) Stability of messenger RNA in postmortem human brains and construction of human brain cDNA libraries. J Mol Neurosci 2:29–34PubMed Kobayashi H, Sakimura K, Kuwano R et al (1990) Stability of messenger RNA in postmortem human brains and construction of human brain cDNA libraries. J Mol Neurosci 2:29–34PubMed
6.
Zurück zum Zitat Reddy PH, McWeeney S, Park BS et al (2004) Gene expression profiles of transcripts in amyloid precursor protein transgenic mice: upregulation of mitochondrial metabolism and apoptotic genes is an early cellular change in Alzheimer’s disease. Hum Mol Genet 13:1225–1240PubMedCrossRef Reddy PH, McWeeney S, Park BS et al (2004) Gene expression profiles of transcripts in amyloid precursor protein transgenic mice: upregulation of mitochondrial metabolism and apoptotic genes is an early cellular change in Alzheimer’s disease. Hum Mol Genet 13:1225–1240PubMedCrossRef
7.
Zurück zum Zitat Preece P, Virley DJ, Costandi M et al (2003) An optimistic view for quantifying mRNA in post-mortem human brain. Brain Res Mol Brain Res 116:7–16PubMedCrossRef Preece P, Virley DJ, Costandi M et al (2003) An optimistic view for quantifying mRNA in post-mortem human brain. Brain Res Mol Brain Res 116:7–16PubMedCrossRef
8.
Zurück zum Zitat Bauer M, Polzin S, Patzelt D (2003) Quantification of RNA degradation by semi-quantitative duplex and competitive RT-PCR: a possible indicator of the age of bloodstains? Forensic Sci Int 138:94–103PubMedCrossRef Bauer M, Polzin S, Patzelt D (2003) Quantification of RNA degradation by semi-quantitative duplex and competitive RT-PCR: a possible indicator of the age of bloodstains? Forensic Sci Int 138:94–103PubMedCrossRef
9.
Zurück zum Zitat Bauer M, Gramlich I, Polzin S, Patzelt D (2003) Quantification of mRNA degradation as possible indicator of postmortem interval—a pilot study. Leg Med (Tokyo) 5:220–227 Bauer M, Gramlich I, Polzin S, Patzelt D (2003) Quantification of mRNA degradation as possible indicator of postmortem interval—a pilot study. Leg Med (Tokyo) 5:220–227
10.
Zurück zum Zitat Bauer M, Patzelt D (2003) A method for simultaneous RNA and DNA isolation from dried blood and semen stains. Forensic Sci Int 136:76–78PubMedCrossRef Bauer M, Patzelt D (2003) A method for simultaneous RNA and DNA isolation from dried blood and semen stains. Forensic Sci Int 136:76–78PubMedCrossRef
11.
Zurück zum Zitat Loddenkötter B, Becker K, Hohoff C, Brinkmann B, Bajanowski T (2005) Real-time quantitative PCR assay for the detection of Helicobacter pylori: no association with sudden infant death syndrome. Int J Legal Med 119:202–206PubMedCrossRef Loddenkötter B, Becker K, Hohoff C, Brinkmann B, Bajanowski T (2005) Real-time quantitative PCR assay for the detection of Helicobacter pylori: no association with sudden infant death syndrome. Int J Legal Med 119:202–206PubMedCrossRef
12.
Zurück zum Zitat Bustin SA (2002) Quantification of mRNA using real-time reverse transcription PCR (RT-PCR): trends and problems. J Mol Endocrinol 29:23–39PubMedCrossRef Bustin SA (2002) Quantification of mRNA using real-time reverse transcription PCR (RT-PCR): trends and problems. J Mol Endocrinol 29:23–39PubMedCrossRef
13.
Zurück zum Zitat Bustin SA (2000) Absolute quantification of mRNA using real-time reverse transcription polymerase chain reaction assays. J Mol Endocrinol 25:169–193PubMedCrossRef Bustin SA (2000) Absolute quantification of mRNA using real-time reverse transcription polymerase chain reaction assays. J Mol Endocrinol 25:169–193PubMedCrossRef
14.
Zurück zum Zitat Goidin D, Mamessier A, Staquet MJ, Schmitt D, Berthier-Vergnes O (2001) Ribosomal 18S RNA prevails over glyceraldehyde-3-phosphate dehydrogenase and beta-actin genes as internal standard for quantitative comparison of mRNA levels in invasive and noninvasive human melanoma cell subpopulations. Anal Biochem 295:17–21PubMedCrossRef Goidin D, Mamessier A, Staquet MJ, Schmitt D, Berthier-Vergnes O (2001) Ribosomal 18S RNA prevails over glyceraldehyde-3-phosphate dehydrogenase and beta-actin genes as internal standard for quantitative comparison of mRNA levels in invasive and noninvasive human melanoma cell subpopulations. Anal Biochem 295:17–21PubMedCrossRef
15.
Zurück zum Zitat Aerts JL, Gonzales MI, Topalian SL (2004) Selection of appropriate control genes to assess expression of tumor antigens using real-time RT-PCR. Biotechniques 36:84–86, 88, 90–91PubMed Aerts JL, Gonzales MI, Topalian SL (2004) Selection of appropriate control genes to assess expression of tumor antigens using real-time RT-PCR. Biotechniques 36:84–86, 88, 90–91PubMed
16.
Zurück zum Zitat Phang TW, Shi CY, Chia JN, Ong CN (1994) Amplification of cDNA via RT-PCR using RNA extracted from postmortem tissues. J Forensic Sci 39:1275–1279PubMed Phang TW, Shi CY, Chia JN, Ong CN (1994) Amplification of cDNA via RT-PCR using RNA extracted from postmortem tissues. J Forensic Sci 39:1275–1279PubMed
17.
Zurück zum Zitat Mall G, Eckl M, Sinicina I, Peschel O, Hubig M (2005) Temperature-based death time estimation with only partially known environmental conditions. Int J Legal Med 119:185–194PubMedCrossRef Mall G, Eckl M, Sinicina I, Peschel O, Hubig M (2005) Temperature-based death time estimation with only partially known environmental conditions. Int J Legal Med 119:185–194PubMedCrossRef
18.
Zurück zum Zitat Henssge C, Madea B (2004) Estimation of the time since death in the early post-mortem period. Forensic Sci Int 144:167–175PubMedCrossRef Henssge C, Madea B (2004) Estimation of the time since death in the early post-mortem period. Forensic Sci Int 144:167–175PubMedCrossRef
19.
Zurück zum Zitat Takamiya M, Saigusa K, Kumagai R, Nakayashiki N, Aoki Y (2005) Studies on mRNA expression of tissue-type plasminogen activator in bruises for wound age estimation. Int J Legal Med 119:16–21PubMedCrossRef Takamiya M, Saigusa K, Kumagai R, Nakayashiki N, Aoki Y (2005) Studies on mRNA expression of tissue-type plasminogen activator in bruises for wound age estimation. Int J Legal Med 119:16–21PubMedCrossRef
20.
Zurück zum Zitat Hayashi T, Ishida Y, Kimura A, Takayasu T, Eisenmenger W, Kondo T (2004) Forensic application of VEGF expression to skin wound age determination. Int J Legal Med 118:320–325PubMedCrossRef Hayashi T, Ishida Y, Kimura A, Takayasu T, Eisenmenger W, Kondo T (2004) Forensic application of VEGF expression to skin wound age determination. Int J Legal Med 118:320–325PubMedCrossRef
21.
Zurück zum Zitat Suay L, Salvador ML, Abesha E, Klein U (2005) Specific roles of 5′ RNA secondary structures in stabilizing transcripts in chloroplasts. Nucleic Acids Res 33:4754–4761PubMedCrossRef Suay L, Salvador ML, Abesha E, Klein U (2005) Specific roles of 5′ RNA secondary structures in stabilizing transcripts in chloroplasts. Nucleic Acids Res 33:4754–4761PubMedCrossRef
22.
Zurück zum Zitat Ikematsu K, Tsuda R, Nakasono I (2006) Gene response of mouse skin to pressure injury in the neck region. Leg Med (Tokyo) 8:128–131 Ikematsu K, Tsuda R, Nakasono I (2006) Gene response of mouse skin to pressure injury in the neck region. Leg Med (Tokyo) 8:128–131
23.
Zurück zum Zitat Liu J, Lewohl JM, Dodd PR, Randall PK, Harris RA, Mayfield RD (2004) Gene expression profiling of individual cases reveals consistent transcriptional changes in alcoholic human brain. J Neurochem 90:1050–1058PubMedCrossRef Liu J, Lewohl JM, Dodd PR, Randall PK, Harris RA, Mayfield RD (2004) Gene expression profiling of individual cases reveals consistent transcriptional changes in alcoholic human brain. J Neurochem 90:1050–1058PubMedCrossRef
Metadaten
Titel
Successful RNA extraction from various human postmortem tissues
verfasst von
Marielle Heinrich
Katja Matt
Sabine Lutz-Bonengel
Ulrike Schmidt
Publikationsdatum
01.03.2007
Verlag
Springer-Verlag
Erschienen in
International Journal of Legal Medicine / Ausgabe 2/2007
Print ISSN: 0937-9827
Elektronische ISSN: 1437-1596
DOI
https://doi.org/10.1007/s00414-006-0131-9

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