Studies on RNA integrity and gene expression in human myocardial tissue, pericardial fluid and blood, and its postmortem stability

González-Herrera, Lucas; Valenzuela, Abel; Marchal, Juan A.; Lorente, José A.; Villanueva, Enrique

doi:10.1016/j.forsciint.2013.08.001

Cited by 49 publications

(33 citation statements)

References 52 publications

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“…The factors that could affect the quality of tissue and tissue-derived products include disease conditions of the subject, agonal state of the subject immediately premortem, postmortem interval, pH of the tissue, tissue handling and preservation techniques, storage conditions and length of time in storage. Although the majority of studies of brain RNA have concluded that PMI does not have an overall significant effect on RNA stability over the time periods under consideration, the methods used to assess this have varied considerably (Auer et al 2014; Burke et al 1991; Condelli et al 2014; Fajardy et al 2009; Gonzalez-Herrera et al 2013; Hatzis et al 2011; Birdsill et al 2011; Burke et al 1991; Cummings et al 2001; Heinrich et al 2007; Koppelkamm et al 2011; Stan et al 2006). It is apparent that some mRNAs, for example cyclooxygenase-2 mRNA in brain (Lukiw and Bazan 1997), are lost quickly while others are more resistant (Nagy et al 2015; Birdsill et al 2011).…”

Section: Discussionmentioning

confidence: 99%

“…This study concluded, however, that heart RNA was significantly more stable than brain RNA. One other study concluded that heart RNA quality was not related to PMI (Gonzalez-Herrera et al 2013). …”

Section: Discussionmentioning

confidence: 99%

“…We also showed for heart that the raw gene expression values (Ct) for a range of housekeeping normalization genes could be affected by RIN. These genes are widely used to normalize for expression of genes of interest with the assumption that they remain relatively constant (Barber et al 2005; Dheda et al 2004; Gonzalez-Herrera et al 2013; Vandesompele et al 2002). A recent study reported that expression of common housekeeping genes is actually more stable with increased PMI than many other transcripts (Nagy et al 2015).…”

Section: Discussionmentioning

confidence: 99%

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Characterization of RNA isolated from eighteen different human tissues: results from a rapid human autopsy program

et al. 2016

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Many factors affect the integrity of messenger RNA from human autopsy tissues including postmortem interval (PMI) between death and tissue preservation and the pre-mortem agonal and disease states. In this communication, we describe RNA isolation and characterization of 389 samples from 18 different tissues from elderly donors who were participants in a rapid whole-body autopsy program located in Sun City, Arizona (www.brainandbodydonationprogram.org). Most tissues were collected within a PMI of 2–6 h (median 3.15 h; N = 455), but for this study, tissue from cases with longer PMIs (1.25–29.25 h) were included. RNA quality was assessed by RNA integrity number (RIN) and total yield (ng RNA/mg tissue). RIN correlated with PMI for heart (r = −0.531, p = 0.009) and liver (r = −558, p = 0.0017), while RNA yield correlated with PMI for colon (r = −485, p = 0.016) and skin (r = −0.460, p = 0.031). RNAs with the lowest integrity were from skin and cervix where 22.7 and 31.4 % of samples respectively failed to produce intact RNA; by contrast all samples from esophagus, lymph node, jejunum, lung, stomach, submandibular gland and kidney produced RNA with measurable RINs. Expression levels in heart RNA of 4 common housekeeping normalization genes showed significant correlations of Ct values with RIN, but only one gene, glyceraldehyde-3 phosphate dehydrogenase, showed a correlation of Ct with PMI. There were no correlations between RIN values obtained for liver, adrenal, cervix, esophagus and lymph node and those obtained from corresponding brain samples. We show that high quality RNA can be produced from most human autopsy tissues, though with significant differences between tissues and donors. The RNA stability and yield did not depend solely on PMI; other undetermined factors are involved, but these do not include the age of the donor.

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Section: Discussionmentioning

confidence: 99%

Section: Discussionmentioning

confidence: 99%

Section: Discussionmentioning

confidence: 99%

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Characterization of RNA isolated from eighteen different human tissues: results from a rapid human autopsy program

et al. 2016

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“…The reason why these tissues were chosen is that it is located deeply inside the body; thus, it is less affected by a change in external temperature. Some signaling protein components, glycogen-regulating enzymes, and housekeeping protein were selected as PMI protein candidates according to the previous studies of mRNA degradation profiles [6, 19]. Furthermore, a lateral flow assay (LFA)-based diagnostic chip was used to possibly determine the PMI using the recombinant rat GAPDH protein as a pilot attempt.…”

Section: Introductionmentioning

confidence: 99%

Degradation of Kidney and Psoas Muscle Proteins as Indicators of Post-Mortem Interval in a Rat Model, with Use of Lateral Flow Technology

et al. 2016

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We investigated potential protein markers of post-mortem interval (PMI) using rat kidney and psoas muscle. Tissue samples were taken at 12 h intervals for up to 96 h after death by suffocation. Expression levels of eight soluble proteins were analyzed by Western blotting. Degradation patterns of selected proteins were clearly divided into three groups: short-term, mid-term, and long-term PMI markers based on the half maximum intensity of intact protein expression. In kidney, glycogen synthase (GS) and glycogen synthase kinase-3β were degraded completely within 48 h making them short-term PMI markers. AMP-activated protein kinase α, caspase 3 and GS were short-term PMI markers in psoas muscle. Glyceraldehyde 3-phosphate dehydrogenase (GAPDH) was a mid-term PMI marker in both tissues. Expression levels of the typical long-term PMI markers, p53 and β-catenin, were constant for at least 96 h post-mortem in both tissues. The degradation patterns of GS and caspase-3 were verified by immunohistochemistry in both tissues. GAPDH was chosen as a test PMI protein to perform a lateral flow assay (LFA). The presence of recombinant GAPDH was clearly detected in LFA and quantified in a concentration-dependent manner. These results suggest that LFA might be used to estimate PMI at a crime scene.

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“…On the human autopsy tissue, inevitably affected by post-mortem degradation, the messenger RNA (mRNA) expression yielded results that allowed qualitative and quantitative determination of gene expression in the various tissues of the body [2,10,12,21,24]. Among those, the cardiac tissue has been preferentially studied because of its involvement in the diagnosis of sudden cardiac death and a higher resistance to autolysis like any muscle tissue [11,19,22].…”

Section: Introductionmentioning

confidence: 99%

Morphological characteristics of the sinus node on postmortem tissue

Perde¹,

Atkinson

Yanni

et al. 2016

Folia Morphol

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Studies on RNA integrity and gene expression in human myocardial tissue, pericardial fluid and blood, and its postmortem stability

Cited by 49 publications

References 52 publications

Characterization of RNA isolated from eighteen different human tissues: results from a rapid human autopsy program

Characterization of RNA isolated from eighteen different human tissues: results from a rapid human autopsy program

Degradation of Kidney and Psoas Muscle Proteins as Indicators of Post-Mortem Interval in a Rat Model, with Use of Lateral Flow Technology

Morphological characteristics of the sinus node on postmortem tissue

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