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

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Abstract

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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Acknowledgments

The Brain and Body Donation Program is supported by the National Institute of Neurological Disorders and Stroke (U24 NS072026 National Brain and Tissue Resource for Parkinson’s Disease and Related Disorders), the National Institute on Aging (P30 AG19610 Arizona Alzheimer’s Disease Core Center), the Arizona Department of Health Services (contract 211002, Arizona Alzheimer’s Research Center), the Arizona Biomedical Research Commission (contracts 4001, 0011, 05-901 and 1001 to the Arizona Parkinson’s Disease Consortium) and the Michael J. Fox Foundation for Parkinson’s Research.

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  1. Douglas G. Walker & Lih-Fen Lue

    Present address: Laboratory of Neuroinflammation, Biodesign Neurodegenerative Disease Research Center, Arizona State University, Room 538, School of Life Sciences – E Wing, 427 E. Tyler Mall, Tempe, AZ, 85287, USA

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  1. Banner Sun Health Research Institute, Sun City, AZ, USA

    Douglas G. Walker, Alexis M. Whetzel, Geidy Serrano, Lucia I. Sue & Thomas G. Beach

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  1. Douglas G. Walker
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Walker, D.G., Whetzel, A.M., Serrano, G. et al. Characterization of RNA isolated from eighteen different human tissues: results from a rapid human autopsy program. Cell Tissue Bank 17, 361–375 (2016). https://doi.org/10.1007/s10561-016-9555-8

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