High-resolution Quantification of Specific mRNA Levels in Human Brain Autopsies and Biopsies

Castensson, Anja; Emilsson, Lina; Preece, Paul; Jazin, Elena

doi:10.1101/gr.10.8.1219

Cited by 71 publications

(54 citation statements)

References 60 publications

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“…While the effects of PMI on global mRNA integrity are minimal, there may be transcript-specific degradation (Bahn et al, 2001;Barton et al, 1993;Castensson et al, 2000;Ervin et al, 2007;Gilmore et al, 1993;Harrison et al, 1995;Leonard et al, 1993;OniOrisan et al, 2008;Pardue et al, 1994;Stan et al, 2006;Trotter et al, 2002). Several studies have specifically examined the effects of PMI on protein measures in the brain.…”

Section: Postmortem Interval (Pmi)mentioning

confidence: 99%

Postmortem Brain: An Underutilized Substrate for Studying Severe Mental Illness

McCullumsmith

Hammond

Shan

et al. 2013

Neuropsychopharmacol

106

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We propose that postmortem tissue is an underutilized substrate that may be used to translate genetic and/or preclinical studies, particularly for neuropsychiatric illnesses with complex etiologies. Postmortem brain tissues from subjects with schizophrenia have been extensively studied, and thus serve as a useful vehicle for illustrating the challenges associated with this biological substrate. Schizophrenia is likely caused by a combination of genetic risk and environmental factors that combine to create a disease phenotype that is typically not apparent until late adolescence. The complexity of this illness creates challenges for hypothesis testing aimed at understanding the pathophysiology of the illness, as postmortem brain tissues collected from individuals with schizophrenia reflect neuroplastic changes from a lifetime of severe mental illness, as well as treatment with antipsychotic medications. While there are significant challenges with studying postmortem brain, such as the postmortem interval, it confers a translational element that is difficult to recapitulate in animal models. On the other hand, data derived from animal models typically provide specific mechanistic and behavioral measures that cannot be generated using human subjects. Convergence of these two approaches has led to important insights for understanding molecular deficits and their causes in this illness. In this review, we discuss the problem of schizophrenia, review the common challenges related to postmortem studies, discuss the application of biochemical approaches to this substrate, and present examples of postmortem schizophrenia studies that illustrate the role of the postmortem approach for generating important new leads for understanding the pathophysiology of severe mental illness.

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Section: Postmortem Interval (Pmi)mentioning

confidence: 99%

Postmortem Brain: An Underutilized Substrate for Studying Severe Mental Illness

McCullumsmith

Hammond

Shan

et al. 2013

Neuropsychopharmacol

106

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“…Although yields of RNA vary considerably, there is no correlation between the concentrations of specific transcripts and PMI (Johnson et al 1986;Lewohl et al 1997a;Hynd et al 2001). Only CYP26A1 has been reported to show a significant decrease in the mRNA level, and that is in samples taken after prolonged autopsy times (Castensson et al 2002). Thus, autopsy data should conform to relative mRNA transcript levels in vivo.…”

Section: Post-mortem Intervalmentioning

confidence: 99%

“…Nor does PMI appear to significantly affect the recovery of mRNA (typically 3-5% of total RNA), as judged by cDNA analysis (Johnson et al 1986;Perrett et al 1988;Burke et al 1991;Harrison et al 1991;Peltz et al 1991;Yasojima et al 2001;Bahn et al 2002;Castensson et al 2002). Despite slight degrada tion of cortical RNA, neither total RNA nor oligo-dTseparated mRNAs changed for up to 36 h post-mortem (Johnson et al 1988).…”

Section: Post-mortem Intervalmentioning

confidence: 99%

“…Similarly, although RNA is highly labile in vivo, it can routinely be extracted from frozen autopsy tissue (Johnson et al 1986;Wood et al 1986;Ross et al 1992;Barton et al 1993;Leonard et al 1993;Johnston et al 1997;Schramm et al 1999;Castensson et al 2000;Zhou et al 2001a), rapidly procured non-frozen autopsy tissue (Cummings et al 2001) and formalin-fixed, paraffin-embedded autopsy tissue (Godfrey et al 2000;Hawkins and Dodd 2000). The established method for assessing RNA quality and quantity is spectrophotometric measurement of the 260 : 280 nm absorbance ratio (Glasel 1995).…”

Section: Molecular Integritymentioning

confidence: 99%

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Biochemical and molecular studies using human autopsy brain tissue

Hynd

Lewohl

Scott

et al. 2003

Journal of Neurochemistry

221

171

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The use of human brain tissue obtained at autopsy for neurochemical, pharmacological and physiological analyses is reviewed. RNA and protein samples have been found suitable for expression profiling by techniques that include RT-PCR, cDNA microarrays, western blotting, immunohistochemistry and proteomics. The rapid development of molecular biological techniques has increased the impetus for this work to be applied to studies of brain disease. It has been shown that most nucleic acids and proteins are reasonably stable postmortem. However, their abundance and integrity can exhibit marked intra-and intercase variability, making comparisons between case-groups difficult. Variability can reveal important functional and biochemical information. The correct interpretation of neurochemical data must take into account such factors as age, gender, ethnicity, medicative history, immediate ante-mortem status, agonal state and post-mortem and post-autopsy intervals. Here we consider issues associated with the sampling of DNA, RNA and proteins using human autopsy brain tissue in relation to various ante-and postmortem factors. We conclude that valid and practical measures of a variety of parameters may be made in human brain tissue, provided that specific factors are controlled.

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“…Increasingly, human postmortem brain tissue is being used to quantify cellular and molecular markers of neural processes in the pursuit of understanding normal brain function and correlates of dysfunction in CNS illnesses (Casanova and Kleinman, 1990;Castensson et al, 2000;Harrison et al, 1994;Mirnics et al, 2001;Ravid et al, 1992;Torrey et al, 2000). Because of the inaccessibility of the human brain and its unavailability for biopsy, fresh tissue is rarely obtainable.…”

Section: Introductionmentioning

confidence: 99%

Human postmortem tissue: What quality markers matter?

Ghose²,

et al. 2006

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Post mortem human brain tissue is used for the study of many different brain diseases. A key factor in conducting postmortem research is the quality of the tissue. Unlike animal tissue, whose condition at death can be controlled and influenced, human tissue can only be collected naturalistically. This introduces potential confounds, based both on pre-and postmortem conditions, that may influence the quality of tissue and its ability to yield accurate results. The traditionally recognized confounds that reduce tissue quality are agonal factors (e.g., coma, hypoxia, hyperpyrexia at the time of death), and long postmortem interval (PMI). We measured tissue quality parameters in over 100 postmortem cases collected from different sources and correlated them with RNA quality (as indicated by the RNA Integrity Number (RIN)) and with protein quality (as measured by the level of representative proteins). Our results show that the most sensible indicator of tissue quality is RIN and that there is a good correlation between RIN and the pH. No correlation developed between protein levels and the aforementioned factors. Moreover, even when RNA was degraded, the protein levels remained stable. However, these correlations did not prove true under all circumstances (e.g. thawed tissue, surgical tissue), that yielded unexpected quality indicators. These data also suggest that cases whose source was a Medical Examiner's office represent high tissue quality.

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High-resolution Quantification of Specific mRNA Levels in Human Brain Autopsies and Biopsies

Cited by 71 publications

References 60 publications

Postmortem Brain: An Underutilized Substrate for Studying Severe Mental Illness

Postmortem Brain: An Underutilized Substrate for Studying Severe Mental Illness

Biochemical and molecular studies using human autopsy brain tissue

Human postmortem tissue: What quality markers matter?

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