Rapid autopsy brains for biochemical research: Experiences in establishing a programme

Procter, Andrew W.; Doshi, Bala; Bowen, David M.; Murphy, Elaine

doi:10.1002/gps.930050504

Cited by 18 publications

(13 citation statements)

References 33 publications

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“…Uneven regional distribution of nucleotide metabolism has also been observed (Kovacs et al 1998). To ensure reproducibility, consistent dissection procedures must be followed (Procter et al 1990). …”

Section: Tissue Dissectionmentioning

confidence: 99%

Biochemical and molecular studies using human autopsy brain tissue

Hynd

Lewohl

Scott

et al. 2003

Journal of Neurochemistry

222

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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Section: Tissue Dissectionmentioning

confidence: 99%

Biochemical and molecular studies using human autopsy brain tissue

Hynd

Lewohl

Scott

et al. 2003

Journal of Neurochemistry

222

171

View full text Add to dashboard Cite

show abstract

“…Thus, perhaps the best way to ascertain the influence of AS on RNA is the use of brains from patients whose premortem course was prospectively assessed during their terminal illness. This has been accomplished for a series of dementia patients (Procter et al, 1990). Using tissue from this series, the amount of m l muscarinic receptor mRNA in temporal cortex was found to be influenced by terminal coma; that is, a significant correlation was observed between increasing duration of coma (1-336 h) and decreasing hybridization signal (Harrison et al,199 la).…”

Section: Premortem Influences On Human Brain Rnamentioning

confidence: 99%

Pre‐and Postmortem Influences on Brain RNA

Barton

Pearson

Najlerahim

et al. 1993

Journal of Neurochemistry

243

142

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Many potentially valuable techniques for the understanding of human neurobiological and neuropathological processes require the use of RNA obtained from postmortem tissue. As with earlier neurochemical studies, there are two particular problems posed by such tissue in comparison with tissue from experimental animals. These are the postmortem interval and the condition of the patient prior to death, referred to as the agonal state. We review the nature and extent of the effects of postmortem interval and agonal state on RNA in brain tissue, with particular reference to the study of neuropsychiatric disorders. Perhaps surprisingly, postmortem interval has at most a modest effect on RNA. Abundant intact and biologically active RNA is present in tissue frozen 36 h or more after death. Postmortem interval does not account for the marked variability observed among human brains in all RNA parameters. Despite the overall stability of RNA after death, some evidence suggests that individual RNAs may undergo postmortem decay. Less attention has been paid to the effects of agonal state. The existing data indicate that events in the premortem period such as hypoxia and coma can affect the amount of some messenger RNAs. The nature of agonal state influences depends on the messenger RNA in question, though the basis for this selective vulnerability is unknown. No agonal state effect on overall RNA level or activity has been found. The data show that postmortem brain tissue can be used for RNA research. However, considerable attention must be paid to controlling for the influences of pre-and postmortem factors, especially when quantitative analyses are performed.

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“…This rigorous standard for comparison made it virtually impossible to obtain brains of normal subjects matched to the AD cases for these fac tors. Therefore, the group of non-Alzheimer dementias represent a more appropriate control [ 15], albeit of sever al quite different disease entities.…”

Section: Discussionmentioning

confidence: 99%

“…The remaining cases were one case each of Pick's disease, vascular dementia, pseudodementia and multisys tem degeneration, as described in Lowe et al [14]. One hemisphere had been fixed in 10% neutral formalin for full ncuropathological examination and the other sliced coronally into I -cm thick slices and placed into an ice-cold Krcbs-Ringer phosphate buffer containing glucose and frozen (-70°C) within 4 h of death [15], The routincpm-dclay brains were from both AD subjects (6 females, 2 males) and controls (4 females, 4 males). The AD group had a mean age of 80 ± 5 years (controls: 77 ± 12 years) and pm delay of 38 ± 15 h (40 ± 11).…”

Section: Brain Materialsmentioning

confidence: 99%

Brain Membrane Serine Protease Activity in Human Cortex Compared with Rat: Implication for Alzheimer's Disease

Stratmann

Webster²,

Francis³

et al. 1994

Dement Geriatr Cogn Disord

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Cerebral cortex from humans and rats was extracted sequentially with detergent-containing and low-ionic-strength buffers. The resulting pellet was extracted with detergent/high-ionic-strength buffer to yield a soluble enzyme preparation. This was incubated with substrate prepared from rat cerebral cortical membranes containing amyloid precursor protein-like immunoreactivity (APPLIR) of 116 kD approximate apparent molecular mass. The effectiveness of various enzyme preparations to degrade APPLIR was: routine-post-mortem (pm)-delay human samples > rat pup > short-pm-delay human samples >> adult rat. In incubations with human samples only a 100-kD product accumulated. The activity in human brain was inhibited by phenylmethylsulphonylfluoride, insensitive to Ca²⁺, correlated with pyramidal neurone numbers but not those of astrocytes and was not significantly higher in Alzheimer''s disease compared with controls. These data are discussed in terms of other approaches for studying proteolytic activity to explain the deposition of β-amyloid protein in this disease.

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Rapid autopsy brains for biochemical research: Experiences in establishing a programme

Cited by 18 publications

References 33 publications

Biochemical and molecular studies using human autopsy brain tissue

Biochemical and molecular studies using human autopsy brain tissue

Pre‐and Postmortem Influences on Brain RNA

Brain Membrane Serine Protease Activity in Human Cortex Compared with Rat: Implication for Alzheimer's Disease

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