Recovery and expression of messenger RNA from postmortem human brain tissue

The Bryan Alzheimer's Disease Research Center Rapid Autopsy Program at Duke University Medical Center obtains postmortem human brain tissue for experi mental investigations. We evaluated 19 brains for RNA integrity and mRNA ge ne expression. Nine were from patients diagnosed with Alzheimer's disease, and ten were from nondemented controls. In all cases, the following variabl es were recorded: postmortem procurement delay (range, 1 hour and 10 minute s to 14 hours), pH of cerebrospinal fluid, premortem fever or sepsis, provi sion of supplemental oxygen in the agonal period, and temporal relation to time of death (either sudden death or protracted illness). Total RNA was ex tracted, quantified, and evaluated by agarose gel electrophoresis and quant itative gene expression analysis of 18S rRNA and edg-1 using TaqMan technol ogy. All samples appeared to yield intact RNA without significant degradati on, and expression of the edg-1 gene was detected by the real time reverse transcriptase polymerase chain reaction in all cases. We conclude that inta ct RNA can be obtained from postmortem human brain tissue, even in patients with severe premortem illnesses and delayed postmortem tissue procurement intervals. However, we caution that the successful expression of certain ge nes from postmortem brain tissue may require enhanced procurement efforts t o maximize RNA integrity.