Effects of freezing storage time on the density of muscarinic receptors in the human postmortem brain: an autoradiographic study in control and Alzheimer's disease brain tissues

Abstract

The effect of sex, age (range = 41–84 years), postmortem delay (range = 1–71 h) and freezing storage time (FST) (range = 8–75 months) at −25°C on the density of muscarinic receptors (MR) was examined in tissue sections of several representative areas of 41 postmortem brains from adult patients who had died from non-neurological disorders using [ 3H] N-methylscopolamine as a ligand. Neither age, sex nor postmortem delay determined significant changes in the density of MR in frontal and entorhinal cortex, hippocampus and striatum. By contrast, FST significantly decreased the densities of MR in frontal and entorhinal cortex, pyramidal layer of CA1 and CA3 fields at the hippocampus and over caudate nucleus. This reduction in MR densities did not reach statistical significance, for any region, when FST was less than 39 months. Although there was a tendency towards a decrease, no significant changes were observed in putamen and over hippocampal dentate gyrus. FST (range = 11–78 months) also significantly decreased the densities of MR in the same regions of postmortem brains from 18 patients who had died with a clinico-pathological diagnosis of Alzheimer's disease (AD). Even though there was a general tendency towards a decrease (between 7% in the caudate and 30% in the dentate gyrus at the hippocampus), no significant differences could be seen in MR densities between control and AD cases, except in the hilus in the dentate gyrus ( P < 0.022), when brains were matched for FST. From the present results it is clear that control and diseased brains must also be matched for FST as well as for other factors such as sex, age and postmortem delay. It is possible that differences in FST could in part account for the variability of the reported results measuring MR in control and AD brains. At least for MR, FST shorter than three years would seem to be acceptable when performing this kind of studies.