Influence of post-mortem delay and storage temperature on the immunohistochemical detection of antigens in the CNS of mice

Abstract

The aim of this work was to compare the results of histochemical and immunohistochemical methods using mouse brains which were fixed with various post-mortem delays and storage temperatures (at a constant 4 degrees C or 22 degrees C, or at gradually decreasing post-mortem temperatures, mimicking conditions of human corpse). We studied the effects of post-mortem delay on glial fibrillary acidic protein, extracellular matrix components to which Wisteria floribunda agglutinin binds, non-phosphorylated neurofilament H, synaptophysin, calbindin and nitric oxide synthase isoenzymes. At the light microscopic level first signs of post-mortem changes were detectable after 6 h. Glial fibrillary acidic protein was most affected by post-mortem delay since its immunoreactivity increased dramatically with increasing post-mortem delay. N-acetylgalactosamines-beta1 labeled lectin binding sites, calbindin and intraneuronal non-phosphorylated neurofilament H seemed to be stable up to 12 h post-mortem. Storage temperature influenced the NADPH-d activity and the content of synaptophysin immunoreactivity to higher degree than all of the other parameters. We found only marginal differences of alterations comparing neocortex, hippocampus and corpus callosum. Our results indicate that different antigens are affected differently by the ongoing catabolic processes during post-mortem delay.