Relative postmortem stability of spinal motoneuronal proteins detectable by two-dimensional electrophoresis

The suitability of using spinal tissue several hours after death for analysis by high resolution two-dimensional electrophoresis has been examined. It was found that many of the proteins of bovine spinal motoneurons detectable on two-dimensional polyacrylamide gels appear to be relatively stable in situ at room temperature during the first postmortem day. When extracts of total proteins from ventral roots and motoneuronal cell bodies isolated from 1-d-old tissue were examined, all spots could be matched to control gels. Upon visual inspection of the gels, postmortem changes in the amount of stain associated with a spot were obvious in three of 364 proteins from isolated motoneuronal cell bodies and none of 237 proteins from ventral roots. Other proteins underwent quantitative changes that were detected only after computer-assisted densitometry on the gels, whereas some did not appear to change at all. In the neuropil surrounding the motoneuron cell bodies, more pronounced changes in protein patterns occurred during the postmortem period. We conclude that properly controlled two-dimensional electrophoretic analyses of postmortem spinal tissue can provide reliable qualitative and quantitative information about the antemortem protein composition of spinal motoneurons.