Perikaryal accumulation and proteolysis of neurofilament proteins in the post-mortem rat brain.
Investigations of neurofilament alterations in neurodegenerative disorders utilize postmortem human tissues obtained at autopsy. To determine if alterations in the levels or distribution of neurofilament proteins might occur during the interval between death and autopsy, the postmortem cooling curve of the human brain was modeled in Sprague-Dawley rats and neurofilament proteins were examined by immunocytochemistry and immunoblots. One hour after death, enhanced perikaryal immunostaining of NF-M and both phosphorylated and nonphosphorylated NF-H epitopes was observed throughout the hippocampal formation. A greater number of neurons exhibited increased somatic immunostaining 4-h postmortem. In addition, loss of neurofilament protein immunostaining was observed in the neuropil, particularly in the molecular layer of the dentate gyrus. This corresponded with, but lagged behind, the pattern of calpain activation determined using an antibody against calpain-cleaved alpha-spectrin. Immunoblots confirmed the postmortem loss of neurofilament proteins in both triton-soluble and insoluble fractions. These results demonstrate that the levels and localization of neurofilament proteins observed in tissues obtained at autopsy even with short postmortem intervals may not accurately reflect the premortem condition.
Geddes
J W
JW
Sanders-Brown Center on Aging, University of Kentucky, Lexington 40536-0230, USA.
Bondada
V
V
Tekirian
T L
TL
Pang
Z
Z
Siman
R G
RG
D000704:Analysis of Variance; D000818:Animals; D001921:Brain / Q000201:enzymology / Q000378:metabolism*; D002154:Calpain / Q000378:metabolism; D004591:Electrophoresis, Polyacrylamide Gel; D004789:Enzyme Activation; D006624:Hippocampus / Q000378:metabolism; D015151:Immunoblotting; D007150:Immunohistochemistry; D008297:Male; D016900:Neurofilament Proteins / Q000378:metabolism*; D009474:Neurons / Q000201:enzymology / Q000378:metabolism; D010766:Phosphorylation; D011180:Postmortem Changes; D051381:Rats; D017207:Rats, Sprague-Dawley
D016900:Neurofilament Proteins; D002154:Calpain