Quantifying synaptic number and structure: effects of stain and post-mortem delay.
Current research indicates the importance of synaptic number and structure in plastic processes such as development, learning and memory, and aging. As such, the examination of these neural features has become an important factor in research on human conditions such as mental retardation, aging and Alzheimer's disease. Synaptic research in human tissue typically involves delayed post-mortem fixation, therefore the current research was designed to examine the effect of post-mortem delay on synaptic number and structure in tissue stained with either routine osmium lead citrate/uranyl acetate (osmium) or ethanol phosphotungstic acid (EPTA). Results indicate that synaptic density shows either a gradual decline (EPTA) or an initial marked drop followed by a plateau (osmium) up to 10-15 h post-mortem depending on the stain used. The number of synaptic vesicles per synapse also undergoes a gradual decline. Measures of synaptic structure were more stable, with the primary change being an initial increase in the cross-sectional length of the synapse. Maximal height of the pre- and postsynaptic dense elements were not affected by post-mortem delay. The EPTA stain gave the best estimates of synaptic parameters with short post-mortem delays. These results indicate that different synaptic measures (and stains) show different responses to post-mortem fixation delay, and that experimental or statistical methods must be used to control for post-mortem effects.
Department of Psychology, University of Toronto, Scarborough, Ont., Canada.
T L
TL
Petit
J C
JC
LeBoutillier
D000818:Animals; D002452:Cell Count; D002540:Cerebral Cortex / Q000187:drug effects / Q000648:ultrastructure*; D008297:Male; D008854:Microscopy, Electron / Q000379:methods*; D009942:Organometallic Compounds; D009992:Osmium; D009993:Osmium Tetroxide; D010772:Phosphotungstic Acid; D011180:Postmortem Changes; D051381:Rats; D013194:Staining and Labeling; D013569:Synapses / Q000187:drug effects / Q000648:ultrastructure*
D009942:Organometallic Compounds; D010772:Phosphotungstic Acid; C005460:uranyl acetate; D009992:Osmium; D009993:Osmium Tetroxide