The effects of alpha-latrotoxin of black widow spider venom on synaptosome ultrastructure. A morphometric analysis...

The morphological effects of alpha-latrotoxin, the major component of black widow spider venom, were studied quantitatively in a crude synaptosome fraction (prepared from rat brain cortices) which was incubated at 37 degrees C for 10 min in Ringer solutions. Two toxin concentrations were employed, one causing a very large stimulation of transmitter release (approximately 65% and approximately 43% release of [3H]noradrenaline from preloaded synaptosomes, with and without Ca2+ in the incubation buffer), the other 50-60% as active. Incubated synaptosomes, fixed in suspension with aldehydes, were evenly dispersed in agarose before embedding, to assure randomized sampling in the subsequent morphometric analysis. In all the experimental conditions investigated, alpha-latrotoxin treatment caused a significant decrease in the density (number/unit area) of synaptic vesicles in synaptosome profiles. Such an effect was dose-dependent and partially Ca2+-dependent, in good agreement with the data on transmitter release. At high toxin concentration a moderate increase of synaptosome volume and surface area was observed, both with and without Ca2+. Mitochondrial swelling appeared only in synaptosomes treated in Ca2+ containing medium. These effects of alpha-latrotoxin are similar to those described previously at the neuromuscular junction. Thus, the toxin might be a tool of general use for studying vertebrate synapses.