Presynaptic ecto- and postsynaptic endo-calcium-adenosine-triphosphatases in synaptosomes: doubts about biochemical...

Ecto- and endo-Ca-adenosine-triphosphatase (ATPase) activity was identified as electron-dense lead or cerium phosphate precipitate in the rat cortical synaptosomes by transmission electron microscopy and enzyme histochemistry. The formation of the deposit was dependent on the presence of ATP (the substrate), Ca (activator) and levamisole, quercetin or ouabain (inhibitors of different phosphatases and ATPases). Reaction products were found at the external surface of the presynaptic membrane, both surfaces of the postsynaptic membrane, in the synaptic cleft and in the free mitochondrial membranes. In the presence of ATP and the three inhibitors together, the quantity of the precipitate decreased markedly, but we still found some deposit on the external surface of the presynaptic membrane (this activity is probably due to the so-called ecto-ATPase) and on the internal surface of the postsynaptic one (endo-ATPase). The distinction between ecto- and endo-ATPases in biochemical fractions solely upon biochemical differential measurements must be interpreted with caution.