A STUDY ON THE OGAWA'S SYSTEM FOR CYTOCHEMICAL DEMONSTRATION OF OXIDOREDUCTASES WITH FERRICYANIDE AS ELECTRON ACCEPTOR

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Spectrophotometric study on kinetics of the reaction under the conditions mimicking its course in the tissue, carried out with α-glycerophosphate as substrate, rat spermatogenic mitochondria as a source of enzyme, tris/hydroxymethyl/aminomethane as copper complexing agent, and in pH 8.4, i. e. in such a range in which the final product, copper ferrocyanide, is soluble led to the following conclusions. Enzymatic oxidation of the substrate is immediately followed by ferricyanide reduction, whereas the final product preciptation starts with a few minutes latency. Dimethylsulfoxide (DMSO) in the final concentration of 5% does not alter the reaction course. However, at its concentration of 15% there is no latency in the final product formation that runs parallelly to the course of the reduction of ferricyanide but there is an inhibition of both the enzymatic reduction of ferricyanide and the formation of copper ferrocyanide as final reaction product. On fine structural level it was found that already at 5 per cent concentration of DMSO and at a shortened time of incubation the localization of the reaction for a-glycerophosphate: ferricyanide oxidoreductase in mitochondria and the other structures of rat spermatids was improved.