Quantitative dehydrogenase histochemistry with exogenous electron carriers (PMS, MPMS, MB).
The relative efficiencies of phenazine methosulfate (PMS), 1-methoxy-phenazine methosulfate (MPMS) and Meldola Blue (MB) as electron carriers were determined biochemically (non-enzymic NADH-tetrazolium salt-test) and by quantitative histochemistry (heart and kidney slices; succinate dehydrogenase, SDH; lactate dehydrogenase, LDH). MPMS developed the highest electron transfer velocity in biochemical assays. The reaction was independent of the pH value between 7.0-8.5. PMS and MB always showed a lower transfer ability in biochemical tests which was higher with iodonitrotetrazolium chloride (INT) than with nitro blue tetrazolium chloride (NBT). A distinct pH dependence was demonstrable with MB in this respect, preferentially using INT as tetrazolium salt. Quantitative histochemical results with electron carriers are often at variance with biochemical ones. MPMS leads to somewhat higher demonstrable activities only in the determination of the NAD-dependent LDH, whereas MB results in somewhat higher LDH activity than PMS (reaction medium with agarose). MB and PMS yielded almost equally high activities in the demonstration of the flavoprotein-dependent SDH using a reaction medium with agarose. With an aqueous reaction medium, PMS resulted in higher SDH activities than MB. MPMS always had the lowest efficiency in electron transfer ability using an aqueous or agarose containing reaction medium (SDH). With PVA in the reaction medium (SDH determination) PMS was clearly superior to MPMS. MB showed only a small transfer activity under these conditions because PVA seems to bind MB almost completely. It is concluded that in histochemistry an appropriate electron carrier and electron carrier concentration must be determined for different incubation conditions, tissues, tissue preparations and dehydrogenases studied. General statements about the efficiency or inefficiency of an electron carrier as a result of only one incubation condition does not seem to be justified.
D000818:Animals; D004396:Coloring Agents; D004579:Electron Transport; D006651:Histocytochemistry; D066298:In Vitro Techniques; D007668:Kidney / Q000201:enzymology*; D007700:Kinetics; D007770:L-Lactate Dehydrogenase / Q000378:metabolism*; D008297:Male; D008773:Methylphenazonium Methosulfate / Q000031:analogs & derivatives; D009206:Myocardium / Q000201:enzymology*; D009243:NAD / Q000378:metabolism; D010078:Oxazines; D010619:Phenazines; D051381:Rats; D011919:Rats, Inbred Strains; D013385:Succinate Dehydrogenase / Q000378:metabolism*; D013778:Tetrazolium Salts / Q000378:metabolism
D004396:Coloring Agents; D010078:Oxazines; D010619:Phenazines; D013778:Tetrazolium Salts; D009243:NAD; C033411:1-methoxyphenazine; D008773:Methylphenazonium Methosulfate; C005559:Meldola blue; D007770:L-Lactate Dehydrogenase; D013385:Succinate Dehydrogenase