Endogenous peroxidase in mast cells localized with a semipermeable membrane technique

Abstract

Hamster mast cells have been found to give strong peroxidatic reactions at pH 5, 7.5 and 10 when sections of skeletal muscle are incubated for 2.5 h in the dark at room temperature on semipermeable membranes covering a gelled incubation medium consisting of 0.01% hydrogen peroxide, 5.5mM diaminobenzidine and 1.36% agar dissolved in Universal buffer. The technique is very efficient: with it, all mast cells react in marked contrast to the negative reaction they usually give with conventional techniques. The peroxidatic reactions are abolished if tissues are perfused beforehand with either aminotriazole or KCN but not if these inhibitors are incorporated in the gelled incubation medium. This and other evidence suggests that the mast cell reactions are not due to either catalase or haemoglobin adsorbed onto mast cell granules from lysed red blood cells. Skeletal muscle fibres do not exhibit any visible peroxidase activity with the membrane technique.