EFFECTS OF COMPOUND 48/80, CHYMOTRYPSIN AND ANTISERUM ON ISOLATED MAST CELLS UNDER AEROBIC AND ANAEROBIC CONDITIONS

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Abstract
Degranulating actions of compound p-methoxyphenethylmethylamine condensation product with formaldehyde (48/80), [alpha]-chymotrypsin and anti-serum were studied on mast cells isolated from rat peritoneal fluid under aerobic and anaerobic conditions. Anaerobic condition has been achieved by bubbling of nitrogen gas, before introduction of mast cells, through the incubation medium in Carrel''s flasks containing a small piece of yellow phosphorus. The presence of a small amount of bovine serum albumin in the medium has been confirmed to be effective for preserving the responsiveness of mast cells. There are some differences between these degranulating agents in the morphology of the affected mast cells and lag phase of the reaction. Degranulating actions of these agents are all dependent on temperature and pH. Ca2+ are essential for the actions of [alpha]-chymotrypsin and anti-serum on isolated mast cells but not for compound 48/80, while for the actions on mesentery mast cells all the agents require Ca2+. Ca2+ are also involved in complement activation in the action of anti-serum. Under aerobic condition the actions of these three degranulating agents are not influenced by the presence or absence of glucose in the medium. In anaerobiosis the actions of compound 48/80 and [alpha] -chymotrypsin are completely inhibited when the medium does not contain glucose, although this is not for the action of anti-serum owing to glucose contained in itself. KCN and 2,4-dinitro-phenol completely inhibit the actions of these three degranulating agents in the glucose-free medium in air phase. All these inhibitions are largely counteracted by the presence of a small amount of glucose in the medium, and this "glucose effect" is markedly blocked by phlorizin and 2-deoxyglucose, while the effect of ouabain is uncertain as far as isolated mast cells are concerned. Iodoacetate inhibits the degranula-tion irrespective of presence or absence of glucose. Mg2+ have no effect on the anoxic inhibition of degranulation. These results indicate that the degranulations of mast cells in response to these three agents all rely on energy generation, which is a function localized in mast cells, and that in the presence of exogenous glucose glycolysis takes part in supplying energy required for anoxic degranulation, alternating the aerobic oxidation of endogenous substrates.