Ultracytochemical Demonstration of Monoamine Oxidase Activity in Nervous and Non-nervous Tissue of the Rat

A method is described for the ultrastructural localization of the enzyme monoamine oxidase based on the formation of an osmiophilic formazan from the tetrazolium salt (2-(2'-benzothiazolyl)-5-styrl-3-(4'-phthalhydrazidyl) tetrazolium chloride) (BSPT) and using tryptamine as substrate. It was tested in rat heart, liver, adrenal cortex and medulla, kidney, pineal gland, cerebral cortex and cerebellar cortex and was found to provide good ultrastructural preservation, high contrast of the specific, enzyme-dependent precipitate and improved sensitivity. Monoamine oxidase activity was found in the mitochondrial outer compartment, nuclear envelope and endoplasmic reticulum in all organs studied; it was also seen occasionally in plasma membranes of liver and heart. Similar localizations were observed in interstitial cells and capillary endothelial cells. The enzyme-dependent nature of the observed precipitate was verified by control experiments consisting of incubation of overfixed tissue, omission of substrate from the incubation medium, and incubation in the presence of a specific, nonreversible inhibitor, pargyline. These studies confirm and extend the results of cell fractionation studies using liver, demonstrate the widespread distribution of monoamine oxidase in tissues, and suggest that monoamine oxidase plays important functional roles in extraneuronal cells.