ELECTRON MICROSCOPE STUDIES OF GLUTAMIC OXALACETIC TRANSAMINASE IN RAT LIVER CELL

Abstract

Liver tissue of the rat, fixed in glutaraldehyde and formaldehyde, was incubated in a medium which consisted of 20 mM L-aspartic acid, 2 mM α-ketoglutaric acid, 50 mM imidazole and 6 mM lead nitrate at pH 7.2–7.4. The electron-opaque precipitates, due to glutamic oxalacetic transaminase activity in liver cells, were found to be localized to the cristae and surface membranes of the mitochondria, the limiting membrane of the microbodies, and the nuclear membrane. Sucrose storage and trauma resulted in altered morphology and diminished final product intensity in mitochondria, but the microbody enzyme activity disappeared completely under these conditions. These distinctive responses of enzymatic activity are considered to indicate a difference in either the enzyme protein or its membrane attachment to these two sites. The use of a buffered dehydrating ethanol series to prepare tissue blocks for electron microscopy appeared to result in more precise intracellular localization of enzymatic reaction product.