BRAIN HEXOKINASE - IMMUNOHISTOCHEMICAL LOCALIZATION AT LIGHT MICROSCOPIC LEVEL

The indirect fluorescent antibody technique was used to localize hexokinase at the light microscopic level. Antibodies were produced in rabbits to soluble bovine brain hexokinase. Evidence of purity was obtained from agar gel diffusion and the precipitin test. The antiserum completely inhibited soluble hexokinase but only partially (60-70%) inhibited the activity associated with intact mitochondria or with the outer membrane fraction of mitochondria. When these antibodies were used to demonstrate the presence of hexokinase in thin sections of brain cortex, bright fluorescent granules were observed in the cytoplasm of nerve cells and in the intervening tissue. These corresponded in size and distribution to mitochondria demonstrated either by the succinate-dependent reduction of tetranitro blue tetrazolium or a classical supravital staining method. An unsuccessful attempt was made to confirm these observations in the electron microscope using the peroxidase-conjugated antibody technique. © 1969, American Chemical Society. All rights reserved.