[Immunohistochemical study of ischemic neuronal damage with antiserum to tubulin, a microtubular protein].
The limitation of conventional histological methods to demonstrate ischemic change of neurons in early phase has been a major drawback in histopathological and pathophysiological studies of cerebral ischemia. Cellular metabolism is disturbed immediately after cessation of the regional circulation and rapid alterations in macromolecular and ultrastructural integrity in neurons may take place before any evidence of histopathological changes could be detectable. To demonstrate ischemic change of neurons more sensitively on a histological level, we applied immunohistochemical method using antiserum to tubulin, a protein of microtubules. As this organelle has been implicated in several important cellular functions such as control of cell shape, intracytoplasmic transport of materials or synaptic transduction, immunohistochemical alterations in microtubules may indicate structural as well as functional damage of neurons. In order to study the ischemic change in neurons, the posterior communicating artery of a gerbil brain was occluded by the method previously reported by us, and the hippocampus, which is one of the most vulnerable structures of the brain to ischemia, was observed. Five or 30 minutes after occlusion, animals were sacrificed by decapitation. Brains were removed, cut coronary vessels and fixed in ethanol-acetic acid (95:5). Tubulin used for this study was extracted from normal gerbil brains and specific antiserum was raised in goats Peroxidase-antiperoxidase method was performed on paraffin sections. Immunohistochemical distribution of tubulin in a normal gerbil brain demonstrated by the present method was in good accordance with the reported electronmicroscopical distribution of microtubules.(ABSTRACT TRUNCATED AT 250 WORDS)