The synaptic localization of alpha-internexin, a brain-specific intermediate filament protein, was investigated immunohistochemically in the rat brain. The specificity of the antibody used in this study was confirmed by Western blotting and the antibody specifically reacted with alpha-internexin in the neurofilament preparation and in the postsynaptic density (PSD) fraction. The alpha-internexin immunoreactivity was distributed in neurons, especially in the somata and dendrites, throughout the cerebral cortex. Immunoelectron microscopic examination showed the immunoreactivity in the PSD, while neurofilament M was not in the PSD. Thus alpha-internexin and neurofilament M are differentially localized in neuronal cells. Alpha-internexin content in the PSD fraction was relatively high even before the period of synaptogenesis and the content in the fraction was unchanged between young and adult rats (2-6 weeks old). These results suggest a role of alpha-internexin for early development and organization of the PSD.