Ultrastructural immunocytochemistry of γ-aminobutyrate in the cerebral and cerebellar cortex of the rat

Abstract

γ-Aminobutyrate containing structures in the cerebral and cerebellar cortex of the rat were visualized by an immunocytochemical method using glutaraldehyde fixation and an antiserum developed against a γ-aminobutyrate-glutaraldehyde-protein conjugate. Labelled elements (perikarya and cell processes) were observed to be distributed throughout the layers of the cerebral cortex in a pattern similar to that described using glutamate decarboxylase immunocytochemistry. The morphological features of many immunoreactive cell bodies were typical of stellate neurons. In the cerebellar cortex, Purkinje, basket, Golgi and stellate, cell bodies were found to be immunoreactive along with numerous labelled neuronal processes. At the ultrastructural level, the labelled processes in both areas corresponded to immunoreactive dendrites and fibres. Labelled synaptic boutons, generally of the symmetrical type, could also be seen in contact with positive or negative cell bodies and dendrites. In the cerebellum, glomeruli could be clearly identified including mossy fibres surrounded by unlabelled dendrites in contact with immunoreactive terminals. At the subcellular level in both brain regions, the areas occupied by the Golgi apparatus were never labelled, although the nuclei had varied reactions. The strong glutaraldehyde fixation that limits the diffusion of γ-aminobutyrate limits also antibody diffusion. However, this fixation is compatible with a good morphological preservation and should enable immunocytochemistry studies to be compared to other methods such as autoradiography.