Light and electron microscopic localization of a cell surface antigen (NG2) in the rat cerebellum: association with smooth protoplasmic astrocytes

Abstract

Immunofluorescence and immunoperoxidase techniques were used to localize a cell surface chondroitin-sulfate proteoglycan antigen, termed NG2, in the developing and adult rat cerebellum. In the adult, both polyclonal and monoclonal anti-NG2 antibodies labeled cells throughout the cerebellar cortex, with the labeled cells being especially prominent in the molecular layer. The labeled cells had small, irregularly shaped cell bodies from which thin highly branched processes radiated in a stellate array. The NG2-labeled cells were not labeled with antibodies against glial fibrillary acidic protein (GFAP), vimentin, or S-100 protein, intracellular markers for astrocytes. However, electron microscopic immunocytochemical analysis of NG2 immunoreactive cells revealed a cell morphology consistent with that of protoplasmic astrocytes. Labeled cell bodies contained a thin rim of organelle-poor cytoplasm surrounding a euchromatic nucleus. Thick processes originating from the cell soma tapered to form thin branches with highly irregular surface contours that extended between adjacent neuronal elements. The labeled processes did not form synapses in the neuropil, and no synaptic profiles onto anti-NG2-labeled cell bodies or processes were observed. Thus, we conclude that the NG2 antigen is a cell surface marker for a class of smooth protoplasmic astrocytes. Immunoreactive cells were seen in the developing cerebellum beginning at embryonic day 16. The number of labeled cells increased during the early stages of cerebellar development, reaching a peak at about postnatal day (PND) 4 or 5 and declining thereafter. In the developing cerebellum, labeled cells lying within the forming molecular layer resembled the cells seen in the adult, whereas cells lying deeper within the folia had an immature appearance with fewer processes and less branching. This apparent gradient of morphological maturation suggests that an interaction with parallel fibers in the developing molecular layer may play a role in the terminal cytodifferentiation of the NG2-labeled smooth protoplasmic astrocytes.