A direct comparison of GFAP immunocytochemistry and GFAP concentration in various regions of ethanol-fixed rat and mouse brain.

Glial fibrillary acidic protein (GFAP) immunostaining is the most commonly used method to examine the distribution of astrocytes and the hypertrophy of astrocytes in response to neural degeneration or injury. More recently, a variety of biochemical assays for GFAP have been developed. Both qualitative immunocytochemical evaluations of GFAP and quantitative biochemical measurements of GFAP have been used to examine the regional distribution of GFAP within the central nervous system (CNS). The former method has largely been based on aldehyde-fixed tissue, while the latter approach has been based on the use of fresh tissue extracts or homogenates. In the present study, we used ethanol as a fixative to permit both immunocytochemical and biochemical procedure to be carried out on brain tissue from a single animal. Normal adult rats and mice were perfused with a 70% ethanol/saline solution, and each brain was hemisectioned. The concentration of GFAP was measured in regions of 1 hemisection, using an enzyme-linked immunosorbent assay (ELISA), while the other hemisection was used for GFAP immunostaining. Regional differences occurred in the brains of both species, with the highest concentration of GFAP found in the brainstem, and the lowest concentrations found in the striatum and cortex. The specific patterns of GFAP immunoreactivity corresponded to regional concentrations in most brain area of both species. These data show that it is possible to assay GFAP concentrations in tissue prepared for immunocytochemical analysis, providing both qualitative and quantitative information from one set of tissue.