In the course of scrapie, a transmissible spongiform encephalopathy caused by an unconventional agent, a normal cellular protein is converted to an abnormal form that copurifies with infectivity and aggregates to form deposits of amyloid. We have used immunocytochemistry and methods that enhance detection of amyloidogenic proteins to investigate the types of cells in the central nervous system which are involved in the formation of the abnormal scrapie-associated protein. We show that this protein accumulates in astrocytes prior to the cardinal neuropathological changes in scrapie--astrogliosis, vacuolation, neuron loss, and amyloid deposition. These findings implicate the astrocyte in the formation of the scrapie isoform of the prion protein and amyloid in scrapie and suggest that this cell type might also be involved in the replication of the scrapie agent.