Localization of cholinesterases in adult mammalian brain cell cultures

Abstract

1. AChE was localized in all neuronal elements and BuChE in glial elements in the explants and migratory zones of whole mount adult rabbit and human cortex cultures maintained up to 6 months. Koelle's technique ( Koelle, 1955) with AChTh and BuTh as substrates and suitable inhibitors was used. Eserine (10 −5 to 10 −3 M) inhibited cholinesterases. DFP (10 −8 to 10 −6 M) and RO2-0682 (6 × 10 −8M) inhibited BuChE. 2. AChE was present in the perikarya and fibers and was found to highly concentrated in axonal endings, especially presynaptic endings. 3. Cultures from human subcortical white matter after 2 months in culture showed an absence of AChE but the presence of BuChE in glial elements. AChE and BuChE could be localized in corresponding gray matter cultures. 4. Sodium barbital decreased the intensity of staining for AChE and increased the staining of glial elements for BuChE. Epinephrine reduced the stain intensity of AChE in the perikarya of neurons while at the same time the fibers and boutons stained intensely.