A new subdivision of the periaqueductal gray (PAG) based on its myelin content is described. Using a methacrylate-resin embedding technique for high resolution light microscopy, a myeloarchitectonic map of the normal rat PAG was made. Six main columns were distinguished. Column I is the central thin subependymal layer, in which myelinated axons are virtually absent. This layer is surrounded by column II with many rostrocaudally oriented fibers. Lateral and dorsolateral to column II lies column III, which contains rostrocaudally as well as tangentially oriented fibers. Column III is divided into a ventral and a dorsal part by column IV This column is located in the dorsolaferal part of the PAG and contains very few, mostly tangentially oriented fibers. Column V is located ventrally and contains a mixture of rostrocaudally and tangentially oriented fibers. Column VI is only present in the most caudal part of the PAG and contains mainly tangential fibers.