Developmental expression of neutral monoglycosylceramides (MGCs) has been examined in rat brain from embryonic day 15 (E15) to postnatal day 30 (P30) and adulthood. To this point, glucosylceramide (GlcCer) is the only MGC that has been characterized in embryonic brain. Galactosylceramide (GalCer) appears at P1, increases with age until P25 and remains constant thereafter. The developmental occurrence of GlcCer and GalCer agrees well with their respective glucosyl- and galactosyltransferase activities. Cerebroside fatty-acid and base compositions, examined by gas chromatography, also change during development. Several alkali-labile fast-migrating cerebrosides (FMCs) with a higher thin-layer chromatography RF than GalCer/ GlcCer are expressed early at P10, increase in concentration with age (P25-P30) and are unchanged until maturity. They are derivatives of GalCer. By employing a newly developed neutral methylation procedure, we have confirmed the structure of one of the FMCs as 6-acylGalCer. A reduction in brain FMC concentrations along with GalCer in murine genetic dysmyelinating disorders (jimpy and quaking) further supports the conclusion that they are myelin constituents.