The formation of cerebrosides by the galactosylation of ceramides was investigated in a microsomal fraction prepared from sciatic nerves of normal and Trembler mice. In the control, cerebroside synthesis is observed in the presence of uridine-diphosphate-galactose both from endogenously synthesized [1- 1 4 C]stearoyl-sphingosine (C18-ceramide), and from [1- 1 4 C]lignoceroyl-sphingosine (C24-ceramide). Cerebroside formation is also demonstrated by studying the galactosylation of exogenous ceramides with UDP[1- 1 4 C]-galactose. In the mutant, only trace amounts of labeled cerebrosides are formed from labeled stearoyl-sphingosine, whereas with lignoceroyl-sphingosine, no cerebroside synthesis is detected under conditions allowing their formation in the control. However, a higher rate of synthesis of short acyl chain-glucosyl ceramides is observed in the Trembler samples.