Synthesis of Gluco‐ and Galactocerebrosides in Bovine Neurones and Oligodendroglia

Oligodendroglial and neuronal perikarya have been isolated from fresh and frozen bovine brain and used to investigate the synthesis of cerebrosides from UDP-hexoses and ceramides. The radioactive cerebrosides produced have been identified by TLC of the intact lipids on borate-silica gel and by a combination of acid hydrolysis and ion-exchange chromatography of the liberated hexoses. Incubation of either neuronal or oligodendroglial homogenates with UDP-galactose (UDP-gal) and mixed ceramides (hydroxy plus nonhydroxy fatty acids) leads to the synthesis of hydroxy fatty acid galactocerebroside. This lipid is also formed by both neuronal and oligodendroglial homogenates if the UDP-gal is replaced by UDP-glucose (UDP-glu). Formation of glucocerebroside has been observed only in the presence of neuronal homogenates and UDP-glu. Electron microscopic examination of the isolated cell preparations has shown that, although the oligodendroglia are relatively pure, the neurones are contaminated with oligodendroglia, which may account for some of the hydroxy fatty acid galactocerebroside synthesis. When unlabelled UDP-gal is included in the incubations containing cell homogenates, mixed ceramides, and labelled UDP-glu, galactocerebroside formation is greatly reduced, but there is comparatively little effect on glucocerebroside synthesis. In addition, experiments using D-[14C]galactose 1-phosphate and UDP D-[6-3H]glucose simultaneously have shown that the rate of formation of tritiated cerebroside is much greater than that of the 14C-labelled compound.(ABSTRACT TRUNCATED AT 250 WORDS)