Abstract
Oligodendrocytes were isolated from the white matter of ox brains. Light microscopy revealed that the cells were greater than or equal to 90% phase-bright with a mean diameter of 7.6 micron. Transmission electron microscopy was employed to identify the classic morphology associated with mature oligodendrocytes. Homogenates of the isolated cells showed negligible activity of neuronal and astrocytic cell markers. Using a suspension culture method cells were incubated with [14C]glucose. This simple precursor labelled the five complex lipids choline glycerophospholipid, ethanolamine glycerophospholipid, inositol glycerophospholipid + serine glycerophospholipid, and the two cerebroside species. The incorporation of label was shown to be dependent on glucose concentration, protein concentration, and the length of incubation. In addition the glucose uptake blocker phloretin (1 mM) reduced the degree of labelling by up to 97%, and the metabolic poisons KCN (1 mM) and iodoacetate (0.5 mM) had varying deleterious effects on the amounts of labelling of the five lipids measured.
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