Expression of galactocerebroside in developing normal and jimpy oligodendrocytes in situ

Abstract

Intense and specific immunostaining of oligodendrocytes in vivo has been obtained for the first time using antibodies to galactocerebroside. We have examined the differentiation of oligodendrocytes in normal mice and then compared their differentiation to the myelin-deficient mouse jimpy, using immunoperoxidase, immunogold and immunofluorescence labelling techniques. We also compared staining for galactocerebroside with staining obtained using antibodies to myelin basic protein, carbonic anhydrase II, 2', 3'-cyclic nucleotide 3'-phosphohydrolase and proteolipid protein. The results of this comparative study confirm previous tissue culture studies and show that galactocerebroside is specific for oligodendrocytes in situ. As in tissue culture, galactocerebroside is one of the earliest oligodendrocyte markers to be expressed, making it an important marker for studying the differentiation of this cell type. The shape of oligodendrocytes in situ changes distinctly with time, shifting from an early stellate form with numerous spidery processes to a cell with a few processes radiating from the perikaryon. These morphological changes are observed for both normal and jimpy mice and they parallel those described in vitro. Oligodendrocytes in jimpy mice express most myelin markers, but the staining within the cells is generally less intense than in normal oligodendrocytes and the antigens are restricted to the cell body and processes without being incorporated into myelin sheaths. Quantification of the number of oligodendrocytes stained for galactocerebroside in normal and jimpy mice show that their number is not reduced in the corpus callosum and cerebellum during the first 2 weeks postnatal. This finding shows that many cells in jimpy mice which were considered to be unclassifiable by the application of morphological criteria have, in fact, differentiated to the stage where they are galactocerebroside-positive.