A marker for oligodendrocytes and its relation to myelinogenesis: An immunocytochemical study with experimental allergic encephalomyelitis serum and C.N.S. cultures

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Abstract
To investigate a possible marker for oligodendrocytes and its relation to myelinogenesis, experimental allergic encephalomyelitis (EAE) serum has been used to study C.N.S. cultures from the time of explantation to maturity at 26 daysin vitro (DIV). Cultures of foetal mouse spinal cord were exposed for 1 h to heated (complement-inactivated), rabbit anti-bovine white matter (WM-EAE) or control serum, fixed and processed by an immunoperoxidase technique for demonstrating bound immunoglobulin (Ig) by light and electron microscopy. From 5 to 26 DIV, cells morphologically identical to oligodendrocytes displayed binding of Ig to the plasmalemma of the cell body and its processes. At 5 DIV, immunoreactive oligodendrocytes had a large nucleus and nucleolus, prominent Golgi apparatus, and microtubules but no filaments. Occasionally a centriole was present, suggesting an early stage of differentiation. In myelinated cultures (from 11–12 DIV onwards), reaction product was present on the oligodendroglial outer plasmalemma apposed to myelin and along the outer loop. Sometimes it extended into the external mesaxon, outer layer of myelin, inner mesaxon and periaxonal space. No other structures were reactive, and oligodendroglia did not bind control Ig. These findings indicate that WM-EAE serum can be used as a marker for oligodendrocytes in cultures from 5 DIV onwards. The findings that oligodendrocytes acquire the antigen(s) prior to myelination and that the antigen(s) is localized on the plasmalemma of the inner and outer loops of actively myelinating oligodendroglial processes suggest that the antigen(s) may have a role in oligodendrocyte maturation and myelinogenesis. The antigen(s) involved is not yet established, but it is probably not myelin basic protein. This marker should prove useful in studies of C.N.S. development and the demyelinating diseases.

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