Quantitative chemical analysis of demyelination in cultured mouse spinal cord

These studies were designed to determine whether chemical parameters can be used to quantify the demyelinative potency of anti-CNS sera or other demyelinating factors as revealed by cultured nerve tissue. The activity of 2',3'-cyclic nucleotide 3'-phosphohydrolase (CNPase) and the amount of some major myelin lipids were measured in cultured embryonic mouse spinal cord during different stages of demyelination. Cultures exposed to anti-white matter serum for various times (6-60 h) or various concentrations (2.5-25%) showed a decrease (up to -70%) in CNPase activity. This change paralleled the degree of demyelination as evaluated by light microscopic examination. Cultures exposed to normal rabbit serum under the same conditions did not show changes in the amount of myelin or in the activity of CNPase. Conversely, changes in the lipid content did not correlate with the morphologic evaluation of demyelination. Glycolipids were found to decrease in cultures exposed to anti-white matter as well as in those exposed to normal rabbit sera. The decrease in cerebrosides was higher with anti-white matter serum than with normal serum, but sulfatides were altered in similar amounts in both groups. In contrast, under the same conditions, the amount of total protein or other lipid such as total cholesterol is not altered. Therefore, biochemical analysis of some myelin components could constitute a more sensitive and reliable quantitative method for early evaluation of the time course of demyelination in cultured nerve tissues.