TRANSFECTION OF TRANSFORMED SHIVERER MOUSE GLIAL-CELL LINES

In the course of studies on glial cell differentiation in the mouse mutant shiverer, we have established by infection with a temperature-sensitive retrovirus encoding the SV40 T antigen a few glial cell lines that divide rapidly at 33°C, the permissive temperature, and whose phenotypes at the nonpermissive temperature (39°C) resemble either astrocytes or oligodendrocytes. One astrocyte-like clone (clone SF11) expresses glial fibrillary acidic protein at 33°C and expresses this marker strongly at 39°C as well. One oligodendrocyte-like clone (DM5) that we have succeeded in maintaining at 33°C can be induced to express galactocerebroside and 2',3'-cyclic nucleotide 3'-phosphodiesterase at the elevated temperature but never expresses the more differentiated markers of the myelination state, such as the myelin proteolipid protein or the myelin-associated glycoprotein. Clone DM5, as is the case with other oligodendrocyte clones we have prepared by the method described here, is quite fragile and survives for only several (5-6) days at the higher temperature. Both clonal lines can serve as host cells for expressible cDNAs introduced by transfection, such as the neurofilament protein, NF-M, and the small myelin basic protein.