Temperature-dependent regulation of PLP/DM20 and CNP gene expression in two conditionally-immortalized jimpy oligodendrocyte cell lines.

Abstract

We conditionally immortalized jimpy primary oligodendrocytes (ODCs) with the temperature-sensitive SV40 large T antigen. Two cell lines (clones JP1.1 and JP1.2) were generated that expressed a number of ODC markers. Both jimpy cell lines expressed DM20 mRNAs at the proliferative temperature of 34 degrees C, but not at the "differentiation" temperature of 39 degrees C. Interestingly, at 39 degrees C neither cell line appeared to differentiate further, and neither survived longer than 7 days, in contrast to other ODC cell lines from normal animals that survive many weeks at 39 degrees C. These findings are not consistent with the notion that a PLP/DM20 gene product is the cause of oligodendrocyte cell death in jimpy, since neither jimpy cell line survived at 39 degrees C, and neither line expressed PLP or DM20 proteins. Analysis of the expression of the CNP (2'3' cyclic nucleotide-3'-phosphodiesterase) gene indicated that in both cell lines only one of the two CNP isoforms was expressed at 34 degrees C. Raising the temperature to 39 degrees C caused a greater reduction in the levels of CNP protein than CNP mRNA. Taken together, the DM20 and CNP data suggest that at least some of the decline in myelin/oligodendrocyte components observed in jimpy brains may not be due simply to fewer mature oligodendrocytes, but also to a down regulation of expression of these genes at several levels including transcriptional and post-transcriptional events. Our results provide two cell models for in vitro investigations into the nature of the jimpy mutation at several cellular and molecular levels.