The role of mouse strain and the influence of gender on demyelination were explored for the first time in SJL mice using the cuprizone intoxication model. We document here that SJL mice display a unique pattern of demyelination that did not follow the profile that is well-characterized in C57BL/6 mice. The SJL mice did not readily demyelinate at the midline within the corpus callosum but showed greater demyelination immediately lateral to midline. During continuous exposure to cuprizone, demyelination was not complete and appeared to plateau after week 7. Importantly, female mice were partially resistant to demyelination, whereas male mice were more severely demyelinated. Differences in the number of mature oligodendrocytes were consistent with the extent of demyelination; however, microglia, astrocyte and oligodendrocyte precursor cell populations did not differ between male and female mice. Thus, genetic factors and gender influence susceptibility to demyelinating disease in the cuprizone model, which may provide additional insights into the variability observed in human demyelinating diseases such as multiple sclerosis.