Titration of cuprizone induces reliable demyelination

Abstract

Multiple Sclerosis (MS) is a chronic inflammatory disease of the central nervous system. Cuprizone-induced demyelination, wherein mice are fed a diet containing the copper chelator cuprizone, is a well-established model that replicates key features of demyelination and remyelination. However, the dose–response relationship of cuprizone is complex; high concentrations can induce toxicity, whereas low doses may fail to produce reliable demyelination across subjects. This study aimed to investigate whether titration of the cuprizone concentration results in reliable acute demyelination and weight stabilization. To this end, experimental animals were intoxicated with cuprizone over a period of 5 weeks to induce acute demyelination. In one group, during the first 10 days, the initial cuprizone dose was gradually reduced until the experimental animals showed stable weights. Another group was subjected to a continuous cuprizone intoxication protocol without adaptions. Histological analyses were performed to quantify the extent of demyelination and glia activation. Animals of both groups experienced significant weight loss. Histological analyses revealed, despite adopting the cuprizone concentration, substantial demyelination of various brain regions, including the corpus callosum. This pattern was consistent across multiple staining methods, including anti-proteolipid protein (PLP), anti-myelin basic protein (MBP), and luxol-fast-blue (LFB) stains. Additionally, grey matter regions, specifically the neocortex, demonstrated significant demyelination. Accompanying these changes, there was notable activation and accumulation of microglia and astrocytes in white and grey matter regions. These histopathological changes were comparably pronounced in both cuprizone-treated groups. In summary, we demonstrate that titration of cuprizone is a reliable approach to induce acute demyelination in the mouse forebrain. This work represents a significant step toward refining animal models of MS, contributing to the broader effort of understanding and treating this complex disease.