Modulation of autophagy in a mouse model of cuprizone-induced demyelination

Multiple sclerosis (MS) is a chronic immune-mediated disorder affecting the central nervous system (CNS), characterized by inflammation, demyelination, and neuronal loss. Although inflammatory activity drives lesion formation, MS is now recognized as a disease involving both inflammatory and neurodegenerative mechanisms (Haki et al., 2024). Optic neuritis is the first clinical manifestation of MS in about 40% of patients and it occurs in up to 70% over the course of the disease (Tong et al., 2025). Autophagy is a key homeostatic process involved in degradation and recycling of cellular components, essential for neuronal integrity (Li et al., 2022). Alterations in this process have been associated with several neuroinflammatory and neurodegenerative conditions; however, its contribution to MS pathology remains unclear (Misrielal et al., 2020). The present study aims to evaluate the regulation of autophagy in the optic nerve and corpus callosum in a cuprizone-induced mouse model of MS. Demyelination was induced by feeding 8-week-old male C57BL/6J mice a diet containing 0.2% (w/w) cuprizone for 8 weeks. Protein expression levels were analyzed by western blotting and immunofluorescence. Proteomic profiling was performed using mass spectrometry. Cuprizone treatment induced a marked reduction in myelin basic protein (MBP) in both the corpus callosum and optic nerve, together with increased glial fibrillary acidic protein (GFAP). In the corpus callosum, levels of autophagy markers (i.e. p62 and LC3-I/II) were increase. PINK1 and Parkin expression was significantly upregulated, while optineurin remained unchanged. In the optic nerve, Parkin was increased, with no changes observed in retinal tissue. Proteomic analysis of the corpus callosum identified 5,987 proteins, of which 139 were significantly modulated by cuprizone treatment. These preliminary findings indicate dysregulation of autophagy and mitophagy in the corpus callosum and optic nerve during cuprizone-induced demyelination and support further investigation of these pathways as modulators of neurodegeneration in MS.