Regional patterns of neurodegeneration in a mouse model of proteinopathy

Abstract

The aggregation of misfolded proteins is a hallmark of many neurodegenerative diseases, suggesting shared pathological mechanisms. However, the pathways by which protein misfolding in these proteinopathies lead to neuronal death remain unclear. Proteinopathies can be modelled in transgenic animals by expressing disease-causing mutations that promote protein aggregation, or in wild-type animals by injecting misfolded proteins (e.g. RML scrapie) that spread in a prion-like manner and recapitulate key neurodegenerative features, including gliosis, ER stress, and neuronal loss. Here, we map region-specific histopathological features of scrapie-induced neurodegeneration in the hippocampus, thalamus, cortex, and cerebellum during early (12 weeks post-inoculation) and late (20 weeks) stages of disease. Using a streamlined time-efficient protocol, we achieve reproducible paired sample collection and high-quality immunohistochemistry that is compatible with best practice in decontamination and containment. We found that among the tested markers of early pathology, thalamic astrocytic activation and spongiform degeneration were the most sensitive. By the late stage, there was widespread upregulation of IBA1+ microglia and GFAP+ astrocytes, accompanied by strong immunoreactivity of lysosomal marker LAMP1. LAMP1 expression in healthy brains was largely neuronal, but by 20 weeks it was significantly upregulated in astrocytes, suggesting their involvement in lysosomal pathology. The ER stress marker p-PERK was elevated in CA1/CA3 pyramidal neurons but minimal in the thalamus and cerebellum, where neuronal loss was most pronounced, suggesting region-specific mechanisms of degeneration. Overall, the thalamus and hippocampal CA1/CA3 areas exhibited the greatest pathological burden. Our shorter time-course, new pathological insights and safe handling protocols, and improved welfare, supports broader adoption of the RML scrapie model for resource-efficient studies of neurodegeneration and its prevention.