The role of glia in amyotrophic lateral sclerosis (ALS) is undeniable. Their disease-related activity has been extensively studied in the spinal cord, but only partly in the brain. We present herein a comprehensive study of glia in the cortex of SOD1(G93A) mice-a widely used model of ALS. Using single-cell RNA sequencing (scRNA-seq) and immunohistochemistry, we inspected astrocytes, microglia, and oligodendrocytes, in four stages of the disease, respecting the factor of sex. We report minimal changes of glia throughout the disease progression and regardless of sex. Pseudobulk and single-cell analyses revealed subtle disease-related transcriptional alterations at the end-stage in microglia and oligodendrocytes, which were supported by immunohistochemistry. Therefore, our data support the hypothesis that the SOD1(G93A) mouse cortex does not recapitulate the disease in patients, and we recommend the use of a different model for future studies of the cortical ALS pathology.

2nd Faculty of Medicine Charles University 5 Uvalu 84 15006 Prague Czech Republic

Department of Cellular Neurophysiology Institute of Experimental Medicine Czech Academy of Sciences Videnska 1083 14220 Prague Czech Republic

Department of Functional Organization of Biomembranes Institute of Experimental Medicine Czech Academy of Sciences Videnska 1083 14220 Prague Czech Republic

Department of Informatics and Chemistry Faculty of Chemical Technology University of Chemistry and Technology Technicka 5 16628 Prague Czech Republic

Faculty of Science Charles University Albertov 6 12800 Prague Czech Republic

Laboratory of Gene Expression Institute of Biotechnology CAS BIOCEV Prumyslova 595 25250 Vestec Czech Republic

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ALS-like pathology diminishes swelling of spinal astrocytes in the SOD1 animal model

Reactive gliosis in traumatic brain injury: a comprehensive review