Metabolite Profiling of Primary Neuronal and Glial Cell Cultures of the Mouse Cerebellum in an in vitro Ischemia Model

The relevance of research in the field of cerebral ischemia is driven by the need to identify therapeutic targets capable of preventing neuronal cell death, specifically through maintaining the balance of metabolic processes. Although significant shifts in the metabolite composition of brain tissues during ischemia have been previously detected, the analysis of metabolite changes in a heterogeneous tissue does not allow for the assessment of metabolic pathway activity in individual cell populations. This work was aimed to perform a non-targeted metabolomic analysis of primary neuronal and glial cultures of the mouse cerebellum in an in vitro ischemia model to identify specific metabolic pathways altered by oxygen-glucose deprivation (OGD) and reoxygenation. Neuronal and glial cultures were exposed to OGD followed by reoxygenation. Metabolite profiling was performed using UHPLC-MS with multivariate data processing algorithms to identify enriched metabolic pathways. Pronounced changes in metabolite profiles were shown in both neuronal and glial cultures in response to ischemic exposure. In both cell culture types, 15 potentially altered pathways from four classes of metabolic conversions were identified, including amino acid, glutathione, lipid, nucleotide, cofactor, and vitamin metabolism. These pathways may be involved in the regulation of energy metabolism, antioxidant defense, neurotransmitter balance, apoptosis, and neuroinflammation. The results obtained via non-targeted metabolomics methods allow for the identification of metabolic pathways of interest for subsequent targeted research in an in vitro model of nervous tissue ischemia.