Abstract

Introduction. Vitamin C is an antioxidant molecule found in high concentrations in the central nervous system. Under physiological conditions predominates the reduced form, ascorbic acid (AA), which is incorporated by neurons through co-transporter sodium ascorbate 2 (SVCT2). When Vitamin C fulfills his antioxidant function, dehydroascorbic acid (DHA) is produced, which diffuses into nerve cells by GLUTs transporters. Under conditions of oxidative stress DHA concentration increases accumulates in the neuron with unknown effects. The aim of this study was analyze neurons under oxidative stress when were preloaded with AA in the presence/absence of vitamin C recycling cells, astrocytes. Materials and Methods. Neuro2a cell line (N2a) and rat cortical neurons were supplemented with AA and thereafter treated with H2O2. Cell viability was quantified by XTT method. Co-culture experiments were performed in presence/absence of astrocytes. In addition, cell morphology and distribution of Vitamin C transporters was analyzed by spectral microscopy, super-resolution SIM and live cell. Results. Oxidation of vitamin C generates neuronal death, which is prevented by astrocytes cytochalasin B dependently. Before neuronal death, the cells modifies the distribution of GLUT1/SVCT2. Furthermore, we note decrease of mitochondrial activity (MitoTracker, red) with live cell experiments, condition finished in permeabilization of the plasmatic membrane and the incorporation of phalloidin (green). Discussion. DHA intracellular generation in absence of astrocytes produces neuronal death. This process modifies neuronal cell morphology and distribution SVCT2/GLUT1 oppositely in the cytoplasm of neuron. FONDECYT 1140477, CMA BIO BIO, CONICYT ECM-12.