TOLUENE INDUCED REMODELING OF CYTOSKELETON AND PROLIFERATIVE BEHAVIOR IN NEURAL STEM CELLS

Toluene is a widely abused inhalant whose impact on neural stem cells (NSC) and early neural lineage commitment remains poorly characterized. This study evaluated the concentration and time dependent effects of toluene on NSC proliferation, cytoskeletal organization, differentiation, and recovery capacity in vitro. NSC cultures were characterized by confocal immunofluorescence (MBP, GFAP, acetylated tubulin, DAPI), cell cycle analysis by flow cytometry, WST 8 viability/proliferation assays, and concentration–effect curves under increasing toluene exposure. Cytoskeletal integrity was examined by confocal microscopy (microfilaments, total and acetylated α tubulin, fascin), and protein expression was quantified by Western blot, including actin, GFAP, fascin, total and acetylated tubulin, and activation of p ERK1/2, p AKT and p GSK3β after 5 mM toluene at multiple time points. Additionally, we analyzed the impact of a toluene concentration curve on NSC differentiation into neuronal (NF), astroglial (GFAP) and oligodendroglial (MBP) phenotypes, as well as MBP positive process complexity, and implemented recovery paradigms (24–48 h) following acute toluene exposure. Overall, our findings indicate that toluene disrupts NSC proliferation and cytoskeletal architecture, alters lineage specific marker expression and process complexity, and that these alterations show only partial normalization after recovery intervals, suggesting persistent vulnerability of the NSC compartment to inhalant exposure.