Adult primary whole-brain cultures from Thy1-aSyn mice (Line 61) retain Parkinson's disease-relevant phenotypes

Primary cultures from fetal or postnatal rodent brain are widely used to study alpha-synuclein (aSyn)-associated mechanisms but poorly reflect age-dependent neurodegenerative processes. We established primary whole-brain cultures from adult Thy1-aSyn (Line 61) mice overexpressing human aSyn and compared them with postnatal and wild-type cultures. Cultures were maintained for 21 days and characterized by electrophysiology, immunohistochemistry, gene expression analysis, and protein biochemistry. Adult and postnatal cultures contained all major neural cell types, including neurons, microglia, astrocytes, and oligodendrocytes. ASyn overexpression was associated with reduced neuronal viability, altered neuronal firing, and increased microglial reactivity. Importantly, adult cultures retained the microgliosis observed in vivo in Thy1-aSyn mice. These findings establish adult Thy1-aSyn whole-brain cultures as a disease-relevant in vitro model for investigating aSyn-associated neuronal dysfunction, neuroinflammation, and cell-cell interactions and for evaluating therapeutic strategies.