The plasma membrane is characterized by an asymmetric distribution of phospholipids, maintained by three enzyme classes: flippases, floppases, and scramblases. Phosphatidylserine (PS) is confined to the inner leaflet of the plasma membrane under normal conditions, and its externalization has been linked to both apoptotic and non-apoptotic processes within the central nervous system (CNS). Further, neuronal PS exposure can be transient in stressed but viable cells. Thus, interference with PS-triggered phagocytosis of neurons has been suggested to be beneficial in CNS disorders. The mechanism of action, expression levels, and response to stimuli of PS-shuttling enzymes in neurons remain poorly characterized.Using different imaging approaches, we found that an excitotoxic insult to primary hippocampal neurons triggers PS exposure. The exposure could be identified in the form of hotspots, primarily localized near spine protrusions and/or dendritic branching points. Notably, this phenomenon was accompanied by a reduction in expression levels of Atp8a2; a flippase responsible for the inward translocation of PS.We demonstrated that silencing Atp8a2-activity results in an increased PS exposure, particularly at dendritic branching points, coupled with reduced dendritic complexity and neurite length. Moreover, diminished Atp8a2 activity intensifies neuronal vulnerability to toxic insults both in-vitro and in-vivo, while boosting Atp8a2 expression provided neuroprotection.As mutations in Atp8a2 in humans have been linked to mental retardation, motor deficits and optic atrophy in humans, our work provides mechanistic insights into these genetic disorders.