Abstract
Early‐life exposure to general anesthetics has been linked to long‐term neurodevelopmental risks, yet the underlying mechanisms remain unclear. Using a mouse model, we show that neonatal isoflurane exposure at Postnatal Day 7 leads to age‐dependent behavioral alterations, changes in cortical neuronal abundance and layer‐specific neuronal density, and dysregulation of cytoskeleton‐associated proteins. Isoflurane‐exposed mice displayed increased exploratory behavior during early adolescence but developed marked recognition memory deficits by late adolescence. Histological analyses revealed layer‐specific disruption of neuronal distribution in the sensory cortex, characterized by reduced neuronal density in layers II–IV and age‐dependent changes in inhibitory interneuron composition. Mechanistically, isoflurane reduced the expression of the microtubule plus‐end tracking protein EB3 and the transcription factor FOXG1. FOXG1 co‐localized with EB3 and β‐tubulin in the cytoplasm, supporting a noncanonical, cytoskeleton‐associated role. Together, these findings indicated that early isoflurane exposure is associated with altered neuronal density and dysregulation of the cytoskeleton‐associated proteins FOXG1 and EB3, suggesting a potential link between anesthesia exposure and long‐term neurodevelopmental alterations.