Isoflurane-induced spine loss is reversible.

<p>Hippocampal neuron cultures transfected with eGFP were exposed to 95% air/5% CO₂ (Ctl) or 2 vol% isoflurane in 95% air/5% CO₂ for 20 min at 37°C. At 20 min, isoflurane exposure was terminated and spine density and area was quantified at different time points (A, only 40 min wash shown). Changes in spine number and area observed after 20 min of isoflurane exposure recovered at 40 min of wash-out (B, C) by two-way ANOVA with Sidak post hoc test (*p<0.05; ****p<0.0001). Data are mean ± SEM; n = 30 to 40 dendritic ROIs per experimental group for spine density; n = 600 to 1800 dendritic spines per experimental group for spine area. Scale bar = 5 µm.