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. 2022 Jun 23;15:927149. doi: 10.3389/fnmol.2022.927149

FIGURE 1.

FIGURE 1

Isoflurane reversibly inhibits electrically-evoked synaptic vesicle exocytosis. Rat hippocampal neuron cultures (16DIV) transfected with vGlut-pH and mCherry were perfused with 0.5–0.7 mM isoflurane or control buffer with tetanic stimulation (lightning bolt) and imaged as indicated in (A). Cells were equilibrated with control buffer or buffer with isoflurane for 5 min prior to stimulation and time-lapse recording for 2 min. Representative images showing dendritic arbor and axons of a control neuron (B, mCherry). Inset shows single boutons (vGlut-pH) before (C, left) and after (C, right) tetanic stimulation with defined ROIs used for analysis (white circles). Representative paired traces of electrically-evoked synaptic vesicle exocytosis (black bar, tetanic stimulation) of ROIs (average of ∼50) from a single neuron under control, isoflurane, and washout conditions during 2 min recording (D, left). Peak synaptic vesicle exocytosis was reduced by isoflurane (D, right; peak fluorescence) (**p < 0.01 by one-way multiple comparisons ANOVA with Dunnett’s post-hoc test) and recovered following washout (D, right; peak fluorescence) (*p < 0.05 by one-way multiple comparisons ANOVA with Dunnett’s post-hoc test). Data are mean±SD%; n = 9 neurons per experimental group and 405–450 boutons. Scale bar = 5 μm.