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

FIGURE 6.

FIGURE 6

Inhibition of synaptic vesicle exocytosis by isoflurane is acutely irreversible in Val66Met and Met66Met neurons, but rescued with exogenous BDNF. Mouse hippocampal neuron cultures (16DIV) transfected with vGlut-pH and mCherry were perfused with control buffer, 0.5–0.7 mM isoflurane, or mBDNF (75–100 ng/ml) with tetanic stimulation (lightning bolt) as indicated in (A,B, top). Tetanic stimulation of synaptic vesicle exocytosis measured with vGlut-pH was inhibited by isoflurane (0.5–0.7 mM) in Val66Val mouse neurons compared to Val66Met and Met66Met neurons (**p < 0.01 by multiple comparisons one-way ANOVA with Dunnett’s post-hoc test). Changes in vesicle exocytosis observed in Val66Met and Met66Met neurons after isoflurane exposure did not recover following washout of drug (A, bottom) (*p < 0.05 by multiple comparisons one-way ANOVA with Dunnett’s post-hoc test). Addition of 75–100 ng/ml of exogenous mBDNF during washout rescued synaptic vesicle exocytosis in Val66Met and Met66Met and increased exocytosis in Val66Val (B, bottom) (*p < 0.05; **p < 0.01 neurons by Student’s unpaired t-test). Data are mean±SD; n = 4–14 neurons per experimental group and 200–700 boutons.