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Accumulation of an Antidepressant in Vesiculogenic Membranes of Yeast Cells Triggers Autophagy

Figure 1

[3H]sertraline cellular uptake and accumulation is rapid, partially lysosomotropic and varies from cell to cell.

(A) Wildtype BY4716 cells accumulate [3H]sertraline (picomoles of radioligand/107cells) in a time-dependent manner as a function of ambient pH: pH 6.0 (diamonds); pH 6.5 (triangles); pH 7.0 (squares). Notice the break in the x-axis between 10 and 20. (B) Bar graph showing fraction of [3H]sertraline accumulated with a 30-minute 2 µM bafilomycin A pre-treatment (“before”) compared to no bafilomycin A pre-treatment; and fraction of [3H]sertraline accumulated with a 30-minute 2 µM bafilomycin A-post-treatment (“after”) compared to no bafilomycin A post-treatment. Wildtype BY4716 (black, unfilled) is contrasted with a vma9 mutant (black, filled). Before and after treatments of wildtype BY4716 treated with 1 µM FCCP (blue, unfilled), 1 µM FCCP+2 µM bafilomycin A (blue, filled), or 10 µg/mL oligomycin (red, unfilled) are also shown. Statistical significance determined by two-tailed t-test. *** P = 0.0004; * P = 0.0118. (C) [3H]sertraline uptake and accumulation time course with wildtype BY4742 cells (black circles, filled) a dnf1Δ dnf2Δ dnf3Δ triple mutant cells (red circles, filled), and a vma9 mutant (red circles, unfilled). Notice the break in the y-axis between 0.02 and 0.05. (D) Single-cell dose response of sertraline-induced cytotoxicity (“overdose”) for wildtype BY4716 (black), vma9 (red), and cup5 (cyan) cells. Error bars indicate SEM. Means were generated from three independent biological replicates.

Figure 1

doi: https://doi.org/10.1371/journal.pone.0034024.g001