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. 2015 Jan 29;29(5):2112–2119. doi: 10.1096/fj.14-267179

Figure 2.

Figure 2.

Bumblebee brain neurons undergo mitochondrial depolarization when nAChR are hyperstimulated. A) Bumblebee neurons in culture undergo mitochondrial depolarization in the presence of high levels (1 mM; open circles) but not low levels (100 μM; filled circles) of acetylcholine. The neonicotinoid, clothianidin (B), induces mitochondrial depolarization at 10 nM (open circles) but not at 1 nM (filled circles), and imidacloprid (C) induces mitochondrial depolarization at 1 µM (open circles) but not at 10 nM (filled circles). (D) Neurons pre-exposed to the nAChR antagonist d-tubucurarine (500 μM) do not undergo mitochondrial depolarization in the presence of clothianidin (100 nM), demonstrating an nAChR-dependent process. In all cases, mitochondrial depolarization was monitored using ratiometric (red/green) JC-1 imaging, and the experiment was terminated by full mitochondrial depolarization using 2,4-dinitophenol (1 mM). In all cases, 15–20 regions of interest were monitored (n = 3).