7 research outputs found

    Dopamine release by optogenetic stimulation.

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    <p>(A) Extracellular dopamine level recording with blue light illumination for 10 seconds on EGFP- and ChR2-EGFP-expressing PC12 cells. (C) ChR2-mediated dopamine release under different illumination durations, from 5 seconds to 20 seconds. Arrow indicates the start of illumination. (A) and (C) are dopamine signals in current vs. time. (B) and (D) are integrations of evoked currents with time during blue light illumination. Each column represents mean ± SEM from five independent experiments. ***, <i>P</i><0.001.</p

    The <i>in vitro</i> real-time dopamine recording system.

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    <p>(A) Schematic diagram of working electrode (WE), counter electrode (CE), and reference electrode (RE) for dopamine sensing of ChR2-transfected PC12 cells under optogenetic stimulation at 473 nm. (B) Fluorescence and (C) phase-contrast images of ChR2-EGFP-expressing PC12 cells and electrode (arrows) under wide-field fluorescence microscope.</p

    Extracellular calcium influx during optogenetic stimulation.

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    <p>(A) Relative intracellular Ca<sup>2+</sup> levels of ratiometric fura-2 fluorescence intensity (ratio 340/380) for EGFP- and ChR2-EGFP-expressing PC12 cells in 1.8 mM Ca<sup>2+</sup> buffer with or without 10 µM Nifedipine, or Ca<sup>2+</sup>-free buffer under illumination of blue light for 10 seconds. The Ca<sup>2+</sup> signals during light illumination that could not be read out are depicted as dotted lines. (B) The changes of relative intracellular Ca<sup>2+</sup> levels (mean ± SEM) by the fura-2 ratiometric imaging (Δ ratio 340/380) from at least 50 cells of four independent experiments. ***, <i>P</i><0.001.</p

    Effect of calcium influx on ChR2-mediated dopamine release.

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    <p>(A) Simultaneous optical stimulation, dopamine current recording and Ca<sup>2+</sup> imaging. Blue light illumination on ChR2-EGFP-expressing cells in controlled 1.8 mM Ca<sup>2+</sup> buffer without or with 10 µM Nifedipine, and Ca<sup>2+</sup>-free buffer. The light-related artifact is not removed. (B) Significant inhibition in dopamine current during blue light illumination in Ca<sup>2+</sup>-free and Ca<sup>2+</sup> buffer with Nifedipine. Each column represents mean ± SEM from five independent experiments. ***<i>P</i><0.001.</p

    Characterization of Au-NP/SAM modified electrodes.

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    <p>(A) CV curves of 10 µM dopamine recorded by native (-Δ-) and Au-NP/SAM (-•-) platinum microelectrodes at a scan rate of 10 V s<sup>−1</sup>. The inserted picture represents the CV curve after background subtraction. SEM images showing the surface morphologies of (B) native and (C) Au-NP/SAM platinum electrodes. (D) Amperometric <i>i-t</i> curves of Au-NP/SAM microelectrodes recorded in PBS for dopamine calibration at a scan potential of 0.25 V (<i>vs.</i> Ag/AgCl).</p

    Effect of stimulation parameters of blue light on calcium influx and dopamine release.

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    <p>Optogenetic stimulation at frequencies of 10, 20, and 30(CI) of blue laser light were applied for 10 seconds. (A) Changes of relative intracellular Ca<sup>2+</sup> levels by the fura-2 ratiometric imaging (Δ ratio 340/380). Each column represents mean ± SEM from at least 60 cells of three independent experiments. *: significant difference within the same group; #: significant difference between groups. # and *, <i>P</i><0.05; **, <i>P</i><0.01. (B) Background current of stimulatory artifacts and (C) dopamine release under 10 Hz/10 ms, 30 Hz/30 ms, and CI of blue light.</p
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