Fast-scan cyclic voltammetry analysis of dynamic serotonin reponses to acute escitalopram.
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ABSTRACT:
SUBMITTER: Wood KM
PROVIDER: S-EPMC3656741 | biostudies-other | 2013 May
REPOSITORIES: biostudies-other
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Project description:Selective serotonin reuptake inhibitors (SSRIs) have been used for decades to treat disorders linked to serotonin dysregulation in the brain. Moreover, SSRIs are often used in studies aimed at measuring serotonin with fast-scan cyclic voltammetry (FSCV) in living tissues. Here, we show that three different SSRIs - fluoxetine, escitalopram, and sertraline - significantly diminish the faradaic oxidation current of serotonin when employing the commonly used Jackson waveform. Coating carbon-fiber microelectrodes (CFMs) with Nafion resulted in further degradation of peak current, increased response times, and decreased background charging currents compared to bare CFMs. To decrease fouling, we employed a recently published extended serotonin waveform, which scans to a maximum positive potential of +1.3 V, rather than +1.0 V used in the Jackson waveform. Use of this waveform with bare CFMs alleviated the decrease in faradaic current, indicating decreased electrode fouling. Collectively, our results suggest that fouling considerations are important when designing FSCV experiments that employ SSRIs and that they can be overcome by using the appropriate waveform.
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Project description:Serotonin is a neuromodulator implicated in depression that is often measured in real-time by fast-scan cyclic voltammetry (FSCV). A specialized "Jackson" waveform (JW, 0.2, 1.0 V, -0.1 V, 0.2 V, 1000 V s-1) was developed to reduce serotonin fouling, but the 1.0 V switching potential limits sensitivity and electrodes still foul. The goal of this study was to test the effects of extending the FSCV switching potential to increase serotonin sensitivity and decrease fouling. We compared the Jackson waveform, the dopamine waveform (DA, -0.4 V, 1.3 V, 400 V s-1), and two new waveforms: the extended serotonin waveform (ESW, 0.2, 1.3, -0.1, 0.2, 1000 V s-1) and extended hold serotonin waveform (EHSW, 0.2, 1.3 (hold 1 ms), -0.1, 0.2, 400 V s-1). The EHSW was the most sensitive (LOD = 0.6 nM), and the JW the least sensitive (LOD = 2.4 nM). With the Jackson waveform, electrode fouling was significant with repeated injections of serotonin or exposure to its metabolite, 5-hydroxyindoleacetic acid (5-HIAA). Using the extended waveforms, electrodes fouled 50% less than with the Jackson waveform for both analytes. No electrode fouling was observed with the dopamine waveform because of the negative holding potential. The Jackson waveform was the most selective for serotonin over dopamine (800×), and the ESW was also highly selective. All waveforms were useful for measuring serotonin with optogenetic stimulation in Drosophila larvae. These results provide new FSCV waveforms to measure dynamic serotonin changes with different experimental requirements, like high sensitivity (EHSW), high selectivity (ESW, JW), or eliminating electrode fouling (DA).
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