Voltammetry: Electrochemical Detection of Neurotransmitters in the Brain

Abstract

Voltammetry is an electrochemical technique that capitalises on the ability of some substances to become oxidised or reduced. A variety of voltammetric methods have been developed for the detection of biogenic amines such as dopamine, noradrenaline and serotonin in the brain. Each method differs in selectivity for the transmitter of interest and in temporal resolution. Of these, fast‐scan cyclic voltammetry (FCV) at carbon fibre electrodes has been used extensively for monitoring the evoked or spontaneous release of biogenic amines in various brain regions with temporal and spatial resolutions that capture extrasynaptic transmission. Studies in rodent and non‐rodent brain slices containing either monoamine cell bodies or axonal projections enable the dynamics of neurotransmitter release and its regulation by monoamine transporters, autoreceptors and local neuromodulators to be examined. Furthermore, rapid detection of release in freely moving animals can reveal the role of biogenic amines in motivated behaviour.

Key Concepts

  • Dopamine, noradrenaline and serotonin are the major biogenic amine transmitters in the brain.
  • Voltammetry is an electrochemical method that detects the current generated when an electroactive molecule is oxidised; the current generated is directly proportional to the number of molecules oxidised.
  • Many biologically active substances are electroactive and therefore possess the ability to become oxidised or reduced when a sufficient voltage is applied at a suitable electrochemical electrode.
  • Carbon fibre microelectrodes are excellent electrochemical sensors for monitoring biogenic amines in the brain.
  • Fast‐scan cyclic voltammetry (FCV) at carbon fibre microelectrode sensors resolves electroactive compounds by way of their peak oxidation and/or reduction potentials, and is the most popular voltammetric method used to detect neurotransmitter release.
  • Studies of biogenic amine release regulation using FCV have made significant contributions to our present understanding of neurotransmitter release regulation and the role of monoamines in normal and pathological conditions.
  • Brain slices have been used extensively for studies of monoamine release regulation using voltammetric methods.

Keywords: fast‐scan cyclic voltammetry; amperometry; carbon fibre microelectrodes; dopamine; serotonin; noradrenaline; neurotransmitter release; brain slices; oxidation potential; striatum

Figure 1. Redox reactions for catechols and 5‐hydroxyindoles. (a). Oxidation of catechols, such as dopamine (DA), noradrenaline (NA) or their metabolite dihydroxyphenylacetic acid (DOPAC), releases electrons and forms a catechol ortho‐quinone. (b). Oxidation of 5‐hydroxyindoles, such as serotonin (also known as 5‐hydroxytryptamine, 5‐HT), 5‐hydroxyindoleacetic acid (5‐HIAA) or 5‐hydroxytryptophan (5‐HTP), releases electrons and forms a quinoneimine. (c). Diagram showing the tip of a carbon fibre microelectrode at which catechols and 5‐hydroxyindoles are oxidised when a suitable potential is applied. Catechols are easier to oxidise than indoles because electrons are more readily removed from hydroxyl groups than from NH groups. The current generated by the released electrons is detected by the same carbon fibre electrode. The carbon fibre is typically only 7 µm in diameter, allowing monoamine transmission to be monitored in ‘real space’.
Figure 2. Types of voltammetric waveforms used to detect brain monoamine release. Amperometry involves holding a constant potential at an electrode which is sufficient to oxidise the electroactive species. Linear sweep voltammetry (LSV) applies a slow voltage ramp to the electrode which sequentially oxidises multiple electroactive analytes. Chronoamperometry uses the repetitive application of brief voltage pulses from an initial resting potential to a potential sufficient to oxidise the species of interest. This improves temporal resolution but lacks signal identity. Fast‐scan cyclic voltammetry (FCV) uses a triangular, cyclic, waveform with a scan rate of >300 V s−1. This enables electroactive substances to be sequentially oxidised and reduced which aids signal identification. The waveform is typically applied 10 times a second, allowing monoamine transmission to be monitored in ‘real time’.
Figure 3. Methodology for the detection of monoamines using fast‐scan cyclic voltammetry. (a) Triangular voltage waveform applied at a carbon fibre microelectrode up to 10 times a second. (b) Background charging current waveform generated at a carbon fibre microelectrode in response to the applied voltage in (a) obtained in the absence (black trace) and presence (red trace) of 1 μM dopamine. Oxidation (o) and reduction (r) peaks appear in the presence of dopamine. (c) Faradaic current waveform obtained by subtracting the background current (b) obtained in the absence from that in the presence of dopamine. The inset shows the Faradaic waveform expressed as a cyclic voltammogram. (d) Monitoring the current at the peak oxidation potential with successive voltammetric sweeps provides the time course of a dopamine release transient. (e) Records obtained in a striatal brain slice showing the time course of dopamine release evoked by a single‐pulse stimulation in the absence (red trace) and presence (blue trace) of a dopamine transporter (DAT) inhibitor. The peak extracellular dopamine concentration ([DA]o) is increased and the time course prolonged when dopamine uptake is impaired.
Figure 4. Rat brain sections showing major striatal and midbrain dopamine regions studied using FCV. (a) Cyclic voltammogram for 0.3 μM dopamine showing a single oxidation peak (o) and single reduction peak (r). (b) Diagram of a coronal striatal section that contains dopamine axons in the caudate putamen (CPu) within the dorsal striatum, as well as in the nucleus accumbens (NAc, core and shell) and in the olfactory tubercle (OTu) within the ventral striatum. (c) Diagram of a coronal midbrain section that contains dopamine cell bodies within the substantia nigra pars compacta (SNc‐coloured strip) and ventral tegmental area (VTA). The small brain insets indicate the level at which each coronal section lies within the brain (red lines). Below each coronal section are records of dopamine release versus time profiles obtained in different rat striatal or guinea‐pig midbrain subregions using FCV. Release was evoked by local electrical stimulation (30 pulses at 10 Hz) and expressed as extracellular dopamine concentration ([DA]o). Brain section diagrams are modified from Figures 20 and 73 of ‘Paxinos and Watson, ’. Reproduced and modified with permission from Paxinos et al., (2009) © Elsevier.
Figure 5. Rat brain sections showing some major noradrenaline and serotonin regions studied by FCV. (a) Cyclic voltammogram for 0.3 μM noradrenaline showing a single oxidation peak (o) and single reduction peak (r). (b, c) Diagram of coronal sections that contain noradrenaline cell bodies in the locus coeruleus (LC) or axons in the bed nucleus of the stria terminalis (BNST). (d) Cyclic voltammogram for 0.3 μM serotonin showing a single oxidation peak (o) and double reduction peaks (r1 and r2). (e, f) Diagram of coronal sections that contain serotonin cell bodies within the raphe nuclei (RN), including the dorsal raphe nucleus (top) and the median raphe nucleus (bottom) or serotonin axons in the superchiasmatic nucleus (SNC). The small brain insets indicate the level at which each coronal section lies within the brain (red lines). Brain section diagrams are modified from Figures 115, 36, 93 and 38 of ‘Paxinos and Watson, ’. Reproduced and modified with permission from Paxinos et al., (2009) © Elsevier.
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Further Reading

Adams R (1969) Electrochemistry at Solid Electrodes. New York: Dekker.

Boulton AA, Baker GB and Adams RN (eds) (1995) Voltammetric Methods in Brain Systems. Totowa, NJ: Humana.

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Michael AC and Borland LM (eds) (2007) Electrochemical Methods for Neuroscience. Boca Raton, FL: CRC Press.

Stamford JA (ed) (1992) Monitoring Neuronal Activity. Oxford: Oxford University Press.

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Patel, Jyoti C(Jan 2016) Voltammetry: Electrochemical Detection of Neurotransmitters in the Brain. In: eLS. John Wiley & Sons Ltd, Chichester. http://www.els.net [doi: 10.1002/9780470015902.a0025817]