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Neurochemical measurements in the zebrafish brain.

Jones LJ, McCutcheon JE, Young AM, Norton WH - Front Behav Neurosci (2015)

Bottom Line: In this study we have used in vitro FSCV to measure the release of analytes in the adult zebrafish telencephalon.We compare different stimulation methods and present a characterization of neurochemical changes in the wild-type zebrafish brain.This study represents the first FSCV recordings in zebrafish, thus paving the way for neurochemical analysis of the fish brain.

View Article: PubMed Central - PubMed

Affiliation: Department of Neuroscience, Psychology and Behaviour, University of Leicester Leicester, UK.

ABSTRACT
The zebrafish is an ideal model organism for behavioral genetics and neuroscience. The high conservation of genes and neurotransmitter pathways between zebrafish and other vertebrates permits the translation of research between species. Zebrafish behavior can be studied at both larval and adult stages and recent research has begun to establish zebrafish models for human disease. Fast scan cyclic voltammetry (FSCV) is an electrochemical technique that permits the detection of neurotransmitter release and reuptake. In this study we have used in vitro FSCV to measure the release of analytes in the adult zebrafish telencephalon. We compare different stimulation methods and present a characterization of neurochemical changes in the wild-type zebrafish brain. This study represents the first FSCV recordings in zebrafish, thus paving the way for neurochemical analysis of the fish brain.

No MeSH data available.


Related in: MedlinePlus

Comparison of cyclic voltammograms generated by exposing electrodes to dopamine, 5-HT, histamine, and pH changes in a flow cell. Color plots (A,C,E,G,I,K) and cyclic voltammograms (B,D,F,H,J,L; black lines represent forward scan and red lines reverse scan) taken at the time point indicated by the dashed white lines. (A,B) 1 μM dopamine solution. (C,D) 0.5 μM 5-HT solution. (E,F) 40 μM histamine solution. (G,H) −0.25 units acidic pH change (pH 7.4 → pH 7.15). (I,J) -1.0 units acidic pH change (pH 7.4 → pH 6.4). (K,L) +1.0 units basic pH change (pH 7.4 → pH 7.4 → pH 8.4). Scale bar in (A,C,E,G,I,K) represents 5 s.
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Figure 2: Comparison of cyclic voltammograms generated by exposing electrodes to dopamine, 5-HT, histamine, and pH changes in a flow cell. Color plots (A,C,E,G,I,K) and cyclic voltammograms (B,D,F,H,J,L; black lines represent forward scan and red lines reverse scan) taken at the time point indicated by the dashed white lines. (A,B) 1 μM dopamine solution. (C,D) 0.5 μM 5-HT solution. (E,F) 40 μM histamine solution. (G,H) −0.25 units acidic pH change (pH 7.4 → pH 7.15). (I,J) -1.0 units acidic pH change (pH 7.4 → pH 6.4). (K,L) +1.0 units basic pH change (pH 7.4 → pH 7.4 → pH 8.4). Scale bar in (A,C,E,G,I,K) represents 5 s.

Mentions: As a first step toward characterizing analyte release in zebrafish we collected template FSCV data for dopamine, 5-HT and histamine, neurotransmitter systems that send extensive projections to the telencephalon. We used a flow cell (Sinkala et al., 2012), a microfluidic device that permits electrodes to be exposed to standard neurotransmitter solutions, to collect representative color plots and cyclic voltammograms. We exposed carbon fiber electrodes to a known concentration of each neurotransmitter or pH shift (using the voltage waveform shown in Figure 3B). Application of 1 μM dopamine produced an increase in current at ~ +0.6 V and a reduction peak at ~ −0.25 V (Figures 2A,B). A 0.5 μM 5-HT solution also produced an increase in current ~ +0.6 V but reduction peaks occurred at ~ 0 and ~ −0.5 V (Figures 2C,D). Application of 40 μM histamine produced a very different response, with an increase in current on the reverse scan at ~ +1.0 V and two reduction peaks on the forward scan at ~ +0.25 and ~ −0.4 V (Figures 2E,F). We also investigated the effect of altering pH on the oxidation potentials of voltammograms. An acidic change of −0.25 pH units (i.e., pH 7.15) produced an oxidation peak at ~ +0.5 V and reduction peaks at both ~ +1.1 and ~ −0.3 V (Figures 2G,H). An acidic change of −1.0 pH units (i.e., pH 6.4) produced a similar voltammogram with a sharp oxidation peak at ~ +0.5 V and reduction peaks at both ~ +1.1 and ~ −0.3 V (Figures 2I,J). A basic pH change of +1.0 units (i.e., pH 8.4) produced a large oxidation peak on the reverse scan at ~ +1.1 V and reduction peaks on the forward scans at ~ +0.4 and ~ −0.5 V, similar to the neurotransmitter histamine (Figures 2K,L). Together, these experiments demonstrate the characteristic shapes of cyclic voltammograms that are produced by exposing electrodes to neurotransmitter solutions and changes in pH.


Neurochemical measurements in the zebrafish brain.

Jones LJ, McCutcheon JE, Young AM, Norton WH - Front Behav Neurosci (2015)

Comparison of cyclic voltammograms generated by exposing electrodes to dopamine, 5-HT, histamine, and pH changes in a flow cell. Color plots (A,C,E,G,I,K) and cyclic voltammograms (B,D,F,H,J,L; black lines represent forward scan and red lines reverse scan) taken at the time point indicated by the dashed white lines. (A,B) 1 μM dopamine solution. (C,D) 0.5 μM 5-HT solution. (E,F) 40 μM histamine solution. (G,H) −0.25 units acidic pH change (pH 7.4 → pH 7.15). (I,J) -1.0 units acidic pH change (pH 7.4 → pH 6.4). (K,L) +1.0 units basic pH change (pH 7.4 → pH 7.4 → pH 8.4). Scale bar in (A,C,E,G,I,K) represents 5 s.
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Figure 2: Comparison of cyclic voltammograms generated by exposing electrodes to dopamine, 5-HT, histamine, and pH changes in a flow cell. Color plots (A,C,E,G,I,K) and cyclic voltammograms (B,D,F,H,J,L; black lines represent forward scan and red lines reverse scan) taken at the time point indicated by the dashed white lines. (A,B) 1 μM dopamine solution. (C,D) 0.5 μM 5-HT solution. (E,F) 40 μM histamine solution. (G,H) −0.25 units acidic pH change (pH 7.4 → pH 7.15). (I,J) -1.0 units acidic pH change (pH 7.4 → pH 6.4). (K,L) +1.0 units basic pH change (pH 7.4 → pH 7.4 → pH 8.4). Scale bar in (A,C,E,G,I,K) represents 5 s.
Mentions: As a first step toward characterizing analyte release in zebrafish we collected template FSCV data for dopamine, 5-HT and histamine, neurotransmitter systems that send extensive projections to the telencephalon. We used a flow cell (Sinkala et al., 2012), a microfluidic device that permits electrodes to be exposed to standard neurotransmitter solutions, to collect representative color plots and cyclic voltammograms. We exposed carbon fiber electrodes to a known concentration of each neurotransmitter or pH shift (using the voltage waveform shown in Figure 3B). Application of 1 μM dopamine produced an increase in current at ~ +0.6 V and a reduction peak at ~ −0.25 V (Figures 2A,B). A 0.5 μM 5-HT solution also produced an increase in current ~ +0.6 V but reduction peaks occurred at ~ 0 and ~ −0.5 V (Figures 2C,D). Application of 40 μM histamine produced a very different response, with an increase in current on the reverse scan at ~ +1.0 V and two reduction peaks on the forward scan at ~ +0.25 and ~ −0.4 V (Figures 2E,F). We also investigated the effect of altering pH on the oxidation potentials of voltammograms. An acidic change of −0.25 pH units (i.e., pH 7.15) produced an oxidation peak at ~ +0.5 V and reduction peaks at both ~ +1.1 and ~ −0.3 V (Figures 2G,H). An acidic change of −1.0 pH units (i.e., pH 6.4) produced a similar voltammogram with a sharp oxidation peak at ~ +0.5 V and reduction peaks at both ~ +1.1 and ~ −0.3 V (Figures 2I,J). A basic pH change of +1.0 units (i.e., pH 8.4) produced a large oxidation peak on the reverse scan at ~ +1.1 V and reduction peaks on the forward scans at ~ +0.4 and ~ −0.5 V, similar to the neurotransmitter histamine (Figures 2K,L). Together, these experiments demonstrate the characteristic shapes of cyclic voltammograms that are produced by exposing electrodes to neurotransmitter solutions and changes in pH.

Bottom Line: In this study we have used in vitro FSCV to measure the release of analytes in the adult zebrafish telencephalon.We compare different stimulation methods and present a characterization of neurochemical changes in the wild-type zebrafish brain.This study represents the first FSCV recordings in zebrafish, thus paving the way for neurochemical analysis of the fish brain.

View Article: PubMed Central - PubMed

Affiliation: Department of Neuroscience, Psychology and Behaviour, University of Leicester Leicester, UK.

ABSTRACT
The zebrafish is an ideal model organism for behavioral genetics and neuroscience. The high conservation of genes and neurotransmitter pathways between zebrafish and other vertebrates permits the translation of research between species. Zebrafish behavior can be studied at both larval and adult stages and recent research has begun to establish zebrafish models for human disease. Fast scan cyclic voltammetry (FSCV) is an electrochemical technique that permits the detection of neurotransmitter release and reuptake. In this study we have used in vitro FSCV to measure the release of analytes in the adult zebrafish telencephalon. We compare different stimulation methods and present a characterization of neurochemical changes in the wild-type zebrafish brain. This study represents the first FSCV recordings in zebrafish, thus paving the way for neurochemical analysis of the fish brain.

No MeSH data available.


Related in: MedlinePlus