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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.


Comparison of analytes evoked by high K+ aCSF stimulation of the zebrafish telencephalon. (A) Color plot showing changes in current following stimulation with high K+ aCSF. (B) Applied waveform ranging from 0 V → +1.2 V → −0.6 V → 0 V (the holding potential). The forward scan of the waveform is colored black and the reverse scan is red. (C) Current vs. time plot showing the profile of current changes at Eapp = ~ +0.6 V, the point in the waveform indicated by the black circle in (E). (D) Current vs. time plot showing the profile of current changes at Eapp = ~ +1.0 V on the reverse scan of the waveform, the point indicated by the circle in (F). (E) Representative cyclic voltammogram taken at the time point indicated by the thick dashed white line in (A). (F) Representative cyclic voltammogram taken at the time point indicated by the thin dashed white line in (A). (G) Cyclic voltammograms from 24 separate stimulations taken at the time point indicated by the thick dashed white line in (A). (H) Cyclic voltammograms from 20 independent experiments taken at the time point indicated by the thin dashed white line in (A). (I) Average cyclic voltammogram derived from data presented in (G). (J) Average cyclic voltammogram derived from data presented in (H).
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Figure 3: Comparison of analytes evoked by high K+ aCSF stimulation of the zebrafish telencephalon. (A) Color plot showing changes in current following stimulation with high K+ aCSF. (B) Applied waveform ranging from 0 V → +1.2 V → −0.6 V → 0 V (the holding potential). The forward scan of the waveform is colored black and the reverse scan is red. (C) Current vs. time plot showing the profile of current changes at Eapp = ~ +0.6 V, the point in the waveform indicated by the black circle in (E). (D) Current vs. time plot showing the profile of current changes at Eapp = ~ +1.0 V on the reverse scan of the waveform, the point indicated by the circle in (F). (E) Representative cyclic voltammogram taken at the time point indicated by the thick dashed white line in (A). (F) Representative cyclic voltammogram taken at the time point indicated by the thin dashed white line in (A). (G) Cyclic voltammograms from 24 separate stimulations taken at the time point indicated by the thick dashed white line in (A). (H) Cyclic voltammograms from 20 independent experiments taken at the time point indicated by the thin dashed white line in (A). (I) Average cyclic voltammogram derived from data presented in (G). (J) Average cyclic voltammogram derived from data presented in (H).

Mentions: We next investigated whether fast-scan cyclic voltammetry (FSCV) could be used to measure the release of analytes in sagittal sections of the adult zebrafish brain. The zebrafish dorsal telencephalon receives extensive 5-HT-positive projections from the raphe- and pretectal nuclei (Lillesaar et al., 2009). We therefore applied a voltage waveform optimized for measurements of 5-HT (John and Jones, 2007a) and depolarised neurons and terminals with aCSF containing a high concentration of K+ (100 mM K+; hereafter high K+ aCSF). Bath application of high K+ aCSF led to changes in current at several points in the voltage waveform. A cyclic voltammogram extracted at ~10 s after stimulation displays characteristics that could reflect the oxidation of dopamine and/or 5-HT, including a prominent peak in current on the forward scan at ~ +0.6 V (Figure 3E). A rapid increase in oxidative current is observed at the point in the waveform that corresponds to the peak of this signal (~ +0.6 V; Figure 3C). However, this current vs. time plot also exhibits a striking dip in current which is most likely due to the decrease in current at around ~ +0.2 V masking the oxidation peak at ~ +0.6 V.


Neurochemical measurements in the zebrafish brain.

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

Comparison of analytes evoked by high K+ aCSF stimulation of the zebrafish telencephalon. (A) Color plot showing changes in current following stimulation with high K+ aCSF. (B) Applied waveform ranging from 0 V → +1.2 V → −0.6 V → 0 V (the holding potential). The forward scan of the waveform is colored black and the reverse scan is red. (C) Current vs. time plot showing the profile of current changes at Eapp = ~ +0.6 V, the point in the waveform indicated by the black circle in (E). (D) Current vs. time plot showing the profile of current changes at Eapp = ~ +1.0 V on the reverse scan of the waveform, the point indicated by the circle in (F). (E) Representative cyclic voltammogram taken at the time point indicated by the thick dashed white line in (A). (F) Representative cyclic voltammogram taken at the time point indicated by the thin dashed white line in (A). (G) Cyclic voltammograms from 24 separate stimulations taken at the time point indicated by the thick dashed white line in (A). (H) Cyclic voltammograms from 20 independent experiments taken at the time point indicated by the thin dashed white line in (A). (I) Average cyclic voltammogram derived from data presented in (G). (J) Average cyclic voltammogram derived from data presented in (H).
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Figure 3: Comparison of analytes evoked by high K+ aCSF stimulation of the zebrafish telencephalon. (A) Color plot showing changes in current following stimulation with high K+ aCSF. (B) Applied waveform ranging from 0 V → +1.2 V → −0.6 V → 0 V (the holding potential). The forward scan of the waveform is colored black and the reverse scan is red. (C) Current vs. time plot showing the profile of current changes at Eapp = ~ +0.6 V, the point in the waveform indicated by the black circle in (E). (D) Current vs. time plot showing the profile of current changes at Eapp = ~ +1.0 V on the reverse scan of the waveform, the point indicated by the circle in (F). (E) Representative cyclic voltammogram taken at the time point indicated by the thick dashed white line in (A). (F) Representative cyclic voltammogram taken at the time point indicated by the thin dashed white line in (A). (G) Cyclic voltammograms from 24 separate stimulations taken at the time point indicated by the thick dashed white line in (A). (H) Cyclic voltammograms from 20 independent experiments taken at the time point indicated by the thin dashed white line in (A). (I) Average cyclic voltammogram derived from data presented in (G). (J) Average cyclic voltammogram derived from data presented in (H).
Mentions: We next investigated whether fast-scan cyclic voltammetry (FSCV) could be used to measure the release of analytes in sagittal sections of the adult zebrafish brain. The zebrafish dorsal telencephalon receives extensive 5-HT-positive projections from the raphe- and pretectal nuclei (Lillesaar et al., 2009). We therefore applied a voltage waveform optimized for measurements of 5-HT (John and Jones, 2007a) and depolarised neurons and terminals with aCSF containing a high concentration of K+ (100 mM K+; hereafter high K+ aCSF). Bath application of high K+ aCSF led to changes in current at several points in the voltage waveform. A cyclic voltammogram extracted at ~10 s after stimulation displays characteristics that could reflect the oxidation of dopamine and/or 5-HT, including a prominent peak in current on the forward scan at ~ +0.6 V (Figure 3E). A rapid increase in oxidative current is observed at the point in the waveform that corresponds to the peak of this signal (~ +0.6 V; Figure 3C). However, this current vs. time plot also exhibits a striking dip in current which is most likely due to the decrease in current at around ~ +0.2 V masking the oxidation peak at ~ +0.6 V.

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.