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Three-dimensional neurophenotyping of adult zebrafish behavior.

Cachat J, Stewart A, Utterback E, Hart P, Gaikwad S, Wong K, Kyzar E, Wu N, Kalueff AV - PLoS ONE (2011)

Bottom Line: The present large-scale study applied the newest video-tracking and data-mining technologies to further examine zebrafish anxiety-like phenotypes.It also enables rapid optimization of video tracking settings to improve quantification of automated parameters, and suggests that spatiotemporal organization of zebrafish swimming activity can be affected by various experimental manipulations in a manner predicted by their anxiolytic or anxiogenic nature.Our approach markedly enhances the power of zebrafish behavioral analyses, providing innovative framework for high-throughput 3D phenotyping of adult zebrafish behavior.

View Article: PubMed Central - PubMed

Affiliation: Department of Pharmacology and Neuroscience Program, Tulane Neurophenotyping Platform and Zebrafish Neuroscience Research Consortium, Tulane University Medical School, New Orleans, Louisiana, United States of America.

ABSTRACT
The use of adult zebrafish (Danio rerio) in neurobehavioral research is rapidly expanding. The present large-scale study applied the newest video-tracking and data-mining technologies to further examine zebrafish anxiety-like phenotypes. Here, we generated temporal and spatial three-dimensional (3D) reconstructions of zebrafish locomotion, globally assessed behavioral profiles evoked by several anxiogenic and anxiolytic manipulations, mapped individual endpoints to 3D reconstructions, and performed cluster analysis to reconfirm behavioral correlates of high- and low-anxiety states. The application of 3D swim path reconstructions consolidates behavioral data (while increasing data density) and provides a novel way to examine and represent zebrafish behavior. It also enables rapid optimization of video tracking settings to improve quantification of automated parameters, and suggests that spatiotemporal organization of zebrafish swimming activity can be affected by various experimental manipulations in a manner predicted by their anxiolytic or anxiogenic nature. Our approach markedly enhances the power of zebrafish behavioral analyses, providing innovative framework for high-throughput 3D phenotyping of adult zebrafish behavior.

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Flowchart illustrating the experimental strategy of this study.The rationale (A) includes examining traditional, manually recorded novel tank test behavioral endpoints across several treatments and trials (Step 1). Video-tracking analysis was then performed to generate additional automated behavioral endpoints and raw spatiotemporal data for three-dimensional (3D) swim path reconstructions (Steps 2–3), followed by hierarchical clustering (Step 4) across all behavioral endpoints and experimental treatments in order to discover potential overlaps between manual and automated endpoints. These overlaps were reconfirmed using the 3D swim path reconstructions (Step 5). Finally, our interpretation of the observed affective states was verified with measured endocrine responses (Step 6). The experimental process (B) was standardized for all novel tank trials. Naïve, wild-type zebrafish were placed in an unfamiliar, novel tank for 6 min. Animal behavior was manually observed and two cameras recorded videos for automated analysis in EthoVision XT7 (during which manual, event-based scoring was also performed). Track data for each subject was exported, processed and visualized in a 3D scatter plot with RapidMiner 5.0.
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pone-0017597-g001: Flowchart illustrating the experimental strategy of this study.The rationale (A) includes examining traditional, manually recorded novel tank test behavioral endpoints across several treatments and trials (Step 1). Video-tracking analysis was then performed to generate additional automated behavioral endpoints and raw spatiotemporal data for three-dimensional (3D) swim path reconstructions (Steps 2–3), followed by hierarchical clustering (Step 4) across all behavioral endpoints and experimental treatments in order to discover potential overlaps between manual and automated endpoints. These overlaps were reconfirmed using the 3D swim path reconstructions (Step 5). Finally, our interpretation of the observed affective states was verified with measured endocrine responses (Step 6). The experimental process (B) was standardized for all novel tank trials. Naïve, wild-type zebrafish were placed in an unfamiliar, novel tank for 6 min. Animal behavior was manually observed and two cameras recorded videos for automated analysis in EthoVision XT7 (during which manual, event-based scoring was also performed). Track data for each subject was exported, processed and visualized in a 3D scatter plot with RapidMiner 5.0.

Mentions: Recent studies have characterized adult zebrafish behavior in several novelty-based paradigms, reporting habituation [35], thigmotaxis, geotaxis and scototaxis [28], [34], [36], [37]. As a relatively young field, adult zebrafish behavioral neuroscience continues to adapt traditional rodent paradigms (such as open field, light-dark box, startle, and predator exposure tests) to the use in this aquatic species [4], [5], [31], [37], [38], [39], [40], [41]. Similar to rodent open field test [42], [43], [44], the novel tank test (Fig. 1 and 2) evaluates the natural neophobic response of zebrafish, expressed in reduced exploration, increased freezing and/or unorganized erratic locomotion [27], [28], [35], [39], [45]. In contrast, reduced anxiety in this test is accompanied by increased exploration with reduced freezing and fewer erratic bouts [28], [46] (Fig. 3).


Three-dimensional neurophenotyping of adult zebrafish behavior.

Cachat J, Stewart A, Utterback E, Hart P, Gaikwad S, Wong K, Kyzar E, Wu N, Kalueff AV - PLoS ONE (2011)

Flowchart illustrating the experimental strategy of this study.The rationale (A) includes examining traditional, manually recorded novel tank test behavioral endpoints across several treatments and trials (Step 1). Video-tracking analysis was then performed to generate additional automated behavioral endpoints and raw spatiotemporal data for three-dimensional (3D) swim path reconstructions (Steps 2–3), followed by hierarchical clustering (Step 4) across all behavioral endpoints and experimental treatments in order to discover potential overlaps between manual and automated endpoints. These overlaps were reconfirmed using the 3D swim path reconstructions (Step 5). Finally, our interpretation of the observed affective states was verified with measured endocrine responses (Step 6). The experimental process (B) was standardized for all novel tank trials. Naïve, wild-type zebrafish were placed in an unfamiliar, novel tank for 6 min. Animal behavior was manually observed and two cameras recorded videos for automated analysis in EthoVision XT7 (during which manual, event-based scoring was also performed). Track data for each subject was exported, processed and visualized in a 3D scatter plot with RapidMiner 5.0.
© Copyright Policy
Related In: Results  -  Collection

Show All Figures
getmorefigures.php?uid=PMC3049776&req=5

pone-0017597-g001: Flowchart illustrating the experimental strategy of this study.The rationale (A) includes examining traditional, manually recorded novel tank test behavioral endpoints across several treatments and trials (Step 1). Video-tracking analysis was then performed to generate additional automated behavioral endpoints and raw spatiotemporal data for three-dimensional (3D) swim path reconstructions (Steps 2–3), followed by hierarchical clustering (Step 4) across all behavioral endpoints and experimental treatments in order to discover potential overlaps between manual and automated endpoints. These overlaps were reconfirmed using the 3D swim path reconstructions (Step 5). Finally, our interpretation of the observed affective states was verified with measured endocrine responses (Step 6). The experimental process (B) was standardized for all novel tank trials. Naïve, wild-type zebrafish were placed in an unfamiliar, novel tank for 6 min. Animal behavior was manually observed and two cameras recorded videos for automated analysis in EthoVision XT7 (during which manual, event-based scoring was also performed). Track data for each subject was exported, processed and visualized in a 3D scatter plot with RapidMiner 5.0.
Mentions: Recent studies have characterized adult zebrafish behavior in several novelty-based paradigms, reporting habituation [35], thigmotaxis, geotaxis and scototaxis [28], [34], [36], [37]. As a relatively young field, adult zebrafish behavioral neuroscience continues to adapt traditional rodent paradigms (such as open field, light-dark box, startle, and predator exposure tests) to the use in this aquatic species [4], [5], [31], [37], [38], [39], [40], [41]. Similar to rodent open field test [42], [43], [44], the novel tank test (Fig. 1 and 2) evaluates the natural neophobic response of zebrafish, expressed in reduced exploration, increased freezing and/or unorganized erratic locomotion [27], [28], [35], [39], [45]. In contrast, reduced anxiety in this test is accompanied by increased exploration with reduced freezing and fewer erratic bouts [28], [46] (Fig. 3).

Bottom Line: The present large-scale study applied the newest video-tracking and data-mining technologies to further examine zebrafish anxiety-like phenotypes.It also enables rapid optimization of video tracking settings to improve quantification of automated parameters, and suggests that spatiotemporal organization of zebrafish swimming activity can be affected by various experimental manipulations in a manner predicted by their anxiolytic or anxiogenic nature.Our approach markedly enhances the power of zebrafish behavioral analyses, providing innovative framework for high-throughput 3D phenotyping of adult zebrafish behavior.

View Article: PubMed Central - PubMed

Affiliation: Department of Pharmacology and Neuroscience Program, Tulane Neurophenotyping Platform and Zebrafish Neuroscience Research Consortium, Tulane University Medical School, New Orleans, Louisiana, United States of America.

ABSTRACT
The use of adult zebrafish (Danio rerio) in neurobehavioral research is rapidly expanding. The present large-scale study applied the newest video-tracking and data-mining technologies to further examine zebrafish anxiety-like phenotypes. Here, we generated temporal and spatial three-dimensional (3D) reconstructions of zebrafish locomotion, globally assessed behavioral profiles evoked by several anxiogenic and anxiolytic manipulations, mapped individual endpoints to 3D reconstructions, and performed cluster analysis to reconfirm behavioral correlates of high- and low-anxiety states. The application of 3D swim path reconstructions consolidates behavioral data (while increasing data density) and provides a novel way to examine and represent zebrafish behavior. It also enables rapid optimization of video tracking settings to improve quantification of automated parameters, and suggests that spatiotemporal organization of zebrafish swimming activity can be affected by various experimental manipulations in a manner predicted by their anxiolytic or anxiogenic nature. Our approach markedly enhances the power of zebrafish behavioral analyses, providing innovative framework for high-throughput 3D phenotyping of adult zebrafish behavior.

Show MeSH