TY - JOUR
T1 - Comparison between two- And three-dimensional scoring of zebrafish response to psychoactive drugs
T2 - Identifying when three-dimensional analysis is needed
AU - MacRì, Simone
AU - Clément, Romain J.G.
AU - Spinello, Chiara
AU - Porfiri, Maurizio
N1 - Funding Information:
The following grant information was disclosed by the authors: National Institutes of Health, National Institute on Drug Abuse: 1R21DA042558-01A1. Behavioral and Social Sciences Research that co-founded the National Institute on Drug Abuse. National Science Foundation: CMMI-1505832.
Funding Information:
This work was supported by the National Institutes of Health, National Institute on Drug Abuse under grant number 1R21DA042558-01A1, the Office of Behavioral and Social Sciences Research that co-funded the National Institute on Drug Abuse grant, and by the National Science Foundation under grant number CMMI-1505832. The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.
Funding Information:
Simultaneous recording from both cameras was initiated before transferring the fish into the test tank. In addition, at the beginning of the recording, a laser beam, visible from both cameras, was pointed into the test tank in order to ensure later synchronization of both video streams. At the end of the experiment, the fish was hand-netted into a separate tank. All the experiments were performed at the New York University Tandon School of Engineering (Brooklyn, NY, USA) in accordance with relevant guidelines and regulations, with National Institutes of Health guide for the care and use of Laboratory animals (NIH Publications No. 8023, revised 1978), and was approved by the University Animal Welfare Committee of New York University under protocol number 13-1424.
Publisher Copyright:
© 2019 Macrì et al.
PY - 2019
Y1 - 2019
N2 - Zebrafish (Danio rerio) have recently emerged as a valuable laboratory species in the field of behavioral pharmacology, where they afford rapid and precise high-throughput drug screening. Although the behavioral repertoire of this species manifests along three-dimensional (3D), most of the efforts in behavioral pharmacology rely on two-dimensional (2D) projections acquired from a single overhead or front camera. We recently showed that, compared to a 3D scoring approach, 2D analyses could lead to inaccurate claims regarding individual and social behavior of drug-free experimental subjects. Here, we examined whether this conclusion extended to the field of behavioral pharmacology by phenotyping adult zebrafish, acutely exposed to citalopram (30, 50, and 100 mg/L) or ethanol (0.25%, 0.50%, and 1.00%), in the novel tank diving test over a 6-min experimental session. We observed that both compounds modulated the time course of general locomotion and anxiety-related profiles, the latter being represented by specific behaviors (erratic movements and freezing) and avoidance of anxiety-eliciting areas of the test tank (top half and distance from the side walls). We observed that 2D projections of 3D trajectories (ground truth data) may introduce a source of unwanted variation in zebrafish behavioral phenotyping. Predictably, both 2D views underestimate absolute levels of general locomotion. Additionally, while data obtained from a camera positioned on top of the experimental tank are similar to those obtained from a 3D reconstruction, 2D front view data yield false negative findings.
AB - Zebrafish (Danio rerio) have recently emerged as a valuable laboratory species in the field of behavioral pharmacology, where they afford rapid and precise high-throughput drug screening. Although the behavioral repertoire of this species manifests along three-dimensional (3D), most of the efforts in behavioral pharmacology rely on two-dimensional (2D) projections acquired from a single overhead or front camera. We recently showed that, compared to a 3D scoring approach, 2D analyses could lead to inaccurate claims regarding individual and social behavior of drug-free experimental subjects. Here, we examined whether this conclusion extended to the field of behavioral pharmacology by phenotyping adult zebrafish, acutely exposed to citalopram (30, 50, and 100 mg/L) or ethanol (0.25%, 0.50%, and 1.00%), in the novel tank diving test over a 6-min experimental session. We observed that both compounds modulated the time course of general locomotion and anxiety-related profiles, the latter being represented by specific behaviors (erratic movements and freezing) and avoidance of anxiety-eliciting areas of the test tank (top half and distance from the side walls). We observed that 2D projections of 3D trajectories (ground truth data) may introduce a source of unwanted variation in zebrafish behavioral phenotyping. Predictably, both 2D views underestimate absolute levels of general locomotion. Additionally, while data obtained from a camera positioned on top of the experimental tank are similar to those obtained from a 3D reconstruction, 2D front view data yield false negative findings.
KW - Anxiety
KW - Automated tracking
KW - Citalopram
KW - Ethanol
KW - Novel tank diving test
UR - http://www.scopus.com/inward/record.url?scp=85074302439&partnerID=8YFLogxK
UR - http://www.scopus.com/inward/citedby.url?scp=85074302439&partnerID=8YFLogxK
U2 - 10.7717/peerj.7893
DO - 10.7717/peerj.7893
M3 - Article
C2 - 31637136
AN - SCOPUS:85074302439
SN - 2167-8359
VL - 7
JO - PeerJ
JF - PeerJ
IS - 10
M1 - 7893
ER -