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2017 ; 7
(1
): 1962
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Three-dimensional scoring of zebrafish behavior unveils biological phenomena
hidden by two-dimensional analyses
#MMPMID28512334
Macrì S
; Neri D
; Ruberto T
; Mwaffo V
; Butail S
; Porfiri M
Sci Rep
2017[May]; 7
(1
): 1962
PMID28512334
show ga
The study of zebrafish behavior represents a cornerstone upon which basic
researchers promise to advance knowledge in life sciences. Although zebrafish
swim in a three-dimensional (3D) space, their behavior in the lab is almost
exclusively scored in two dimensions, whereby zebrafish are recorded using a
single camera providing 2D videos. Whether this dimensional reduction preserves
the reliability of data has not been addressed. Here we show that, compared to a
3D observation, 2D data are flawed by over-reporting and under-reporting of
locomotory differences. Specifically, we first reconstructed 3D trajectories
through the integration of synchronous information derived from two cameras, and
then compared them with the original 2D views in classical experimental paradigms
assessing shoaling tendency, fear, anxiety, and general locomotion. Our results
suggest that traditional behavioral scoring of individual zebrafish performed in
2D may undermine data integrity, thereby requiring a general reconsideration of
scoring zebrafish behavior to incorporate a 3D approach. We then demonstrate
that, compared to 2D, a 3D approach requires a reduced number of subjects to
achieve the same degree of validity. We anticipate these findings to largely
benefit animal welfare by reducing the number of experimental subjects, without
affecting statistical power.