ZFIN Image: Lescouzères et al., 2023, Figure 2

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Figure Caption

Figure 2 Development of a screening strategy in zebrafish to score, categorize, and refine candidates to 103 hits

Efficacy of Hits is determined by the establishment of a z‐score scale, ranging from 0 (noninjected “WT Control,” green) to negative values < −2 (gan morpholino‐injected “MO Control,” red) after normalization with WT values. Representative pictures of video tracking show the total distance traveled over 1 h by quadruplicate WT and MO fish at 5 dpf.

Primary screening on gan morphants identifies four chemicals classes depending on their mean z‐scores and individual values. Hits restore motility in > 2/4 animals (min 2 individual z‐scores > −1), while non‐Hits may have an effect only on 1/4 animals (max 1 z‐score > −1) and toxic compounds induce death in ≥ 3 fish. Hits are further categorized in two groups A and B, accordingly to the penetrance of the phenotype: B Hits exhibit a negative mean z‐scores due to 1–2 fish with low values, while A Hits are penetrant with positive mean z‐scores. Screening of the 1,280 Prestwick Chemical compounds identifies 809 Toxic and 368 Non‐Hits molecules, and a total of 103 Hits with 71 A Hits and 32 B Hits.

Summary of the screening strategy workflow for the GAN disease, encompassing behavioral tests in the two gan zebrafish models (Screen and Rescreen), computational approach using System Biology (Filter I) and a novel imaging screening methodology at the cellular level in zebrafish (Filters II 1–3).

Acknowledgments

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