Small molecule screen in embryonic zebrafish using modular variations to target segmentation

December 1, 2017

... tribute volumes of small molecules into master plates. All working concentrations of 2, 10, and 50 µM, were further diluted from one master plate per experiment for higher accuracy using the Tecan Freedom Evo (Tecan) with a 96-head and directly applied to embryos at 10 hpf at a final volume of 100 µl per well to start the treatment. Propylthiouracil was added at a final concentration of 0. ... to distribute volumes of small molecules into master plates. All working concentrations of 2, 10, and 50 µM, were further diluted from one master plate per experiment for higher accuracy using the Tecan Freedom Evo (Tecan) with a 96-head and directly applied to embryos at 10 hpf at a final volume of 100 µl per well to start the treatment. Propylthiouracil was added at a final concentration

Small molecule in vivo phenotypic screening is used to identify drugs or biological activities by directly assessing effects in intact organisms. However, current screening designs may not exploit the full potential of chemical libraries due to false negatives. Here, we demonstrate a modular small molecule screen in embryonic zebrafish that varies concentration, genotype and timing to target segmentation disorders, birth defects that affect the spinal column. By testing each small molecule in multiple interrelated ways, this screen recovers compounds that a standard screening design would have missed, increasing the hit frequency from the chemical library three-fold. We identify molecular pathways and segmentation phenotypes, which we share in an open-access annotated database. These hits provide insight into human vertebral segmentation disorders and myopathies. This modular screening strategy is applicable to other developmental questions and disease models, highlighting the power of relatively small chemical libraries to accelerate gene discovery and disease study.

Richter, S; Schulze, U; Tomançak, P; Oates, AC;

Journal: Nat Commun Pages: 1901

Original article (29196645)