Screening Zebrafish Embryos for Bioactive Compounds

Tübingen strain fish were used for the studies and staged as described (64 (link)). For screening, WT zebrafish embryos were arrayed in 96-well plates at 2 h post fertilization. At 4 hpf the embryo media were replaced with 1:100 dilutions of drugs from the ICCB Known Bioactives Library (Enzo Life Sciences) (18 ). For treatments with 50 μM SC-560, 50 μM NS-398, 30 μM indomethacin, or 100 μM dmPGE₂ (American Bioanalytical, Enzo Life Sciences, Santa Cruz), embryos were arrayed in 12-well plates and treated protected from light at 60% epiboly. Drugs were removed at the 28 ss and embryos were fixed in 4% paraformaldehyde.

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Marra A.N., Adeeb B.D., Chambers B.E., Drummond B.E., Ulrich M., Addiego A., Springer M., Poureetezadi S.J., Chambers J.M., Ronshaugen M, & Wingert R.A. (2019). Prostaglandin signaling regulates renal multiciliated cell specification and maturation. Proceedings of the National Academy of Sciences of the United States of America, 116(17), 8409-8418.

Publication 2019

ICCB Known Bioactives Library

Dilutions Dmpge2 Drugs Embryos Fertilization Fish Indomethacin Library Light Ns 398 Paraformaldehyde Sc 560 Strain Zebrafish

Corresponding Organization :

Other organizations : University of Notre Dame, University of Manchester

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Variable analysis

independent variables

Drug treatments from the ICCB Known Bioactives Library (Enzo Life Sciences)
50 μM SC-560
50 μM NS-398
30 μM indomethacin
100 μM dmPGE2 (American Bioanalytical, Enzo Life Sciences, Santa Cruz)

dependent variables

Phenotypic outcomes of the zebrafish embryos

control variables

Tübingen strain zebrafish embryos
Embryo staging as described in the reference (64)
Embryo media replaced at 4 hpf
Embryos arrayed in 12-well plates and treated at 60% epiboly
Drugs removed at the 28 ss
Embryos fixed in 4% paraformaldehyde

controls

Wild-type (WT) zebrafish embryos used for screening

Annotations

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