Retinoic Acid Inhibition Alters Intestinal Composition in Zebrafish: a non-genetic model to study Hirschsprung Disease

In this study, zebrafish larvae were treated with diethylaminobenzaldehyde (DEAB), an inhibitor of RA synthesis, to induce phenotypes resembling total colonic aganglionosis. Single-cell RNA sequencing was performed on the intestinal tissue of DEAB-treated zebrafish. Overall design: Tg(phox2bb:GFP) zebrafish embryos were treated with 20 µM DEAB from 24 hpf until 5 dpf. At 5 dpf, zebrafish larvae were euthanized using tricaine methanesulfonate (MS-222) and the intestines were carefully dissected under a stereomicroscope. Dissected intestines were enzymatically dissociated into a single-cell suspension for single-cell RNA sequencing.