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Fig. 9 Screening for genes involved in hearing and vestibular function by F0 approach. (A) The cartoon figure illustrates zebrafish larval sensory systems, including the inner ear and the neuromast in the lateral line. The inner ear contains the anterior macula (am) with an otolith attached to the ciliary bundles of macular hair cells, responsible for balance, and the posterior macula (pm) with an otolith attached to the ciliary bundles of macular hair cells, responsible for hearing. Additionally, there are three cristae for sensing angular rotation. Functional (mechanotransduction) and viable hair cells in the lateral line neuromast can uptake live nuclear dyes such as Yo-Pro-1. (B) A hearing gene screening pipeline was established. Embryos, both with and without microinjection, were rinsed with embryo water daily until 5 dpf for inner ear phenotyping and Yo-Pro-1 uptake experiments. The remaining larvae were individually transferred to 96-well plates for the AEBR assay at 6 dpf. (C) The WISH method was performed to study mRNA expression patterns for hearing genes. The blue arrowheads indicate the lateral line neuromast, the dashed close circle indicates the inner ear, and the red asterisk indicates the sensory epithelium in the inner ear. Stained embryos at 1 dpf for all genes, at 3 dpf for otol1a and otol1b in the left panel, and at 2 dpf for ptprq were displayed in dorsal view. The rest were shown in lateral view, with the anterior to the left. (D) Representative images of inner ear morphology for each F0 knockout. Scale bar = 0.1 mm. Lateral view with the anterior to the left.