Depletion of otoferlin results in reduced hair cell vesicle recycling. (A–A’) Representative confocal images of 96 hpf (A) phalloidin stained neuromasts and (A’) phalloidin and mCLING stained neuromast viewed laterally. (A”) A diagram of a neuromast viewed laterally, with phalloidin stained actin denoted in red, and mCLING loaded hair cells denoted in green. Support cells are depicted in grey. (B–B”) Representative confocal images of control morpholino injected (B) and otoferlin morphant injected (B’) lateral line neuromasts labeled with DAPI (blue) and mCLING (green) of mCLING incubation at room temperature. (B”) A diagram of a neuromast in the same orientation as B, B’, with DAPI denoted in blue, and mCLING loaded hair cells denoted in green. Support cells are depicted in grey. (C). Quantification of average mCLING dye associated with neuromasts of negative control injected and morphant zebrafish (t-test, p < 0.001). N = 6 larvae, 3–6 neuromasts per larvae for both negative control and morphant. (D–D’) Representative images of lateral line neuromasts stained for otoferlin (green) under (D) control injected and (D’) morpholino injected conditions. Scale bars = 5 µm.
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