Subcellular localization of harmonin in mechanosensory hair cells. Co-labeling of wild-type 5 dpf anterior macula hair cells (A–D) shows anti-zebrafish harmonin localization (red) in the hair bundles (green), kinocilia (arrowheads), cell bodies and at synaptic boutons (arrow in C and D). Incubation conditions favoring hair bundle visualization (E–P) show strong colocalization of harmonin within the bundle structure as well as in the cuticular plate in 5 dpf wild types (E–G). Kinocilia labeling can be seen above the bundle region (asterisks). Enhanced signal detection in mutant hair cells (H–J) shows localization persisting in the kinocilia and in existing bundle structures. Kinocilia are more visible in these panels owing to the reduced bundle architecture in the mutant. Protein distribution seems globular and disorganized in the cell bodies (arrowheads). Hair cells of ush1c MO larvae (K–M) show strong knockdown of harmonin, with localization persisting only in kinocilia at this stage, and no signal observed within hair bundles. Injecting the morpholino into the ush1c-/- mutants reduces subcellular localization of harmonin to undetectable levels in hair cells (N–P). Scale bars: 10 μm.
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