Epitope Tagging with Genome Editing in Mice Reveals That the Proton Channel OTOP1 Is Apically Localized and Not...

The gustatory system allows animals to assess the nutritive value and safety of foods prior to ingestion. The first step in gustation is the interaction of taste stimuli with one or more specific sensory receptors that are generally believed to be present on the apical surface of the taste receptor cells. However, this assertion is rarely tested. We recently identified OTOP1 as a proton channel and showed that it is required for taste response to acids (sour) and ammonium. Here, we examined the cellular and subcellular localization of OTOP1 by tagging the endogenous OTOP1 protein with an N-terminal HA epitope (HA-OTOP1). Using both male and female HA-OTOP1 mice and high-resolution imaging, we show that OTOP1 is strictly localized to the apical tips of taste cells throughout the tongue and oral cavity. Interestingly, immunoreactivity is observed in the actin-rich taste pore above the tight junctions defined by zonula occludens-1 (ZO-1) and also immediately below these junctions. Surprisingly, OTOP1 immunoreactivity is not restricted to Type III taste receptor cells (TRCs) that mediate sour taste but is also observed in glia-like Type I TRCs proposed to perform housekeeping functions, a result that is corroborated by scRNA-seq data. The apical localization of OTOP1 supports the contention that OTOP1 functions as a taste receptor and suggests that OTOP1 may be accessible to orally available compounds that could act as taste modifiers.