Maintenance of taste tissues requires continuous replacement of senescent taste cells with new ones generated from adult taste stem cells. This process depends on taste bud innervation, first noted more than a century ago. Until recently, the molecular mechanism underlying this process remained unclear. Prompted by our observation that single Lgr5+ taste stem/progenitor cells can give rise to all different types of mature taste cells in an ex vivo culture system (“taste organoids”) in the absence of nerve input, which suggests that one (or several) of the components in the defined culture medium may be the principal gustatory neuron-produced factor, we set out to identify such factor. We focused on R-spondin, the ligand for the Lgr5 receptor. Our in situ hybridization study showed that R-spondin 2 is expressed in gustatory neurons. Using a gustatory nerve transection model and adenovirus-encoded R-spondin, we showed that exogenous R-spondin can promote taste cell generation despite denervation. Using the organoid culture system, we further showed that R-spondin is required for taste cell generation. R-spondin interacts not only with Lgr5 or its analogs Lgr4/6 but also with two E3 ligases Znrf3/Rnf43 to regulate Wnt signaling (Lgr4/5/6 – positive regulators, Znrf3/Rnf43 - negative regulators). Therefore, we hypothesized that Znrf3/Rnf43 may serve as a brake, controlled by gustatory neuron-produced R-spondin, for maintaining taste tissue homeostasis. Consistent with this model, taste cell hyperplasia occurred in mice lacking Znrf3/Rnf43 in taste stem/progenitor cells, mirroring the effect of exogenous R-Spondin. We further demonstrated that ablating Znrf3/Rnf43 renders neuronal regulation of taste tissue homeostasis dispensable: regeneration of taste cells occurred in the double knockout mice even in the denervation model. In summary, the ternary interaction of R-spondin, Lgr4/5/6, and Znrf3/Rnf43 plays a key role in neuronal regulation of taste stem cells.