Abstract

Taste buds are clusters of polarized sensory cells embedded in stratified oral epithelium. In adult mammals, taste buds turn over continuously and are replenished through the birth of new cells in the basal layer of the surrounding non-sensory epithelium. The half-life of cells in mammalian taste buds has been estimated as 8–12 days on average. Yet, earlier studies did not address whether the now well-defined functional taste bud cell types all exhibit the same lifetime. We employed a recently developed thymidine analog, 5-ethynil-2′-deoxyuridine (EdU) to re-evaluate the incorporation of newly born cells into circumvallate taste buds of adult mice. By combining EdU-labeling with immunostaining for selected markers, we tracked the differentiation and lifespan of the constituent cell types of taste buds. EdU was primarily incorporated into basal extragemmal cells, the principal source for replenishing taste bud cells. Undifferentiated EdU-labeled cells began migrating into circumvallate taste buds within 1 day of their birth. Type II (Receptor) taste cells began to differentiate from EdU-labeled precursors beginning 2 days after birth and then were eliminated with a half-life of 8 days. Type III (Presynaptic) taste cells began differentiating after a delay of 3 days after EdU-labeling, and they survived much longer, with a half-life of 22 days. We also scored taste bud cells that belong to neither Type II nor Type III, a heterogeneous group that includes mostly Type I cells, and also undifferentiated or immature cells. A non-linear decay fit described these cells as two sub-populations with half-lives of 8 and 24 days respectively. Our data suggest that many post-mitotic cells may remain quiescent within taste buds before differentiating into mature taste cells. A small number of slow-cycling cells may also exist within the perimeter of the taste bud. Based on their incidence, we hypothesize that these may be progenitors for Type III cells.

Highlights

  • Taste buds are aggregates of 50–100 specialized sensory cells embedded in the stratified oral epithelium

  • Large numbers of EdU+ cells were seen in the basal layer of epithelium within 1 day of the EdU injection (Figure 2A)

  • Over the two days, there was a nearly 3-fold increase in the number of EdU+ epithelial nuclei outside taste buds, relative to initial labeling (Figure 2F), suggesting that on average, initially labeled basal epithelial cells produce progeny through 2 successive asymmetric mitoses

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Summary

Introduction

Taste buds are aggregates of 50–100 specialized sensory cells embedded in the stratified oral epithelium. Cells in taste buds are specialized; each cell detects at most, a subset of compounds that are structurally related or generate a common sensory submodality (e.g. sweet). In keeping with these specializations, the three currently recognized types of taste bud cells exhibit very distinct morphological features, transcriptomes and cellular functions. Type I cells are termed ‘‘glial-like’’ because they appear to function in clearing neurotransmitters [3], ensheath other taste bud cells with lamellar processes [4] and may regulate the ionic milieu [4,5]. Type II (Receptor) cells express G-proteincoupled receptors (GPCR) selective for sweet, bitter or umami tastants and downstream effectors that mediate inositide-mediated

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