Regional expression patterns of taste receptors and gustducin in the mouse tongue

Kim, Mi-Ryung; Kusakabe, Yuko; Miura, Hirohito; Shindo, Yoichiro; Ninomiya, Yuzo; Hino, Akihiro

doi:10.1016/j.bbrc.2003.10.137

Cited by 141 publications

(113 citation statements)

References 23 publications

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“…The role of the taste-specific Gα protein α-gustducin (29), which is expected to activate a phosphodiesterase, is less clear (28). In taste buds, α-gustducin is inconsistently coexpressed with T1R1-3, particularly in the posterior tongue (30), and its genetic ablation diminishes, but does not abrogate, bitter and umami gustation (31,32). Similarly, the vast majority (≥90%) of cholinergic urethral brush cells respond to bitter (denatonium) and umami (L-glutamate), but only approximately one-third coexpress α-gustducin.…”

Section: Discussionmentioning

confidence: 99%

Bitter triggers acetylcholine release from polymodal urethral chemosensory cells and bladder reflexes

Deckmann

Filipski

Krasteva‐Christ

et al. 2014

Proc. Natl. Acad. Sci. U.S.A.

146

214

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Significance We report the presence of a previously unidentified cholinergic, polymodal chemosensory cell in the mammalian urethra, the potential portal of entry for bacteria and harmful substances into the urogenital system. These cells exhibit structural markers of respiratory chemosensory cells (“brush cells”). They use the classical taste transduction cascade to detect potential hazardous compounds (bitter, umami, uropathogenic bacteria) and release acetylcholine in response. They lie next to sensory nerve fibers that carry acetylcholine receptors, and placing a bitter compound in the urethra enhances activity of the bladder detrusor muscle. Thus, monitoring of urethral content is linked to bladder control via a previously unrecognized cell type.

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Section: Discussionmentioning

confidence: 99%

Bitter triggers acetylcholine release from polymodal urethral chemosensory cells and bladder reflexes

Deckmann

Filipski

Krasteva‐Christ

et al. 2014

Proc. Natl. Acad. Sci. U.S.A.

146

214

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“…Gustducin-expressing cells (mostly bitter taste cells in posterior tongue) generally did not overlap with T1R3-expressing cells (Fig. 3A), reminiscent of their segregation in the circumvallate papilla in posterior tongue (27). Of particular note, these cells displayed morphology typical of mature taste receptor cells, with a characteristic large, round nucleus and a slender cell body (Fig.…”

mentioning

confidence: 95%

Single Lgr5- or Lgr6-expressing taste stem/progenitor cells generate taste bud cells ex vivo

Ren

Lewandowski

Watson

et al. 2014

Proc. Natl. Acad. Sci. U.S.A.

179

189

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Leucine-rich repeat-containing G protein-coupled receptor 5 (Lgr5) and its homologs (e.g., Lgr6) mark adult stem cells in multiple tissues. Recently, we and others have shown that Lgr5 marks adult taste stem/progenitor cells in posterior tongue. However, the regenerative potential of Lgr5-expressing (Lgr5 + ) cells and the identity of adult taste stem/progenitor cells that regenerate taste tissue in anterior tongue remain elusive. In the present work, we describe a culture system in which single isolated Lgr5 + or Lgr6 + cells from taste tissue can generate continuously expanding 3D structures ("organoids"). Many cells within these taste organoids were cycling and positive for proliferative cell markers, cytokeratin K5 and Sox2, and incorporated 5-bromo-2'-deoxyuridine. Importantly, mature taste receptor cells that express gustducin, carbonic anhydrase 4, taste receptor type 1 member 3, nucleoside triphosphate diphosphohydrolase-2, or cytokeratin K8 were present in the taste organoids. Using calcium imaging assays, we found that cells grown out from taste organoids derived from isolated Lgr5 + cells were functional and responded to tastants in a dose-dependent manner. Genetic lineage tracing showed that Lgr6 + cells gave rise to taste bud cells in taste papillae in both anterior and posterior tongue. RT-PCR data demonstrated that Lgr5 and Lgr6 may mark the same subset of taste stem/progenitor cells both anteriorly and posteriorly. Together, our data demonstrate that functional taste cells can be generated ex vivo from single Lgr5 + or Lgr6 + cells, validating the use of this model for the study of taste cell generation. Lgr5 (leucine-rich repeat-containing G protein-coupled receptor 5), encoded by a Wnt (wingless-type MMTV integration site family) target gene, marks adult stem/progenitor cells in taste tissue in posterior tongue that in vivo give rise to all major types of taste bud cells, as well as perigemmal cells (6, 7). Lgr5 is also known to mark actively cycling stem cells in small intestine, colon, stomach, and hair follicle, as well as quiescent stem cells in liver, pancreas, and cochlea (8). Isolated Lgr5 + adult stem cells from multiple tissues are able to generate so-called organoid structures ex vivo (9-11). For instance, Sato and colleagues (10) developed a 3D culture system to grow crypt-villus organoids from single intestinal stem cells; all differentiated cell types were found in these structures, indicating the multipotent nature of these cells. We hypothesized that Lgr5 + taste stem/progenitor cells in a 3D culture system would be capable of expanding and giving rise to taste receptor cells ex vivo. In the present study, we isolated Lgr5 + stem/progenitor cells from taste tissue and cultured them in a 3D culture system. Single Lgr5 + cells grew into organoid structures ex vivo in defined culture conditions, with the presence of both proliferating cells and differentiated mature taste cells in which taste signaling components are functionally expressed. When organoids were replated onto a 2D sur...

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“…16,17) Accordingly, we examined the expression pattern of Fxyd6 in fungiform papillae. Robust signals for Fxyd6 were observed in the fungiform papillae at the tip of the tongue, similarly to expression in the CV (Fig.…”

Section: Resultsmentioning

confidence: 99%

“…CV epithelium samples were obtained as previously described. 16) Other first-strand cDNAs from control tissues (brain, kidney, lung, muscle, heart, and stomach) were purchased from GenoStaff (Tokyo). The cDNAs of FXYD family members and the Na,K-ATPase and subunits were amplified using oligonucleotide primer pairs using previously published sequences (Table 1).…”

Section: Methodsmentioning

confidence: 99%

“…In situ hybridization experiments were performed as previously described. 16) CV and fungiform papillae from 8-week-old male mice were frozen in optimum cutting temperature (OCT) compound (Sakura Finetech USA, Los Angeles, CA), and subsequently sectioned into 5-mm thick slices. All hybridization reactions were performed at 65 C. In the case of single-colored in situ hybridization, signals were detected using an alkaline phosphatase-conjugated anti-digoxigenin antibody (Roche; 1:400), in combination with 5-bromo-4-chloro-3-indolyl-phosphate (BCIP) and …”

Section: Methodsmentioning

confidence: 99%

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FXYD6, a Na,K-ATPase Regulator, Is Expressed in Type II Taste Cells

Shindo

Morishita

Kotake

et al. 2011

Bioscience, Biotechnology, and Biochemistry

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Regional expression patterns of taste receptors and gustducin in the mouse tongue

Cited by 141 publications

References 23 publications

Bitter triggers acetylcholine release from polymodal urethral chemosensory cells and bladder reflexes

Bitter triggers acetylcholine release from polymodal urethral chemosensory cells and bladder reflexes

Single Lgr5- or Lgr6-expressing taste stem/progenitor cells generate taste bud cells ex vivo

FXYD6, a Na,K-ATPase Regulator, Is Expressed in Type II Taste Cells

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