Background: Lingual epithelia in the tongue tip are among the most rapidly regenerating tissues, but the mechanism of cell genesis in this tissue is still unknown. Previous study has suggested the existence of multipl...Background: Lingual epithelia in the tongue tip are among the most rapidly regenerating tissues, but the mechanism of cell genesis in this tissue is still unknown. Previous study has suggested the existence of multiple stem cell pools in lingual epithelia and papillae. Like K14+ and Sox2+ cells, NTPDase2+ cells have characteristics of stem cells.Methods: We employed a system using doxycycline to conditionally ablate NTPDase2+ cells in lingual epithelia and papillae by regulated expression of the diphtheria toxin A(DTA) gene. Transgenic lines, which expressed the rtTA gene in NTPDase2+ cells, were produced by pronuclear injection of zygotes from C57 BL/6 mice using the BAC clone RP23-47 P18. The NTPDase2-rtTA transgenic mice were crossed with the TetO-DTA transgenic animals. The double transgenic mice were treated with doxycycline. Doxycycline(Dox) was diluted in 5% sucrose in water to a final concentration of 0.3-0.5 mg/mL and supplied as drinking water.Results: After 15 days of Dox induction, the expression of NTPDase2, Sox2 and K14 was ablated from lingual epithelia. DTA expression in NTPDase2+cells did not inhibit the turnover of GNAT3+ or PLCb2+ cells in taste buds,nor the expression of S100 b beneath lingual epithelia and papillae. After35 days ablation of NTPDase2+ cells, the basic structure of lingual epithelia and papillae remained intact. However, the ratio of cell to total tissue area was decreased in lingual epithelia and circumvallate(CV) papillae. DTA expression also inhibited the regeneration of filiform papillae on the dorsal surface of the tongue tip.Conclusions: These studies provide important insights into the understanding of dynamic equilibrium among the multiple stem cell populations present in the lingual epithelia and papillae.展开更多
基金Shanghai Public Health Clinical Center,Grant/Award Number:KY-GW-2017-06 and KY-GW-2018-11
文摘Background: Lingual epithelia in the tongue tip are among the most rapidly regenerating tissues, but the mechanism of cell genesis in this tissue is still unknown. Previous study has suggested the existence of multiple stem cell pools in lingual epithelia and papillae. Like K14+ and Sox2+ cells, NTPDase2+ cells have characteristics of stem cells.Methods: We employed a system using doxycycline to conditionally ablate NTPDase2+ cells in lingual epithelia and papillae by regulated expression of the diphtheria toxin A(DTA) gene. Transgenic lines, which expressed the rtTA gene in NTPDase2+ cells, were produced by pronuclear injection of zygotes from C57 BL/6 mice using the BAC clone RP23-47 P18. The NTPDase2-rtTA transgenic mice were crossed with the TetO-DTA transgenic animals. The double transgenic mice were treated with doxycycline. Doxycycline(Dox) was diluted in 5% sucrose in water to a final concentration of 0.3-0.5 mg/mL and supplied as drinking water.Results: After 15 days of Dox induction, the expression of NTPDase2, Sox2 and K14 was ablated from lingual epithelia. DTA expression in NTPDase2+cells did not inhibit the turnover of GNAT3+ or PLCb2+ cells in taste buds,nor the expression of S100 b beneath lingual epithelia and papillae. After35 days ablation of NTPDase2+ cells, the basic structure of lingual epithelia and papillae remained intact. However, the ratio of cell to total tissue area was decreased in lingual epithelia and circumvallate(CV) papillae. DTA expression also inhibited the regeneration of filiform papillae on the dorsal surface of the tongue tip.Conclusions: These studies provide important insights into the understanding of dynamic equilibrium among the multiple stem cell populations present in the lingual epithelia and papillae.
