Taste Bud Cell Differentiation from SOX10-Expressing Progenitors in the Connective Tissue

味蕾细胞从结缔组织中表达 SOX10 的祖细胞分化

• 批准号: 9809052
• 负责人: Hongxiang Liu
• 金额: $ 17.75万
• 依托单位: UNIVERSITY OF GEORGIA
• 依托单位国家: 美国
• 财政年份: 2019
• 资助国家: 美国
• 起止时间: 2019-06-12 至 2021-05-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9809052
• 关键词: Abbreviations; Biological Models; Birth; Cell Differentiation process; Cells; Connective Tissue; Connective Tissue Cells; Data; Defect; Derivation procedure; Development; Differentiation Antigens; Dose; Eating; Embryo; Epithelial; Epithelial Cells; Epithelium; Esthesia; Future; Heterogeneity; Homeostasis; In Vitro; Knowledge; LGR5 gene; Label; Life; Location; Longevity; Maintenance; Mammals; Mesenchymal; Mesenchyme; Modeling; Molecular; Mus; Neural Crest; Neural Crest Cell; Organ; Perinatal; Population; Pregnancy; Primordium; Publishing; Quality of life; Regulation; Reporting; Sensory; Shapes; Source; Stem cells; Structure; Structure of gustatory pore; Syncope; System; Tamoxifen; Taste Bud Cell; Taste Buds; Taste Disorders; Taste Perception; Testing; Time; Tissues; Tongue; base; cell type; in vivo; postnatal; progenitor; promoter; three dimensional cell culture; tongue papilla

Project Summary/Abstract (abbreviations: taste bud (TB); connective tissue (CT); neural crest (NC)) In mammals, the sensory end organs for taste are taste buds (TBs), which reside primarily in the epithelium of taste papillae in the tongue. Although the location and shape of taste papillae (fungiform, foliate, and circumvallate) vary, each papilla is composed of an epithelium containing TBs, and an underlying mesenchyme/connective tissue (CT) core including various cell types derived primarily from the neural crest (NC). Like other epithelial cells, TB cells have a short lifespan and undergo continuous turnover. Therefore, progenitor/stem cells in the surrounding tissue compartments must be available to renew taste cells for homeostasis. Information regarding these TB progenitors is necessary to understand both TB formation and maintenance, and how taste disorders emerge from TB progenitor defects. TBs emerge perinatally, and undergo maturation and maintenance postnatally. Our recent findings indicate that a previously unappreciated source of progenitors, SOX10-expressing (hereafter SOX10+) progenitors, may differentiate into a unique population of taste cells. Using a model of Cre under the control of the SOX10 endogenous promoter, which is NC-specific in early embryos, we found that SOX10-Cre labeled a unique population of TB cells (faint for the widely used pan-taste cell marker K8) concurrently with the underlying CT, which originates primarily from the NC. These data suggest that (1) SOX10+ cells in NC-derived CT are potentially progenitors that differentiate to unique TB cells during TB maturation and maintenance, and (2) TB cells are more diverse than previously thought. Further, we identified tamoxifen-induced SOX10-iCreERT2/RFP cells in the CT core of taste papillae in 2-wk-old mice while labeled cells were seen in TBs at 2 wks after tamoxifen. This new data suggests the existence of SOX10+ progenitors in the CT for TB cell renewal, warranting further studies. In this application, we will test our hypothesis that SOX10+ TB progenitors exist in the underlying CT, undergo a transition from mesenchyme to TB-surrounding epithelium, and differentiate to form unique TB cells. (Aim 1) To demonstrate firmly the contribution of SOX10+ progenitors to specific TB cell types, we will administer tamoxifen to inducible SOX10-iCreERT2/RFP mice at different postnatal stages and trace SOX10+ progenitors and labeled TB cells at various time points after tamoxifen treatment. (Aim 2) To determine whether NC cells give rise to SOX10+ progenitors and the derived unique TB cells, we will provide a single dose of tamoxifen to dams at ~E8.5 pregnancy of SOX10-iCreERT2/RFP embryos, and determine whether SOX10-iCreERT2 labels SOX10+ CT cells. We will also determine whether the labeled cells localize within the TBs of postnatal mice. Finally, (Aim 3) we will demonstrate the differentiation of isolated SOX10+ mesenchymal progenitors to TB cells in 3D cultures in vitro. Demonstration of the contribution, derivation, and differentiation of SOX10+ TB progenitors will significantly advance our knowledge regarding the progenitor source(s) required for TB cell renewal, and thereby taste as a whole. Key words: taste bud (TB); connective tissue (CT); neural crest (NC); SOX10; 3D cell cultures.

项目总结/摘要（缩写：味蕾（TB）；结缔组织（CT）；神经嵴（NC）） 在哺乳动物中，味觉末端器官是味蕾 (TB)，主要位于上皮细胞中 舌头上的味觉乳头。尽管味觉乳头（菌状、叶状和 周围) 有所不同，每个乳头都由含有 TB 的上皮和底层组成 间充质/结缔组织 (CT) 核心，包括主要源自神经嵴的各种细胞类型 （NC）。与其他上皮细胞一样，结核细胞的寿命很短，并且会不断更新。所以， 周围组织区室中的祖细胞/干细胞必须可用于更新味觉细胞 体内平衡。有关这些结核病祖细胞的信息对于了解结核病的形成和 维持，以及味觉障碍如何因结核祖细胞缺陷而出现。 结核病在围产期出现，并在产后成熟和维持。我们最近的研究结果表明 以前未被重视的祖细胞来源，表达 SOX10（以下简称 SOX10+）祖细胞，可能 分化成独特的味觉细胞群。使用 SOX10 控制下的 Cre 模型 内源性启动子，在早期胚胎中具有 NC 特异性，我们发现 SOX10-Cre 标记了一个独特的 TB 细胞群（对广泛使用的泛味细胞标记物 K8 感到晕眩）与基础 CT 同时进行， 主要源自 NC。这些数据表明 (1) NC 衍生 CT 中的 SOX10+ 细胞可能是 在 TB 成熟和维持过程中分化为独特 TB 细胞的祖细胞，以及 (2) TB 细胞更 与之前想象的不同。此外，我们在 CT 中鉴定了他莫昔芬诱导的 SOX10-iCreERT2/RFP 细胞 在服用他莫昔芬后 2 周，在 2 周龄小鼠的味觉乳头核心中观察到标记细胞，而在 TB 中观察到标记细胞。这个新数据 表明 CT 中存在 SOX10+ 祖细胞以促进 TB 细胞更新，值得进一步研究。 在此应用中，我们将检验我们的假设，即 SOX10+ TB 祖细胞存在于底层 CT 中，经过 从间充质到结核周围上皮的转变，并分化形成独特的结核细胞。 （目标1） 为了坚定地证明 SOX10+ 祖细胞对特定 TB 细胞类型的贡献，我们将给予 他莫昔芬对不同产后阶段的可诱导 SOX10-iCreERT2/RFP 小鼠的作用并追踪 SOX10+ 祖细胞 并在他莫昔芬治疗后的不同时间点标记结核细胞。 （目标 2）确定 NC 细胞是否 产生 SOX10+ 祖细胞和衍生的独特 TB 细胞，我们将提供单剂量的他莫昔芬 在 ~E8.5 妊娠 SOX10-iCreERT2/RFP 胚胎时检测母鼠，并确定 SOX10-iCreERT2 是否标记 SOX10+ CT 细胞。我们还将确定标记的细胞是否位于出生后小鼠的结核内。 最后，（目标 3）我们将展示分离的 SOX10+ 间充质祖细胞向 TB 细胞的分化 在体外 3D 培养中。 SOX10+ 结核祖细胞的贡献、衍生和分化的证明将显着 增进我们对结核细胞更新所需祖细胞来源的了解，从而从整体上了解味道。 关键词：味蕾（TB）；结缔组织（CT）；神经嵴（NC）； SOX10； 3D 细胞培养。