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.

项目摘要/摘要（缩略语：味蕾（TB）;结缔组织（CT）;神经嵴（NC）） 在哺乳动物中，味觉的感觉末端器官是味蕾（TB），其主要存在于上皮中 舌头上的味觉乳头。虽然味觉乳头的位置和形状（菌状，叶状， 轮廓）各不相同，每个乳头都由含有TB的上皮和下面的 间充质/结缔组织（CT）核心，包括主要来源于神经嵴的各种细胞类型 （NC）中指定。像其他上皮细胞一样，TB细胞的寿命很短，并且会持续更新。因此，我们认为， 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， 体内平衡关于这些TB祖细胞的信息对于理解TB形成和 以及味觉障碍是如何从TB祖细胞缺陷中出现的。 TB在围产期出现，并在出生后成熟和维持。我们最近的发现表明 一种以前未被认识到的祖细胞来源，即表达SOX 10的祖细胞（下文称为SOX 10+），可能 分化成独特的味觉细胞群。使用SOX 10控制下的Cre模型 内源性启动子，这是NC特异性的早期胚胎，我们发现SOX 10-Cre标记的独特的 TB细胞群（对于广泛使用的泛味细胞标志物K8来说是微弱的）与基础CT同时发生， 主要来源于NC。这些数据表明：（1）NC衍生CT中的SOX 10+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细胞更多 比以前想象的要多。此外，我们在CT中鉴定了他莫昔芬诱导的SOX 10-iCreERT 2/RFP细胞， 在2周龄小鼠的味觉乳头的核心，而标记的细胞在他莫昔芬后2周在TB中观察到。这些新数据 提示在CT中存在SOX 10+祖细胞用于TB细胞更新，从而阻止了进一步的研究。 在本申请中，我们将检验我们的假设，即SOX 10 + TB祖细胞存在于基础CT中，经历了 从间充质到TB周围上皮的过渡，并分化形成独特的TB细胞。(Aim第一章 为了坚定地证明SOX 10+祖细胞对特定TB细胞类型的贡献，我们将给予 他莫昔芬对不同出生后阶段的诱导型SOX 10-iCreERT 2/RFP小鼠和痕量SOX 10+祖细胞的影响 并在他莫昔芬处理后的不同时间点标记TB细胞。(Aim 2）为了确定NC细胞是否 产生SOX 10+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， 在约E8.5妊娠的母鼠中，使用SOX 10-iCreERT 2/RFP胚胎，并确定SOX 10-iCreERT 2是否标记 SOX 10 + CT细胞。我们还将确定标记的细胞是否定位在出生后小鼠的TB内。 最后，（目的3）我们将证明分离的SOX 10+间充质祖细胞向TB细胞的分化 in 3D cultures文化in vitro体外. SOX 10 + TB祖细胞的贡献、衍生和分化的证明将显著地 推进我们关于TB细胞更新所需的祖细胞来源的知识，从而作为一个整体品尝。 关键词：味蕾（TB）;结缔组织（CT）;神经嵴（NC）; SOX 10;三维细胞培养。