Single-Cell RNA-Sequencing From Mouse Incisor Reveals Dental Epithelial Cell-Type Specific Genes
Single-Cell RNA-Sequencing From Mouse Incisor Reveals Dental Epithelial Cell-Type Specific Genes
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DOI：
10.3389/fcell.2020.00841
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发表时间：
2020-09-01
影响因子：
5.5
通讯作者：
Fukumoto, Satoshi
中科院分区：
文献类型：
--
作者：
Chiba, Yuta;Saito, Kan;Fukumoto, Satoshi
牙齿上皮干细胞产生四种类型的牙齿上皮细胞：内釉上皮（IEE）、外釉上皮（OEE）、中间层（SI）和星状网状层（SR）。IEE细胞进一步分化为成釉细胞，成釉细胞发挥不同的作用，是釉质形成所必需的。它们通常按其形状分类，尽管它们的转录组和生物学作用尚未完全了解。在这里，我们的目的是使用单细胞RNA测序来阐明牙齿上皮细胞类型的异质性。来自出生后第7天小鼠切牙的6，260个单细胞的无偏聚类将其分为两个成釉细胞群，IEE/OEE，SI/SR和两个间充质细胞群。分泌期成釉细胞表达Amel和Enam，分为Dspp+和Ambn+两种类型。假时间分析显示Dspp+成釉细胞分化为Ambn+成釉细胞。此外，DsppandAmbn可能是成釉细胞的阶段特异性标志物。每个聚类的基因本体分析表明细胞类型的潜在作用：OEE在牙齿大小的调节和SR在营养物质的运输。随后，我们确定了新的牙上皮细胞标记基因，命名为Pttg 1，Atf 3，Cldn 10和Krt 15。这一结果不仅为牙细胞的转录组数据提供了来源，而且有助于釉质形成的分子分析。
Dental epithelial stem cells give rise to four types of dental epithelial cells: inner enamel epithelium (IEE), outer enamel epithelium (OEE), stratum intermedium (SI), and stellate reticulum (SR). IEE cells further differentiate into enamel-forming ameloblasts, which play distinct roles, and are essential for enamel formation. These are conventionally classified by their shape, although their transcriptome and biological roles are yet to be fully understood. Here, we aimed to use single-cell RNA sequencing to clarify the heterogeneity of dental epithelial cell types. Unbiased clustering of 6,260 single cells from incisors of postnatal day 7 mice classified them into two clusters of ameloblast, IEE/OEE, SI/SR, and two mesenchymal populations. Secretory-stage ameloblasts expressedAmelandEnamwere divided intoDspp+ andAmbn+ ameloblasts. Pseudo-time analysis indicatedDspp+ ameloblasts differentiate intoAmbn+ ameloblasts. Further,DsppandAmbncould be stage-specific markers of ameloblasts. Gene ontology analysis of each cluster indicated potent roles of cell types: OEE in the regulation of tooth size and SR in the transport of nutrients. Subsequently, we identified novel dental epithelial cell marker genes, namelyPttg1,Atf3,Cldn10, andKrt15. The results not only provided a resource of transcriptome data in dental cells but also contributed to the molecular analyses of enamel formation.