Whole transcriptome profiling of taste bud cells.

Whole transcriptome profiling of taste bud cells.
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DOI:
10.1038/s41598-017-07746-z
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发表时间:
2017-08-08
期刊:
影响因子:
4.6
通讯作者:
Margolskee RF
Margolskee RF
中科院分区:
综合性期刊3区
文献类型:
--
作者:
Sukumaran SK;Lewandowski BC;Qin Y;Kotha R;Bachmanov AA;Margolskee RF

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对单细胞RNA-Seq数据的分析可以提供对构成复杂组织的单个细胞类型的特定功能的见解。在这里,我们研究了两个不同亚群的小鼠味觉细胞的基因表达:表达TAS1R3的II型细胞和生理鉴定的III型细胞。我们的RNA-Seq文库符合高质量控制标准,并准确捕获了类型II(例如Tas1r基因、Plcb2、Trpm5)和类型III(例如PKD2L1、NCAM、Snap25)标记基因的差异表达。生物信息学分析表明,调节对刺激反应的基因在II型细胞中上调,而与神经元功能相关的通路在III型细胞中上调。我们还鉴定了与趋化和轴突引导相关的高表达基因和途径,为新的味觉细胞整合到味蕾的潜在机制提供了新的见解。我们通过免疫组织化学证实了编码突触(Cplx2和Pclo)和信号素信号通路(Crmp2,PlexinB1,Fes和Sema4a)组件的选定基因的表达,从而验证了我们的结果。这里描述的方法可以为所有味觉细胞亚群提供一个全面的基因表达图谱,并将特别与味蕾和其他组织中的细胞类型相关,这些细胞类型只能通过生理方法识别。
Analysis of single-cell RNA-Seq data can provide insights into the specific functions of individual cell types that compose complex tissues. Here, we examined gene expression in two distinct subpopulations of mouse taste cells: Tas1r3-expressing type II cells and physiologically identified type III cells. Our RNA-Seq libraries met high quality control standards and accurately captured differential expression of marker genes for type II (e.g. the Tas1r genes, Plcb2, Trpm5) and type III (e.g. Pkd2l1, Ncam, Snap25) taste cells. Bioinformatics analysis showed that genes regulating responses to stimuli were up-regulated in type II cells, while pathways related to neuronal function were up-regulated in type III cells. We also identified highly expressed genes and pathways associated with chemotaxis and axon guidance, providing new insights into the mechanisms underlying integration of new taste cells into the taste bud. We validated our results by immunohistochemically confirming expression of selected genes encoding synaptic (Cplx2 and Pclo) and semaphorin signalling pathway (Crmp2, PlexinB1, Fes and Sema4a) components. The approach described here could provide a comprehensive map of gene expression for all taste cell subpopulations and will be particularly relevant for cell types in taste buds and other tissues that can be identified only by physiological methods.
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