Abstract
RNA-Seq is a powerful tool in transcriptomic profiling of cells and tissues. We recently identified many more taste buds than previously appreciated in chickens using molecular markers to stain oral epithelial sheets of the palate, base of oral cavity, and posterior tongue. In this study, RNA-Seq was performed to understand the transcriptomic architecture of chicken gustatory tissues. Interestingly, taste sensation related genes and many more differentially expressed genes (DEGs) were found between the epithelium and mesenchyme in the base of oral cavity as compared to the palate and posterior tongue. Further RNA-Seq using specifically defined tissues of the base of oral cavity demonstrated that DEGs between gustatory (GE) and non-gustatory epithelium (NGE), and between GE and the underlying mesenchyme (GM) were enriched in multiple GO terms and KEGG pathways, including many biological processes. Well-known genes for taste sensation were highly expressed in the GE. Moreover, genes of signaling components important in organogenesis (Wnt, TGFβ/ BMP, FGF, Notch, SHH, Erbb) were differentially expressed between GE and GM. Combined with other features of chicken taste buds, e.g., uniquely patterned array and short turnover cycle, our data suggest that chicken gustatory tissue provides an ideal system for multidisciplinary studies, including organogenesis and regenerative medicine.
Highlights
RNA sequencing (RNA-Seq) technology has emerged as a powerful and revolutionary approach to quantify gene expression levels and survey detailed transcriptomic profiling at unprecedented resolution and sensitivity[1, 2]
The palate, the base of the oral cavity and the posterior region of the tongue were dissected in sterile phosphate buffered saline (PBS), and epithelial sheets were separated from underlying connective tissue
We demonstrated that (1) more differentially expressed genes (DEGs) were found between the epithelium (1783 highly expressed) and connective tissue (1906 highly expressed) in the base of the oral cavity (Supplemental Table S4) than palate or posterior tongue (1021 highly expressed in the epithelium and 1730 in the connective tissue of palate; 1512, 1880 in the posterior tongue, listed in Supplemental Tables S5 and S6, respectively); (2) a large overlap of DEGs (640 and 889) between epithelium and connective tissues of the palate, base of oral cavity, and posterior tongue were observed using DESeq[2] and Cuffdiff (Fig. 1)
Summary
RNA sequencing (RNA-Seq) technology has emerged as a powerful and revolutionary approach to quantify gene expression levels and survey detailed transcriptomic profiling at unprecedented resolution and sensitivity[1, 2]. Our recent studies using molecular markers to label chicken taste buds in oral epithelial sheets, i.e., palate, base of the oral cavity and posterior region of the tongue, demonstrated that chicken taste buds, like those of mammals, are distributed in a unique pattern[28]. The clustered taste bud patterning in the oral cavity of chickens is reminiscent of the mammalian soft palate[29] Taken together, these data suggest that taste sensory organs in chickens can potentially provide a system for organogenesis studies, including pattern formation. To better understand the transcriptomic architecture of gustatory tissue in the oral cavity of chickens, RNA-Seq analysis was carried out with the following specific objectives: i) to demonstrate the validity of chicken taste organs as an ideal system for organogenesis studies, ii) to provide new insights into the underlying www.nature.com/scientificreports/. Such information will facilitate studies on mechanisms underlying chicken taste bud formation which will be beneficial for understanding taste organ development in birds and potentially mammals, including humans
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