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Single-Cell Transcriptomics Reveals the Differentiation and Spatial Signatures underlying Sheep Hair Follicle Heterogeneity and Wool Curvature

ABSTRACT: Here, Single-cell suspensions from the wavy wool and straight wool lambskins were prepared for unbiased single-cell RNA sequencing (scRNA-seq). Based on UAMP analysis, we identified 19 distinct populations from 15,830 single-cell transcriptomes and delineated their cellular identity from specific gene expression profiles. Furtherly, novel marker gene was applied in identifying dermal papilla cells isolated in vitro. By using pseudotime ordering analysis, we successfully constructed the epithelium cell lineage differentiation trajectory and revealed the dynamic gene expression profiles of matrix progenitors’ commitment to the hair shaft and inner root sheath (IRS) cells. Meanwhile, intercellular communication between different cell populations was inferred based on CellChat and the priori knowledge of ligand-receptor pairs, as a result strong intercellular communication and associated signaling pathways were revealed. Besides, to clarify the molecular mechanism of wool curvature, differentially expressed genes in specific cells between straight wool and curly wool were identified and analyzed. Our findings here provide unbiased and systematic view of transcriptional organization of sheep hair follicle, reveal the differentiation and spatial signatures underlying sheep hair follicle heterogeneity and wool curvature, which will provide a valuable resource for understanding the molecular pathways involved in sheep hair follicle development.

ORGANISM(S): Ovis aries

PROVIDER: GSE186204 | GEO | 2022/01/12

REPOSITORIES: GEO

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Project description:Sheep provide considerable materials for the animal fibre industry. Identifying genes of major effect for wool growth would offer strategies for improving the quality and increasing the yield of fine wool. In this study, we employed Agilent Sheep Gene Expression Microarray and proteomic technology to investigate the gene expression patterns of body side skin (more wool growing) in Aohan fine wool sheep (a Chinese indigenous breed) in comparison with groin skin (no wool growing) at the anagen stage of wool follicle. Microarray study revealed that 4772 probes were differentially expressed, including 2071 upregulated and 2701 downregulated probes in the comparisons of body side skin versus groin skin (S/G). The microarray results were verified by means of quantitative PCR. 1099 probes were assigned to unique genes/transcripts. The number of distinct genes/transcripts (annotated) was 926, of which 352 were up-regulated and 574 were down-regulated. In S/G, 13 genes were up-regulated by more than 10-fold, while 60 genes were down-regulated by more than 10-fold. Further analysis revealed that the majority of the genes possibly related to the wool growth could be assigned into the categories including regulation of cell division, intermediate filament, cytoskeletal part and growth factor activity. Several potential gene families might participate in hair growth regulation, including fibroblast growth factors, transforming growth factor-Î², WNTs, insulin-like growth factor, vascular endothelial growth factors and so on. Proteomic analysis also got 196 differentially expressed protein points, of which 121 were identified as single protein points. Furthermore, according to the results at both mRNA and protein levels, similar regulation mechanism of gene activity might be engaged during skin development and embryo development. One ram and two ewes of 12-month-old Aohan fine wool sheep were used in the microarray study. These animals were half sibs (sharing the same father). In August 2010, two areas of full-thickness skin were sampled from the same animal under local anaesthesia: body side skin (wool bearing) and groin skin (non-wool bearing) for microarray and proteomic experiments. The area of each sample was about 1 cm2. All samples were immediately put into collection tubes and stored in liquid nitrogen for RNA and protein extraction. A total of 15, 208 probes were spotted on this Agilent Sheep Gene Expression Microarray (Santa Clara, CA, USA).

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Single-Cell Transcriptomics Reveals the Differentiation and Spatial Signatures underlying Sheep Hair Follicle Heterogeneity and Wool Curvature

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