基于单细胞转录组测序技术对鄂尔多斯细毛羊次级毛囊形态发生诱导期的研究

doi:10.16431/j.cnki.1671-7236.2022.11.020

Abstract

Abstract: 【Objective】 The purpose of this experiment was to analyze the molecular characteristics and differentiation process of main cell types in the morphogenesis of the secondary hair follicles in Erdos fine wool sheep during the embryonic stage.【Method】 HE staining was performed on skin samples from the lateral part (posterior edge of scapula) of 87 fetal age of 3 Ordos fine wool sheep to identify the development period of hair follicles.Single-cell transcriptome sequence (scRNA-Seq) was performed on some skin samples after mixing.Cell clusters were analyzed by t-distributed stochastic Neigh or embedding (tSNE), and dermal lineage cells and epidermal lineage cells were identified by Col1a1 and Krt15 genes, respectively.The marker genes of different skin cells were used for cell type analysis.The sequence data were analyzed to explore the expression of differential genes in the process of differentiation.The function of the gene was further verified by GO function enrichment analysis.【Result】 HE staining results showed that Ordos fine wool sheep were in the secondary hair follicle induction stage at 87 days of gestational age. 10 603 cells and 18 704 genes were obtained by scRNA-Seq in skin cell samples from the body side of fine wool sheep at 87 days of gestational age could be used for downstream analysis.Based on tSNE analysis, 15 cell clusters were found in scRNA-Seq data.The identification of Col1a1 and Krt15 marker genes showed that dermal and epidermal cells were highly heterogeneous.The differentiation trajectories and dynamic gene expression profiles of dermal/epidermal cell lineage in the process of hair follicle morphogenesis were constructed by quasi-sequential analysis, during the differentiation of dermal lineate cells from fibroblast (Fb) to mature dermal condensates (DC), several different stage marker genes FST recombinant protein (Fst), inhibin subunit βα(Inhba) and transcriptional repressor GATA binding 1(Trps1) were specifically expressed on the quasisequential track, and Wnt, Noggin, bone morphogenetic protein (BMP) and other signaling pathways related to hair follicle morphogenesis were enriched.During the differentiation of epidermal lineage cells, the marker genes Krt10, HOXC13 and SHH of matrix and interfollicle epidermis (IFE) were specifically expressed in both branches of the quasi-sequential trajectory of epidermal lineage cells, and the pathways related to cell proliferation and cell adhesion were enriched.【Conclusion】 The results showed that dermal lineage cells differentiated from Fb to DC during the induction of the secondary hair follicles in fine wool sheep.Epidermal lineage cells were in the stage of proliferation and differentiation of epidermal cells between matrix and hair follicle.The results of this study could enrich people's understanding of the morphogenesis of the secondary hair follicles in fine wool sheep, and provide strong theoretical and technical support for the breeding of Ordos fine wool sheep.

Key words: Ordos fine wool sheep; secondary hair follicles; single-cell transcriptome sequence(scRNA-Seq); hair follicle development

CLC Number:

S826.9⁺1

HE Xue, GU Ying, SIDENG Danba, DE Dema, DOU Xiaoning, WU Yi, LEI Zhihui, PAN Heping, YUE Yaojing. Induction of Secondary Hair Follicle Morphogenesis in Ordos Fine Wool Sheep Based on Single-cell Transcriptome Sequencing Technology[J]. China Animal Husbandry and Veterinary Medicine, 2022, 49(11): 4296-4306.

References

[1] JAMES K, HOSKING B, GARDNER J, et al.Sox18 mutations in theragged mouse allelesragged-like andopossum[J].Genesis, 2003, 36(1):1-6.
[2] ZHANG Y H, TOMANN P, ANDL T, et al.Reciprocal requirements for EDA/EDAR/NF-kappa B and Wnt/β-catenin signaling pathways in hair follicle induction[J]. Developmental Cell, 2009, 17(1):49-61.
[3] SAXENA N, MOK K, RENDL M.An updated classification of hair follicle morphogenesis[J].Experimental Dermatology, 2019, 228(1):55-61.
[4] GUPTA K, LEVINSOHN J, LINDERMAN G, et al.Single-cell analysis reveals a hair follicle dermal niche molecular differentiation trajectory that begins prior to morphogenesis[J].Developmental Cell, 2018, 48(1):17-31.
[5] 李国强, 纪影畅, 李宇.毛囊形态发生的分子机制[J].国际皮肤性病学杂志, 2004, 30(1):38-40. LI G Q, JI Y C, LI Y, et al.Molecular mechanisms of hair follicle morphogenesis[J].International Journal of Dermatology and Venereology, 2004, 30(1):38-40.(in Chinese)
[6] EZRATTY E, STOKES N, CHAI S, et al.A role for the primary cilium in notch signaling and epidermal differentiation during skin development[J]. Cell, 2011, 145(7):1129-1141.
[7] PAUS R, MüLLER-RVER S, VEEN C, et al.A comprehensive guide for the recognition and classification of distinct stages of hair follicle morphogenesis[J].Journal of Investigative Dermatology, 1999, 113(4):523-532.
[8] SCHMIDT-ULLRICH R, PAUS R.Molecular principles of hair follicle induction and morphogenesis[J].Bioessays, 2005, (3):66-79.
[9] WANG Y, NAVIN N.Advances and applications of single-cell sequencing technologies[J].Molecular Cell, 2015, 58(4):598-609.
[10] PHAN Q M, SINHA S, BIERNASKIE J, et al.Single-cell transcriptomic analysis of small and large wounds reveals the distinct spatial organization of regenerative fibroblasts[J].Experimental Dermatology, 2021, 9(4):79-106.
[11] TAKEO M, LEE W.Wound healing and skin regeneration[J]. Cold Spring Harbor Perspectives in Medicine, 2015, 5(1):a023267.
[12] MORGAN B A.The dermal papilla:An instructive niche for epithelial stem and progenitor cells in development and regeneration of the hair follicle[J].Cold Spring Harbor Perspectives in Medicine, 2014, 4(7):a015180.
[13] YANG H, ADAM R C, GE Y, et al.Epithelial-mesenchymal micro-niches govern stem cell lineage choices[J]. Cell, 2017, 169(3):483-496.
[14] LEGUE E.Hair follicle renewal:Organization of stem cells in the matrix and the role of stereotyped lineages and behaviors[J]. Development, 2005, 132(18):4143.
[15] BIGGS L C, MÄKELÄ O J, MYLLYMÄKI S M, et al.Hair follicle dermal condensation forms via FGF20 primed cell cycle exit, cell motility, and aggregation[J].eLife Sciences, 2018, 7(7):e36468.
[16] GLOVER J D, WELLS K L, FRANZISKA M, et al.Hierarchical patterning modes orchestrate hair follicle morphogenesis[J].PLoS Biology, 2017, 15(7):e2002117.
[17] JUN J I, LAU L F.The matricellular protein CCN1 induces fibroblast senescence and restricts fibrosis in cutaneous wound healing[J].Nature Cell Biology, 2010, 12(7):676.
[18] GE W, TAN S J, WANG S H, et al.Single-cell transcriptome profiling reveals dermal and epithelial cell fate decisions during embryonic hair follicle development[J]. Theranostics, 2020, 10(17):7581-7598.
[19] SENNETT R, WANG Z, REZZA A, et al.An integrated transcriptome atlas of embryonic hair follicle progenitors, their niche, and the developing skin[J].Developmental Cell, 2015, 34(5):577-591.
[20] REZZA A, WANG Z, SENNETT R, et al.Signaling networks among stem cell precursors, transit-amplifying progenitors, and their niche in developing hair follicles[J].Cell Reports, 2016, 14(12):3001-3018.
[21] SCHLAKE T.Determination of hair structure and shape[J].Seminars in Cell & Developmental Biology, 2007, 18(2):267-273.
[22] HUH S H, NARHI K, LINDFORS P H, et al.FGF20 governs formation of primary and secondary dermal condensations in developing hair follicles[J]. Genes & Development, 2013, 27(4):450-458.
[23] TUMBAR T.Hairy tale of signaling in hair follicle development and cycling[J].Seminars in Cell & Developmental Biology, 2012, 23(8):906-916.
[24] RISHIKAYSH P, DEV K, DIAZ D, et al.Signaling involved in hair follicle morphogenesis and development[J].International Journal of Molecular Sciences, 2014, 15(1):1647-1670.
[25] GRECO V, CHEN T, RENDL M, et al.A two-step mechanism for stem cell activation during hair regeneration[J].Cell Stem Cell, 2009, 4(2):155-169.
[26] D HEN, JARRELL A, GUO C, et al.Dermal β-catenin activity in response to epidermal Wnt ligands is required for fibroblast proliferation and hair follicle initiation[J]. Development, 2012, 139(8):1522-1533.
[27] AHTIAINEN L, LEFEBVRE S LINDFORS P, et al.Directional cell migration, but not proliferation, drives hair placode morphogenesis[J]. Developmental Cell, 2014, 28(5):588-602.

Induction of Secondary Hair Follicle Morphogenesis in Ordos Fine Wool Sheep Based on Single-cell Transcriptome Sequencing Technology

PDF (PC)

Knowledge

Abstract

Cite this article

share this article

References

Related Articles 3

Recommended Articles

Metrics

Comments

[1]	QIN Zhiyun, JIANG Huaizhi. An Overview of Research on Skin Hair Follicle Development and Regulatory Mechanism in Sheep During the Fetal Period [J]. China Animal Husbandry and Veterinary Medicine, 2023, 50(2): 638-646.
[2]	WANG Dong, LIANG Jiayue, WANG Xiaobing, FENG Juan, ZHANG Ru, XIA Huiyan, XIAO Hongmei. Research Progress of Non-coding RNA in the Development of Hair Follicles of Cashmere Goats [J]. China Animal Husbandry and Veterinary Medicine, 2022, 49(5): 1817-1827.
[3]	DING Yangyang, ZHAO Huijing, GUAN Weijun, HE Xiaohong, PU Yabin, JIANG Lin, MA Yuehui, ZHAO Qianjun. Isolation and Identification of Goat Dermal Papilla Cells and Expression of TGF/β-smad Pathway Related Genes [J]. China Animal Husbandry and Veterinary Medicine, 2020, 47(9): 2724-2731.