Proteomic Analysis of Skin Tissue in IGF-I Transgenic Fine Wool Sheep (Ovis aries) Based on DIA Technology
ZHANG Yi-Yuan, GUO Yan-Hua, TANG Hong, GAO Lei, WANG Xin-Hua, ZHOU Ping, WANG Li-Min*
Research Institute of Animal Husbandry and Veterinary Medicine, Xinjiang Academy of Agricultural and Reclamation Sciences/State Key Laboratory for Sheep Genetic Improvement and Healthy Production, Shihezi 832000, China
Abstract：The wool yield of transgenic Chinese Merion fine wool sheep (Ovis aries) with insulin-like growth factor I (IGF-I) gene was significantly increased. In order to find out the change of skin protein expression related to wool production, data independent acquisition (DIA) proteome sequencing technology and bioinformatics analysis were used to analyze the total skin protein of transgenic sheep and wild-type sheep, which aimed to find specific proteins related to wool growth characteristics. Firstly, DIA proteomics method and liquid chromatography tandem mass spectrometry (LC-MS/MS) were used to sequence the tissues of IGF-I transgenic sheep and wild type sheep. The sequencing data were normalized, differentially expressed proteins were analyzed, Gene Ontology (GO) and KEGG enrichment analysis and candidate protein screening were carried out. The results showed that 2 914 proteins were obtained, including 930 differentially expressed proteins. Compared with wild type, 374 proteins were up-regulated and 556 proteins were down-regulated, and 18 differentially expressed proteins were related to wool traits. GO analysis showed that there were 24, 14 and 18 annotations to biological process, molecular function and cell component, respectively; KEGG pathway analysis showed that the differentially expression proteins were involved in 42 signaling pathways, including 8 signaling pathways such as serine/threonine kinase, secretory glycoprotein, gonadotropin releasing hormone, tumor suppressor protein, transforming growth factor-β, which was related to wool growth and development. This study is helpful to reveal the relationship between differential protein and wool traits and provides a reference for exploring the molecular mechanism of wool traits and molecular marker breeding.
[1] 陈甜, 肖海峰. 2014. 中国羊毛质量研究[J]. 农产品质量与安全, (5): 66-71. (Chen T, Xiao H F. 2014. Study on the quality of chinese wool[J]. Quality and Safety of AGRO-Products, (5): 66-71.) [2] 康晓龙. 2013. 基于转录组学滩羊卷曲被毛形成的分子机制研究[D]. 博士学位论文, 中国农业大学, 导师: 吴常信, pp. 53-56. (Kang X L.2013. Transcriptome study on the molecular mechanism of curly coat phenotype in Chinese Tan sheep[D]. Thesis for Ph.D., China Agricultural University, Supervisor: Wu C X, pp.53-56.) [3] 李少斌. 2018. 世界羊毛业现状和发展趋势[J]. 养殖与饲料, (11): 5-8. (Li S B. 2018. Current situation and development trend of world wool industry[J]. Animals Breeding and Feed, (11): 5-8.) [4] 申正. 2017. IGF1和TIMP1对绒山羊毛乳头细胞发育过程中相关信号通路的调控研究[D]. 硕士学位论文, 内蒙古大学, 导师: 刘东军, pp.Ⅰ-Ⅴ. (Shen Z.2017. Regulation of IGF1 and TIMP1 on the related signal pathways during development of arbas cashmere goat dermal papilla cells[D]. Thesis for M.S., Inner mongolia University, Supervisor: Liu D J, pp.Ⅰ-Ⅴ.) [5] 杨佐青, 陶金忠, 李颖康, 等. 2017. 应用iTRAQ技术对滩羊裘皮差异表达蛋白的研究[J]. 中国畜牧兽医, 44(9): 2682-2691. (Yang Z Q, Tao J Z, Li Y K, et al.2017. Study on differentially expressed proteins of Tan sheep skin using iTRAQ technology[J]. China Animal Husbandry and Veterinary Medicine, 44(9): 2682-2691.) [6] Almeida A M, Plowman J E, Harland D P, et al.2014. Influence of feed restriction on the wool proteome: A combined iTRAQ and fiber structural study[J]. Journal of Proteomics, 103: 170-177. [7] Christiano A M.2008. Hair follicle epithelial stem cells get their SOX on[J]. Cell Stem Cell, 3(1): 3-4. [8] Collins C A, Watt F M.2008. Dynamic regulation of retinoic acid-binding proteins in developing, adult and neoplastic skin reveals roles for beta-catenin and Notch signalling[J]. Developmental Biology, 324(1): 55-67. [9] Damak S, Su H, Jay N P, et al.1996. Improved wool production in transgenic sheep expressing insulin-like growth factor 1[J]. Biotechnology (NY), 14(2): 185-188. [10] Gao Y, Wang X, Yan H, et al.2016. Comparative transcriptome analysis of fetal skin reveals key genes related to hair follicle morphogenesis in cashmere goats[J]. PLOS ONE, 11(3): e0151118. [11] Guo T, Han J, Yuan C, et al.2020. Comparative proteomics reveals genetic mechanisms underlying secondary hair follicle development in fine wool sheep during the fetal stage[J]. Journal of Proteomics, 223: 103827. [12] Han W, Yang F, Wu Z, et al.2020. Inner Mongolian Cashmere goat secondary follicle development regulation research based on mRNA-miRNA co-analysis[J]. Scientific Reports, 101(1): 4519. [13] He N, Dong Z, Tai D, et al.2018. The role of Sox9 in maintaining the characteristics and pluripotency of Arbas Cashmere goat hair follicle stem cells[J]. Cytotechnology, 704(4): 1155-1165. [14] Jin M, Zhang J Y, Chu M X, et al.2018. Cashmere growth control in Liaoning cashmere goat by ovarian carcinoma immunoreactive antigen-like protein 2 and decorin genes[J]. Asian-Australasian Journal of Animal Sciences, 315(5): 650-657. [15] Li Y, Zhou G, Zhang R, et al.2018. Comparative proteomic analyses using iTRAQ-labeling provides insights into fiber diversity in sheep and goats[J]. Journal of Proteomics, 172: 82-88. [16] Megdiche S, Mastrangelo S, Ben Hamouda M, et al.2019. A combined multi-cohort approach reveals novel and known genome-wide selection signatures for wool traits in merino and merino-derived sheep breeds[J]. Frontiers in Genetics, 10: 1025. [17] Miao C, Yang Y, Li S, et al.2018. Discrimination and quantification of homologous keratins from goat and sheep with dual protease digestion and PRM assays[J]. Journal of Proteomics, 186: 38-46. [18] Plowman J, Thomas A, Perloiro T, et al.2019. Characterisation of white and black merino wools: A proteomics study[J]. Animal: An International Journal of Animal Bioscience, 13(3): 659-665. [19] Purvis I W, Franklin I R.2005. Major genes and QTL influencing wool production and quality: A review[J]. Genetics, Selection, Evolution, 37(Suppl 1): S97-107. [20] Wisniewski J R, Zougman A, Nagaraj N, et al.2009. Universal sample preparation method for proteome analysis[J]. Nature Methods, 6(5): 359-362. [21] Su R, Fu S, Zhang Y, et al.2015. Comparative genomic approach reveals novel conserved microRNAs in Inner Mongolia cashmere goat skin and longissimus dorsi[J]. Molecular Biology Reports, 42(5): 989-995. [22] Sun B, Boxer L, Ransohoff J, et al.2015. CALML5 is a ZNF750- and TINCR-induced protein that binds stratifin to regulate epidermal differentiation[J]. Genes & Development, 29(21): 2225-2230. [23] Su H Y, Jay N P, Gourley T S, et al.1998. Wool production in transgenic sheep: Results from first-generation adults and second-generation lambs[J]. Animal Biotechnol, 9(2): 135-147. [24] Vidal V, Chaboissier M, Lützkendorf S, et al.2005. Sox9 is essential for outer root sheath differentiation and the formation of the hair stem cell compartment[J]. Current Biology, 15(15): 1340-1351. [25] Wu L, Belyaeva O, Adams M, et al.2019. Mice lacking the epidermal retinol dehydrogenases SDR16C5 and SDR16C6 display accelerated hair growth and enlarged meibomian glands[J]. The Journal of Biological Chemistry, 294(45): 17060-17074. [26] Xu Z, Wang W, Jiang K, et al.2015. Embryonic attenuated Wnt/β-catenin signaling defines niche location and long-term stem cell fate in hair follicle[J]. Elife, 4: e10567. [27] Yu Z, Plowman J, Maclean P,et al.2018. Ovine keratome: Identification, localisation and genomic organisation of keratin and keratin-associated proteins[J]. Animal Genetics, 49(5): 361-370. [28] Zhao J, Qin H, Xin J, et al.2020. Discovery of genes and proteins possibly regulating mean wool fibre diameter using cDNA microarray and proteomic approaches[J]. Scientific Reports, 10(1): 7726.
