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Expression Analysis of Melanogenesis Related Genes TYR, TYRP1 and DCT in Tobiano Mongolian Horse (Equus caballus) Skin Tissue |
ZHAO Ruo-Yang1, LI Chao1, BAO Tuge-Qin1, MONGKE Togtokh1, Laxima1, WANG Wen-Xing2, HUANG Bo-Guang2, BAI Dong-Yi1,*, MANG Lai1,* |
1 College of Animal Science, Inner Mongolia Key Laboratory of Equine Genetics/Breeding and Reproduction/Equine Research Center, Scientific Observing and Experimental Station of Equine Genetics/Breeding and Reproduction, Ministry of Agriculture and Rural Affairs, Inner Mongolia Agricultural University, Hohhot 010018, China; 2 Inner Mongolia Zhong Yun Horse Industry Group, Xilinhot 026000, China |
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Abstract Mammals coat color molecular mechanism has always been hotspot in animal genetics and breeding, variety of coat colors in nature arouse the interest of many researchers; horses (Equus caballus) with different complex colors, which is indispensable to the horse registration in breeding, and the molecular mechanisms of horse coat color is very important. Recent years, most researches are focused on breeds, few researches are from the perspective of the individuals and hair follicle pigment synthesis. Since Tobiano Mongolian Horse has unique coat color, the formation mechanism must be more complex. Pigment synthesis involves a large number of genes and complex regulatory networks. Tyrosinase (TYR), tyrosinase-related protein 1 (TYRP1), and tyrosinase-related protein 2 (TYRP2) which is now called dopachrome tautomerase (DCT) are 3 major genes at downstream of melanogenesis pathway. Paraffin-embedded tissue sections were stained with hematoxylin-eosin (HE) to analyze hair follicle structure and pigment distribution. qRT-PCR was used to detect the mRNA expression levels of TYR, TYRP1 and DCT genes in different color skins. The results showed Western blot (WB) and immunofluorescence (IF) were used to quantitatively locate their corresponding proteins. The pigment distribution in different color (black and white) skin tissues and the length of hair follicle dermal papillae was significantly different, meanwhile, the thickness of dermis was different, and the diameter of hair follicle hairball was slightly different. The mRNA and protein expressions of 3 genes were significantly different in different color skin tissues, and the protein expression was basically located in the hair matrix of hair follicle dermal papillae. The results showed TYR, TYRP1 and DCT, downstream genes of melanogenesis, might affect the synthesis of pigment in the different color skin tissues of Tobiano Mongolian horses, and can be the candidate regulatory factors in the process of hair pigment deposition. It will present information for the in-depth study of the molecular mechanism of Tobiano Mongolian Horse coat color and provide reference for other mammals.
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Received: 03 February 2019
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Corresponding Authors:
baidongyi1983@163.com
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[1] 赫晓燕, 郝欢庆, 刘丹丹, 等. 2010. 青年羊驼耳部和背部皮肤组织中miRNA差异表达研究[J]. 中国生物化学与分子生物学报, 26(11): 1016-1022. (He X Y, He H Q, Liu D D, et al.2010. Difference of microRNA expression in the ear and back skin of young alpaca (Lamaacos)[J]. Chinese Journal of Biochemistry and Molecular Biology, 26(11): 1016-1022.) [2] 李丽莎, 李兰会, 李祥龙, 等 . 2017. 水貂PMEL基因启动子活性及转录调控元件分析[J]. 农业生物技术学报, 25(6): 911-920. (Li L S, Li L H, Li X L, et al.2017. Analysis of the promoter activity and transcriptional regulatory elements of PMEL gene in mink (Mustla vison)[J]. Journal of Agricultural Biotechnology, 25(6): 911-920.) [3] 姬改革, 束婧婷, 单艳菊, 等. 2018. 基于表达谱芯片筛选鸡不同部位皮肤组织差异表达基因[J]. 畜牧兽医学报,49(1): 36-45. (Ji G G, Shu J T, Shan Y J, et al.2018. Identification of differentially expressed genes between different positions of chicken skin based on gene expression microarray[J]. Acta Veterinaria et Zootechnica Sinica, 49(1): 36-45.) [4] Botchkareva N V, Khlgatian M, Longley B J, et al.2001. SCF/c-kit signaling is required for cyclic regeneration of the hair pigmentation unit[J]. FASEB Journal: Official Publication of the Federation of American Societies for Experimental Biology, 15(3): 645-658. [5] Brooks S A, Lear T L, Adelson D L, et al.2007. A chromosome inversion near the KIT gene and the Tobiano spotting pattern in horses[J]. Cytogenetic and Genome Research, 119(3-4): 225-230. [6] Brooks S A, Terry R B, Bailey E.2002. A PCR-RFLP for KIT associated with tobiano spotting pattern in horses[J]. Animal genetics, 33(4): 301-303. [7] Chen T, Zhao B, Liu Y, et al.2018. MITF-M regulates melanogenesis in mouse melanocytes[J]. Journal of Dermatological Science, 90(3): 253-262. [8] Cho M, Ryu M, Jeong Y, et al.2009. Cardamonin suppresses melanogenesis by inhibition of Wnt/beta-catenin signaling[J]. Biochemical and Biophysical Research Communications, 390(3): 500-505. [9] Costin G E, Hearing V J.2007. Human skin pigmentation: melanocytes modulate skin color in response to stress[J]. FASEB Journal: Official Publication of the Federation of American Societies for Experimental Biology, 21(4): 976-994. [10] Dietrich J, Menzi F, Ammann P, et al.2015. A breeding experiment confirms the dominant mode of inheritance of the brown coat colour associated with the (496) Asp TYRP1 allele in goats[J]. Animal genetics, 46(5): 587-588. [11] Duffield D A,Goldie P L.1998. Tobiano spotting pattern in horses: Linkage of to with AlA and linkage disequilibrium[J]. The Journal of Heredity, 89(1): 104-106. [12] Haase B, Jude R, Brooks S A, et al.2008. An equine chromosome 3 inversion is associated with the tobiano spotting pattern in German horse breeds[J]. Animal Genetics, 39(3): 306-309. [13] Hrckova Turnova E, Majchrakova Z, Bielikova M, et al.2017. A novel mutation in the TYRP1 gene associated with brown coat colour in the Australian Shepherd dog breed[J]. Animal Genetics, 48(5): 626. [14] Kim N Y, Bhuiyan M S A, Chae H S, et al.2017. Genome-wide association study for tobiano spotting coat color in Korean Jeju x Thoroughbred horse population[J]. Animal Genetics, 48(6): 728-729. [15] Lei M, Guo H, Qiu W, et al.2014. Modulating hair follicle size with Wnt10b/DKK1 during hair regeneration[J]. Experimental Dermatology, 23(6): 407-413. [16] Ludwig A, Pruvost M, Reissmann M, et al.2009. Coat color variation at the beginning of horse domestication[J]. Science (New York, N.Y.), 324(5926): 485. [17] Makpol S, Jam F A, Rahim N A, et al.2014. Comparable down-regulation of TYR, TYRP1 and TYRP2 genes and inhibition of melanogenesis by tyrostat, tocotrienol-rich fraction and tocopherol in human skin melanocytes improves skin pigmentation[J]. La Clinica Terapeutica, 165(1): e39-e45. [18] Moyo D, Gomes M, Erlwanger K H.2018. Comparison of the histology of the skin of the Windsnyer, Kolbroek and Large White pigs[J]. Journal of the South African Veterinary Association, 89(0): e1-e10. [19] Muller-Rover S, Handjiski B, van der Veen C, et al.2001. A comprehensive guide for the accurate classification of murine hair follicles in distinct hair cycle stages[J]. The Journal of Investigative Dermatology, 117(1): 3-15. [20] Nishimura E K.2011. Melanocyte stem cells: A melanocyte reservoir in hair follicles for hair and skin pigmentation[J]. Pigment Cell & Melanoma Research, 24(3): 401-410. [21] Ogawa H, Hattori M.1983. Regulation mechanisms of hair growth[J]. Current Problems in Dermatology, 11: 159-170. [22] Olivares C, Solano F.2009. New insights into the active site structure and catalytic mechanism of tyrosinase and its related proteins[J]. Pigment Cell & Melanoma Research, 22(6): 750-760. [23] Osawa M, Egawa G, Mak S S, et al.2005. Molecular characterization of melanocyte stem cells in their niche[J]. Development (Cambridge, England), 132(24): 5589-5599. [24] Park H J.2014. CARI ONE induces anagen phase of telogenic hair follicles through regulation of beta-catenin, stimulation of dermal papilla cell proliferation, and melanogenesis[J]. Journal of Dietary Supplements, 11(4): 320-333. [25] Sarangarajan R, Boissy R E.2001. Tyrp1 and oculocutaneous albinism type 3[J]. Pigment Cell Research, 14(6): 437-444. [26] Simon J D, Peles D N.2010. The red and the black[J]. Accounts of Chemical Research, 43(11): 1452-1460. [27] Slominski A, Tobin D J, Shibahara S, et al.2004. Melanin pigmentation in mammalian skin and its hormonal regulation[J]. Physiological Reviews, 84(4): 1155-1228. [28] Slominski A,Wortsman J, Plonka P M, et al.2005. Hair follicle pigmentation[J]. The Journal of investigative dermatology, 124(1): 13-21. [29] Sponenberg D P, Bellone R.1996. Equine color genetics, 4th edition[J]. Blackwell Publishers, 32(11): 1133-1140. [30] Stenn K S, Paus R.2001. Controls of hair follicle cycling[J]. Physiological Reviews, 81(1): 449-494. [31] Wu X, Zhang Y, Shen L, et al.2016. A 6-bp deletion in exon 8 and two mutations in introns of TYRP1 are associated with blond coat color in Liangshan pigs[J]. Gene, 578(1): 132-136. [32] Yamaguchi Y, Brenner M, Hearing V J.2007. The regulation of skin pigmentation[J]. The Journal of Biological Chemistry, 282(38): 27557-27561. [33] Yamaguchi Y, Passeron T, Hoashi T, et al.2008. Dickkopf 1 (DKK1) regulates skin pigmentation and thickness by affecting Wnt/beta-catenin signaling in keratinocytes[J]. FASEB Journal: Official Publication of the Federation of American Societies for Experimental Biology, 22(4): 1009-1020. [34] Zhang J, Liu F, Cao J, et al.2015. Skin transcriptome profiles associated with skin color in chickens[J]. PloS One, 10(6): e0127301. |
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