蒙古斑点马黑、白毛色区域皮肤中相关基因的表达研究

doi:10.11843/j.issn.0366-6964.2022.01.010

畜牧兽医学报 ›› 2022, Vol. 53 ›› Issue (1): 100-111.doi: 10.11843/j.issn.0366-6964.2022.01.010

蒙古斑点马黑、白毛色区域皮肤中相关基因的表达研究

白东义¹, 拉希玛¹, 赵若阳¹, 陶娜拉³, 韩海格¹, 丁文淇¹, 贾紫洁¹, 刘慧莹¹, 王文兴², 黄博光², 芒来^1*

1. 内蒙古农业大学动物科学学院内蒙古自治区马属动物遗传育种与繁殖重点实验室农业农村部马属动物遗传育种与繁殖科学观测实验站内蒙古农业大学马属动物研究中心, 呼和浩特 010018;
2. 内蒙古中蕴马产业集团, 锡林浩特 026000;
3. 锡林郭勒盟镶黄旗农牧和科技局, 镶黄旗 013250

收稿日期:2021-04-06 出版日期:2022-01-23 发布日期:2022-01-26
通讯作者: 芒来,主要从事马属动物遗传育种与繁殖研究,E-mail:dmanglai@163.com
作者简介:白东义(1983-),男,蒙古族,内蒙古宁城人,副教授,博士,主要从事马属动物遗传育种与繁殖研究,E-mail:baidongyi1983@163.com;拉希玛(1994-),女,蒙古族,内蒙古锡林郭勒盟正蓝旗人,硕士生,主要从事马属动物遗传育种与繁殖研究,E-mail:361644344@qq.com。
基金资助:
国家自然科学基金（31960657）；自治区科技重大专项（ZD20190039）；内蒙古农业大学动物科学学院标志性成果专项资金项目（BZCG201901）；内蒙古农业大学动物科学学院青年基金项目（QN202006）

Study on the Expression of Related Genes in the Skin of Mongolian Spotted Horses in Black and White Coat Regions

BAI Dongyi¹, LA Xima¹, ZHAO Ruoyang¹, TAO Nala³, HAN Haige¹, DING Wenqi¹, JIA Zijie¹, LIU Huiying¹, WANG Wenxing², HUANG Boguang², MANG Lai^1*

1. Equine Research Center, Scientific Observing and Experimental Station of Equine Genetics/Breeding and Reproduction, Ministry of Agriculture and Rural Affairs, Inner Mongolia Key Laboratory of Equine Genetics/Breeding and Reproduction, College of Animal Science, Inner Mongolia Agricultural University, Hohhot 010018, China;
2. Inner Mongolia Zhong Yun Horse Industry Group, Xilinhot 026000, China;
3. Xilingol League Xianghuangqi Agriculture, Animal Husbandry and Science and Technology Bureau, Xianghuangqi 013250, China

Received:2021-04-06 Online:2022-01-23 Published:2022-01-26

摘要/Abstract

摘要： 本试验以蒙古斑点马为研究对象，分别对蒙古斑点马体躯白色区域和黑色区域皮肤的表型和差异表达基因进行分析并验证，试图解析蒙古斑点马毛色形成的分子机制，为今后保护和开发利用蒙古斑点马奠定基础。选择蒙古斑点马体躯白色和黑色区域皮肤制作组织切片，HE染色后在显微镜下观察其表型差异；对白色和黑色区域皮肤组织进行转录组测序，比较差异表达mRNAs；将筛选到的典型基因通过免疫荧光试验和实时荧光定量试验，对转录组测序结果进一步验证。结果显示，蒙古斑点马黑色皮肤组织和白色皮肤组织中的色素分布有明显的差异，黑色素细胞主要分布于黑色皮肤组织表皮和毛囊毛球部位，而白色皮肤组织相应的位置未发现有黑色素沉积。740个基因的表达量在黑白两个不同颜色皮肤组织中存在显著差异（P<0.05），其中，109个基因在蒙古斑点马黑色皮肤组织表达量较高，215个基因在蒙古斑点马白色皮肤组织表达量较高。差异表达mRNAs主要富集到了黑色素生成等通路，其中TYR、TYRP1、DCT、PAX3、DAPL1等基因与毛色形成相关。在蒙古斑点马毛囊中，TYR和TYRP1蛋白在毛囊的毛球部位不表达。DCT在毛囊毛球的毛母质部位表达。因此，蒙古斑点马的毛色形成主要由黑色素形成相关的基因的表达量所决定。

关键词: 蒙古斑点马, 皮肤组织, 差异表达mRNA

Abstract: In this study, phenotypes and differentially expressed genes in the white and black regions of the body of Mongolian spotted horse were analyzed and verified, in an attempt to elucidate the molecular mechanism of hair color formation of Mongolian spotted horse, and lay a foundation for future protection, development and utilization of Mongolian spotted horse. First, skin tissue sections of the white and black regions of the body of Mongolian spotted horse were made and stained by HE staining method and phenotypic differences were observed under a microscope. Secondly, transcriptome sequencing was performed on the skin of white and black regions to compare the differentially expressed mRNAs. Finally, the selected typical genes were tested by immunofluorescence immunohistochemistry and qRT-PCR, the results of transcriptome sequencing were further verified. The results showed that there were significant differences in the pigment distribution between the black and white skin tissues of Mongolian spotted horses. Melanocytes were mainly distributed in the epidermis and hair follicle parts of the black skin tissues, while no melanin deposition was found in the corresponding positions of the white skin tissues. The expression levels of 740 genes were significantly different between black and white skin tissues (P<0.05), among which 109 genes were highly expressed in black skin tissue of Mongolian spotted horse, and 215 genes were highly expressed in white skin tissue of Mongolian spotted horse. Differentially expressed mRNAs were mainly enriched in melanin production pathway contained TYR, TYRP1, DCT, PAX3 and DAPL1, etc.genes. TYR and TYRP1 proteins were not expressed in the hair bulb of the hair follicle of Mongolian spotted horse. DCT was expressed in the mother material of hair follicle. The hair color formation of Mongolian spotted horse is mainly determined by the expression of genes related to melanin formation.

Key words: Mongolian spotted horse, skin tissue, differentially expressed mRNA

中图分类号:

S821.81.2

白东义, 拉希玛, 赵若阳, 陶娜拉, 韩海格, 丁文淇, 贾紫洁, 刘慧莹, 王文兴, 黄博光, 芒来. 蒙古斑点马黑、白毛色区域皮肤中相关基因的表达研究[J]. 畜牧兽医学报, 2022, 53(1): 100-111.

BAI Dongyi, LA Xima, ZHAO Ruoyang, TAO Nala, HAN Haige, DING Wenqi, JIA Zijie, LIU Huiying, WANG Wenxing, HUANG Boguang, MANG Lai. Study on the Expression of Related Genes in the Skin of Mongolian Spotted Horses in Black and White Coat Regions[J]. Acta Veterinaria et Zootechnica Sinica, 2022, 53(1): 100-111.

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蒙古斑点马黑、白毛色区域皮肤中相关基因的表达研究

Study on the Expression of Related Genes in the Skin of Mongolian Spotted Horses in Black and White Coat Regions

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