藏绵羊不同毛色皮肤组织miRNA表达谱及靶基因分析  

Profiles of miRNAs and Target Gene Analysis with White and Black Skin Tissues of the Tibetan Sheep

作　　者：吴震洋[1,2] 唐晓惠 付玉华[2] 王昇 章程 李京津 余梅[2] 杜小勇[2,4] WU ZhenYang;TANG XiaoHui;FU YuHua;WANG Sheng;ZHANG Cheng;LI JingJin;YU Mei;DU XiaoYong(Tongren University, Tongren 554300, Guizhou;Huazhong Agricultural University/Key Lab of Animal Genetics, Breeding and Reproduction of Ministry Education, Wuhan 430070;Huazhong Agricultural University/Hubei Key Laboratory of Agricultural Bioinformatics, College of lnformatics, Wuhan 430070;XiZang Agriculture and Animal Husbandry College, Linzhi 860000, Tibet)

机构地区：[1]铜仁学院,贵州铜仁554300 [2]华中农业大学动物科技学院/农业动物遗传育种与繁殖教育部重点实验室,武汉430070 [3]西藏农牧学院,西藏林芝860000 [4]华中农业大学信息学院/农业生物信息湖北省重点实验室,武汉430070

出　　处：《中国农业科学》2018年第2期351-362,共12页Scientia Agricultura Sinica

基　　金：国家自然科学基金(31360534;31402040);中央高校基本科研业务费专项资金(2662015JC010);国家高技术研究发展计划(2013AA102506)

摘　　要：【目的】藏绵羊又称藏系绵羊,是中国三大粗毛羊品种之一,主要分布在西藏及其毗邻的高寒牧区,如青海、甘肃、四川、云南等地。藏绵羊由于其被毛纤维粗长,毛质细腻且保暖性好,是制造藏式地毯的上好原料。而毛色作为一种重要的经济性状,同时制约羊毛的经济价值。然而,目前对于绵羊毛色的分子调控机制尚不明确,以藏绵羊为研究对象,对其不同颜色皮肤组织(黑色和白色)进行转录组测序,旨在探讨miRNA在藏绵羊不同毛色皮肤组织转录后水平中发挥的作用及其可能参与的调控通路。【方法】选取2只具有黑白花毛色的健康藏绵羊,减去体侧被毛使皮肤完全裸露并用75%酒精消毒处理,屠宰后快速切取皮肤组织并保存在组织样品保存液中防止RNA降解,提取RNA后通过miRNA测序和生物信息学分析,获得藏绵羊不同毛色(黑色和白色)皮肤组织miRNA表达谱,筛选组织差异表达miRNA并预测相关靶基因。通过靶基因分析,获得与调控藏绵羊毛色性状可能相关的信号通路。【结果】黑色和白色皮肤组织共获得85.76兆条原始读长,经过滤后得到85.08兆条clean reads;共鉴定出334个已知的miRNA和59个新的miRNA;从中获得23个差异表达miRNA,其中14个差异表达miRNA在白色皮肤组织中表达显著上调,9个表达下调。miR-2284b和miR-744仅在藏绵羊白色皮肤中表达,而miR-23b、miR-411a-5p、miR-30c、miR-423-3p和miR-324-5p则仅在藏绵羊黑色皮肤中表达,但表达量相对较低。表达量最高的miRNA为miR-10a,而倍数差异最大的为miR-23b。其中,miR-10a可参与调节多种信号通路,其作用涉及增殖、分化、凋亡、细胞粘附等各个方面,目前并无调控绵羊毛色方面的相关报道,但其功能及作用机制的研究仍在不断扩展。miR-23b则与Wnt、Notch等信号通路有关,这些信号通路中的部分基因也和黑色素的生成有密切关系。结合倍数差异和miRNA表达【Objective】 Tibetan sheep is one of three coarse wool sheep breeds in China, distributing mainly in Tibet and its adjacent alpine pastoral areas, such as Qinghai, Gansu, Sichuan, Yunnan. The wool of Tibetan sheep, felt smooth and warm duo to the long and thick fibers, is good raw material for manufacture of Tibetan carpet. Coat color is an important economic traits. However, at present, the mechanism of molecular regulation of sheep coat color is not clear. In this study, the transcriptome of skin tissues of different color（black and white） was sequenced with a aim to explore the role of miRNAs at post-transcriptional levels in different coat color skin tissues and possible regulatory pathways. 【Method】Healthy sheep with black and white coat color was sacrificed to provide skin tissues for RNA extraction and miRNA analysis. Through miRNA sequencing and bioinformatics analysis, miRNA expression profiles of skin tissues of black and white wool color were obtained. Then the differentially expressed miRNAs were screened and the related target genes were predicted. Through the target gene analysis, the signal pathway which is related to Tibetan coat color skin traits were proposed. 【Result】 A total of 85.76 million original reads and 85.08 million clean reads were obtained from the analyzed tissues. Among the clean reads were 334 known miRNAs and 59 newly identified miRNAs and 23 of them were differentially expressed between white and black color tissues. In the 23 differentially expressed miRNAs, 14 and 9 were up-regulated and down-regulated in white skin tissues respectively. miR-2284 b and miR-744 were the type that is expressed only in white Tibetan sheep skin, and miR-23 b, miR-411 a-5 p, miR-30 c, miR-423-3 p, miR-324-5 p were the miRNAs that expressed only in black skin. Among them, miR-10 a can participate in the regulation of a variety of signaling pathways, involving proliferation, differentiation, apoptosis, cell adhesion. Mi R-23 b is associated with Wnt, Notch and other signal pathways, wh

关 键 词：藏绵羊 皮肤组织 高通量测序 MIRNA 

分 类 号：S826[农业科学—畜牧学]