内皮素3在不同毛色绵羊皮肤的表达与定位

EDN3 Expression and Localization in Sheep Skin with Different Coat Color

旨在探索内皮素3(Endothelin 3,EDN3)在不同毛色绵羊皮肤的表达差异以及对毛色的影响。以不同毛色绵羊为研究对象,通过荧光定量PCR技术分析EDN3基因在不同毛色绵羊皮肤的表达量,通过免疫组织化学和ELISA方法对EDN3蛋白在不同毛色绵羊皮肤中的定位和表达进行研究。1)qRT-PCR结果显示,EDN3在白色绵羊皮肤的相对表达量为5.540 6±0.030 7,在黑色绵羊皮肤的相对表达量为1.005 8±0.062 0。黑白花色绵羊的白色区和黑色区的相对表达量分别为4.341 9±0.087 7和1.150 1±0.005 3。2)ELISA结果显示,EDN3在白色绵羊皮肤中的表达量为128.424 7±2.223 4,在黑色绵羊皮肤的相对表达量为25.114 4±3.248 3。在黑白花色绵羊白色皮肤中的表达量为93.945 3±7.562 2,黑色皮肤的表达量为28.606 0±9.295 9。3)免疫组织化学显示,EDN3在绵羊皮肤毛囊的毛基质、内外毛根鞘、毛乳头等区域均有表达。综上表明,EDN3在不同毛色绵羊皮肤中均可正常表达,且表达量存在显著差异。EDN3是维持黑色素细胞存在不可缺少的基因,但不影响绵羊黑白花的形成。

To explore the expression of endothelin 3in different coat color of sheep skin and the effect of EDN3 on the sheep coat color,the experiment analyzed the mRNA and protein expression level of EDN3 in sheep skin of different coat colors in different coat color ship by real-time quantitative PCR,ELISA and immunohistochemistry.1)qRT-PCR showed that the relative expressive of EDN3 mRNA in white sheep skin was 5.540 6±0.030 7,while in black was 1.005 8±0.062 0.The gene expression quantity of EDN3 in white and black skin tissue of the same individuality was 4.341 9±0.087 7and 1.150 1±0.005 3.2)ELISA results showed that the expression of EDN3 protein in white and black were 128.424 7±2.223 4and 25.114 4±3.248 3.The protein expression of EDN3 in white and black skin of the same individuality were 93.945 3±7.562 2and 28.606 0±9.295 9.3)The EDN3 was located in the matrix,outer root sheath and dermal papilla of hair follicular by immunohischemistry.Experiments showed that the EDN3 expressed normally in black and white sheep skin,and the expression quantity had significant differences.The research indicated that EDN3 was indispensable to maintain the presence of the melanocyte,and didn't affect the formation of friesian sheep.