SOX5影响小鼠皮肤黑色素细胞MITF-M表达和黑色素生成

摘要

To investigate the function of SOX5 in mouse skins and the regulation of hair color formation,the differential expression of SOX5 in skins of different coat color mouse skin was examined.In this study,C57BL/6 strains of black,brown and gray mice were selected each of the 3 for 12 days after birth,randomly.Quantitative real-time PCR (qRT-PCR),Western blotting and immunohistochemistry methods were used in this study to analyze the expression of SOX5;we also constructed the eukaryotic expression vector of SOX5 and transfected the mouse melanocytes to detect the expression level of the pigmentation-related genes,and measured the melanin content.The results showed that:1) SOX5 could be expressed in mouse skin with different hair color.The mRNA and protein expression levels of SOX5 in black and gray mouse skin were higher than that in brown mouse skin with very significantly difference (P＜0.01).And the main expression site of SOX5 was hair follicle outer root sheath.2) Compared with empty vector group(Vector GFP),the mRNA and protein expression levels of MITF-M,TYR,TYRP1,TYRP2,PMEL andOA1 were increased with significant difference (P＜0.05) and very significantly difference (P＜ 0.01) in SOX5 transfected group;the content of melanin was significantly increased with very significantly difference (P＜0.01) in SOX5 transfected group.3) MITF-M had a negative feedback on SOX5 by overexpression of MITF-M.These suggest that SOX5 affect the formation of pigments and then involve in the formation of coat color by regulating MITF-M.%旨在探讨SOX5在小鼠皮肤毛色中的作用.本研究随机选取出生后12d的C57BL/6品系黑色、棕色、灰色小鼠各3只,采用实时荧光定量PCR、Western blotting和免疫组织化学方法对SOX5在黑色、棕色和灰色小鼠皮肤中的表达差异进行分析;并构建SOX5真核表达载体;运用体外转染的方法,转染小鼠黑色素细胞并检测与色素形成相关基因的表达水平,同时测定黑色素含量.结果表明:1)SOX5在不同毛色小鼠皮肤中均有表达,SOX5在黑色和灰色小鼠皮肤中mRNA和蛋白相对表达量高于棕色小鼠皮肤,均差异极显著(P＜0.01);且其主要表达部位为毛囊外根鞘.2)转染组与空载组相比,MITF-M、TYR、TYRP1、TYRP2、PMEL、OA1表达量均显著(P＜0.05)或极显著(P＜0.01)升高,黑色素含量也极显著增加(P＜0.01).3)过表达MITF-M对SOX5存在负反馈作用.综上表明,SOX5通过调控MITF-M影响色素的生成进而参与毛色的形成.