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作 者:张丹瑾[1,2] 谢建山[1] 申艳[1] 范瑞文[1] 聂瑞强[1] 杨玉静[1] 于秀菊[1] 段力升 曹靖[3] 董常生[1]
机构地区:[1]山西农业大学动物科技学院,山西太谷030801 [2]山西医科大学晋祠学院,山西太原030025 [3]北京农业职业学院,北京102442
出 处:《中国生物化学与分子生物学报》2015年第10期1109-1116,共8页Chinese Journal of Biochemistry and Molecular Biology
基 金:国家高科技研究发展计划(863计划;No.2013AA102506);公益性行业(农业)科研专项(No.201303119);农业部"948"项目(No.2011-Z33)资助~~
摘 要:突触融合蛋白17(STX17)是一种囊泡蛋白,参与细胞中物质的运输.为研究Stx17在不同毛色皮肤中是否存在差异表达及明确它在毛囊中的定位,进行了普通PCR、real-time PCR、免疫组化和蛋白免疫印迹实验对小鼠皮肤组织和体外培养黑素细胞的Stx17基因及蛋白的检测.普通PCR检测得出小鼠皮肤和黑色素细胞总RNA有Stx17 CDS区序列的表达;荧光定量检测显示,在白、灰、黑3种组织中Stx17均有表达,在灰色腹部表达量最高,是黑色皮肤的1.682倍,昆明鼠白色皮肤中表达量最低,是黑色皮肤的0.115倍;皮肤组织免疫组化结果显示,STX17表达于毛囊的上皮根鞘,且毛囊上段和中段表达量高于下段,黑色素细胞的免疫组化分析得出,STX17在黑色素细胞的细胞质和细胞膜上均有表达;蛋白免疫印迹结果显示,在白色、灰色和黑色皮肤均有STX17蛋白阳性条带且灰色皮肤中表达量最高,黑色皮肤次之,白色皮肤中表达量是最低的,这与荧光定量检测结果一致,体外培养的小鼠黑色素细胞中也有STX17蛋白阳性条带.实验结果表明,小鼠Stx17基因在皮肤组织、毛囊角化细胞以及黑色素细胞中均有表达,Stx17可能参与毛色的形成,且在小鼠腹部毛色变浅中起到了一定的作用.Syntaxin17( STX17) is a vesicle protein involved in the transporting of cellular substances.To investigate its localization in hair follicles and determine whether it differentially expressed in the skin of different coat colors,mouse skin tissues and cultured mouse melanocytes were analyzed by conventional or real-time PCR,immunohistochemistry and Western blotting. The results showed that Stx17 CDS were detected in the total RNAs from white,grey and black skins. The highest expression level in grey abdomen was 1. 682 times more than in black skin; the lowest in white Kunming mice skin was 0. 115.Immunohistochemistry results showed STX17 was expressed in the whole hair follicles in black mice,especially at the upper and middle parts. In gray skin hair follicle cross sections,STX17 was expressed in the epithelial root sheath within melanocytes. STX17 proteins was detected in white,black,and gray skins with increasing levels in immunoblots,which was consistent with the real-time PCR results. In melanocyte in vitro cultures,STX17 was detected positively. We concluded that Stx17 was expressed in mouse skin and localization in keratinocyte and melanocyte in hair follicle.
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