miR-411a-3p抑制羊驼黑色素细胞黑色素的生成
发布时间：2018-11-12 13:12
【摘要】：microRNA-411a-3p(miR-411a-3p)在不同毛色羊驼皮肤中差异表达,提示其可能在毛色形成中起调控作用。为证明miR-411a-3p在羊驼皮毛黑色素形成中的作用,本研究通过生物信息学预测及靶基因搜索,发现胰岛素样生长因子1受体(IGF1R)是miR-411a-3p的靶基因,继而构建了含Igf1r mRNA 3'-UTR的荧光素酶报告基因载体。报告基因转染结合报告酶活性测定证明,Igf1r是miR-411a-3p的靶基因。原位杂交显示,miR-411a-3p主要在羊驼黑色素细胞胞质中表达,提示其可能在黑色素细胞中具有重要作用。qRT-PCR揭示,棕色和黑色羊驼皮肤中miR-411a-3p表达量明显低于白色羊驼。qRT-PCR联合蛋白质印迹分析显示,在羊驼黑色素细胞过表达miR-411a-3p,伴随Igf1r下调,小眼畸形相关转录因子(Mitf)依赖的毛色基因酪氨酸酶(Tyr)、酪氨酸酶相关蛋白-2(Tyrp2)及黑色素表达水平明显下调。上述结果提示,miR-411a-3p可通过靶向抑制Igf1r负调控羊驼黑色素黑色素合成。该结果将加深我们对羊驼毛色形成机制的认识。
[Abstract]:The differential expression of microRNA-411a-3p (miR-411a-3p) in the skin of alpaca with different color suggests that miR-411a-3p may play a regulatory role in the formation of wool color. In order to demonstrate the role of miR-411a-3p in the formation of melanin in alpaca fur, we found that insulin like growth factor 1 receptor (IGF1R) is the target gene of miR-411a-3p by bioinformatics prediction and target gene search. Then the luciferase reporter gene vector containing Igf1r mRNA 3'-UTR was constructed. The reporter gene transfection combined with reporter enzyme activity showed that Igf1r was the target gene of miR-411a-3p. In situ hybridization showed that miR-411a-3p was mainly expressed in the cytoplasm of melanocytes of alpaca, suggesting that miR-411a-3p may play an important role in melanocytes. The expression of miR-411a-3p in brown and black alpaca skin was significantly lower than that in white alpaca. QRT-PCR combined with Western blot analysis showed that the overexpression of miR-411a-3p, was accompanied by down-regulation of Igf1r in the melanocytes of alpaca. The expression of tyrosinase (Tyr), tyrosinase protein-2 (Tyrp2) and melanin were down-regulated by (Mitf) dependent transcription factor. These results suggest that miR-411a-3p can inhibit the negative regulation of melanin synthesis in alpaca by targeting Igf1r. This result will deepen our understanding of the formation mechanism of alpaca coat color.
【作者单位】： 山西农业大学动物科技学院;
【基金】：国家大学生创新创业训练项目(No.201710113025) 山西省“三晋学者”人才专项资助~~
【分类号】：S829.9
本文编号：2327211
[Abstract]:The differential expression of microRNA-411a-3p (miR-411a-3p) in the skin of alpaca with different color suggests that miR-411a-3p may play a regulatory role in the formation of wool color. In order to demonstrate the role of miR-411a-3p in the formation of melanin in alpaca fur, we found that insulin like growth factor 1 receptor (IGF1R) is the target gene of miR-411a-3p by bioinformatics prediction and target gene search. Then the luciferase reporter gene vector containing Igf1r mRNA 3'-UTR was constructed. The reporter gene transfection combined with reporter enzyme activity showed that Igf1r was the target gene of miR-411a-3p. In situ hybridization showed that miR-411a-3p was mainly expressed in the cytoplasm of melanocytes of alpaca, suggesting that miR-411a-3p may play an important role in melanocytes. The expression of miR-411a-3p in brown and black alpaca skin was significantly lower than that in white alpaca. QRT-PCR combined with Western blot analysis showed that the overexpression of miR-411a-3p, was accompanied by down-regulation of Igf1r in the melanocytes of alpaca. The expression of tyrosinase (Tyr), tyrosinase protein-2 (Tyrp2) and melanin were down-regulated by (Mitf) dependent transcription factor. These results suggest that miR-411a-3p can inhibit the negative regulation of melanin synthesis in alpaca by targeting Igf1r. This result will deepen our understanding of the formation mechanism of alpaca coat color.
【作者单位】： 山西农业大学动物科技学院;
【基金】：国家大学生创新创业训练项目(No.201710113025) 山西省“三晋学者”人才专项资助~~
【分类号】：S829.9
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