Chen Tian-zhi, Zhao Bing-ling, Liu Yu, Zhao Yuan-yuan, Wang Hai-dong, Fan Rui-wen, Wang Peng-chao, Dong Chang-sheng
College of Animal Science and Veterinary Medicine, Shanxi Agricultural University, Taigu 030801, China.
College of Animal Science and Technology, Northeast Agricultural University, Harbin 150030, China.
Yi Chuan. 2016 Jul 20;38(7):658-665. doi: 10.16288/j.yczz.16-068.
G-protein coupled receptor143 (GPR143) plays an important role in melanogenesis. In this study, we investigated the expression pattern and localization of GPR143 in skin of sheep with different coat colors and explored the correlation between GPR143 gene and coat color. The mRNA level and protein level of GPR143 in skin of sheep with different coat colors were detected by qRT-PCR and immunoblotting separately while the localization of GPR143 in sheep skin was detected by immunofluorescence assay following optical density analysis. The qRT-PCR results showed that the relative expression level of GPR143 mRNA in black sheep skin was 7.84 times of that in white sheep skin (P<0.01). Immunoblotting results demonstrated that the expression level of GPR143 protein in black sheep skin was 1.30 times of that in white sheep skin (P<0.05). Immunofluorescence assay revealed that GPR143 was primarily expressed in the outer root sheath of hair follicles and epidermal skin tissue. Optical density analysis showed that expression levels of GPR143 in the outer root sheath and epidermis of black sheep skin were significantly higher than that of white sheep skin. Our studies demonstrated that GPR143 is expressed in skin of sheep with different coat colors. However, the mRNA and protein levels of GPR143 in black sheep skin are significantly higher than that in white sheep skin, indicating that GPR143 mRNA and protein levels are upregulated in skin of black sheep while downregulated in skin of white sheep. GPR143 may participate in the formation of coat color by regulating the level of MITF and the number, size, motility and maturation of the melanosome.
G蛋白偶联受体143（GPR143）在黑色素生成中起重要作用。在本研究中，我们调查了不同毛色绵羊皮肤中GPR143的表达模式和定位，并探讨了GPR143基因与毛色之间的相关性。分别采用qRT-PCR和免疫印迹法检测不同毛色绵羊皮肤中GPR143的mRNA水平和蛋白水平，同时采用免疫荧光分析法检测GPR143在绵羊皮肤中的定位，并进行光密度分析。qRT-PCR结果显示，黑羊皮肤中GPR143 mRNA的相对表达水平是白羊皮肤中的7.84倍（P<0.01）。免疫印迹结果表明，黑羊皮肤中GPR143蛋白的表达水平是白羊皮肤中的1.30倍（P<0.05）。免疫荧光分析显示，GPR143主要表达于毛囊外根鞘和表皮皮肤组织。光密度分析表明，黑羊皮肤外根鞘和表皮中GPR143的表达水平显著高于白羊皮肤。我们的研究表明，GPR143在不同毛色绵羊的皮肤中均有表达。然而，黑羊皮肤中GPR143的mRNA和蛋白水平显著高于白羊皮肤，这表明GPR143的mRNA和蛋白水平在黑羊皮肤中上调，而在白羊皮肤中下调。GPR143可能通过调节小眼畸形相关转录因子（MITF）的水平以及黑素小体的数量、大小、运动性和成熟度来参与毛色的形成。
