GPR143在绵羊皮肤组织中的表达及定位分析

G蛋白偶联受体143(G-protein coupled receptor143, GPR143)在黑素体的生物合成中起重要作用,本文旨在研究GPR143基因在不同毛色绵羊皮肤组织中的差异表达及定位,探索GPR143基因与毛色形成的相关性。通过qRT-PCR方法和免疫印迹方法分别检测不同毛色绵羊皮肤组织中GPR143基因mRNA水平和蛋白水平的表达差异;运用免疫荧光法对不同毛色绵羊皮肤组织中的GPR143基因进行定位并对结果进行光密度值分析。qRT-PCR结果显示,GPR143基因在黑色绵羊皮肤组织中mRNA相对表达量为白色绵羊的7.84倍,二者差异极显著(P<0.01);免疫印迹结果显示,黑色绵羊皮肤组织中GPR143蛋白表达量是白色绵羊的1.3倍,二者差异显著(P<0.05)。免疫荧光结果显示,GPR143蛋白的主要表达部位为绵羊皮肤组织毛囊外根鞘和表皮层,经光密度值分析后发现,GPR143在黑色绵羊皮肤毛囊外根鞘和表皮层的表达量显著高于白色绵羊。本研究结果表明不同毛色绵羊皮肤组织均能表达GPR143基因,但黑色绵羊皮肤组织中该基因的mRNA和蛋白水平都显著高于白色绵羊,说明GPR143的mRNA和蛋白在黑色绵羊皮肤组织中表达上调,在白色绵羊皮肤组织中表达下调。GPR143基因可能通过调控MITF水平和黑素体的数量、大小、运动和成熟进而参与绵羊毛色的形成过程。

Expression and localization of GPR143 in sheep skin

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.