Yin Zhihong, Ma Zhisheng, Wang Siting, Hao Shitong, Liu Xinyou, Pang Quanhai, Wang Xinzhuang
Postdoctoral Research Base, College of Veterinary Medicine, Henan Agricultural University, Zhengzhou 450046, China.
College of Animal Science and Veterinary Medicine, Henan Institute of Science and Technology, Xinxiang 453003, China.
Anim Biosci. 2023 Sep;36(9):1367-1375. doi: 10.5713/ab.23.0111. Epub 2023 Jun 23.
Pigment production and distribution are controlled through multiple proteins, resulting in different coat color phenotypes of sheep.
The expression distribution of vimentin (VIM) and transthyretin (TTR) in white and black sheep skins was detected by liquid chromatography-electrospray ionization tandem MS (LC-ESI-MS/MS), gene ontology (GO) statistics, immunohistochemistry, Western blot, and quantitative real time polymerase chain reaction (qRT-PCR) to evaluate their role in the coat color formation of sheep.
LC-ESI-MS/MS results showed VIM and TTR proteins in white and black skin tissues of sheep. Meanwhile, GO functional annotation analysis suggested that VIM and TTR proteins were mainly concentrated in cellular components and biological process, respectively. Further research confirmed that VIM and TTR proteins were expressed at significantly higher levels in black sheep skins than in white sheep skins by Western blot, respectively. Immunohistochemistry notably detected VIM and TTR in hair follicle, dermal papilla, and outer root sheath of white and black sheep skins. qRT-PCR results also revealed that the expression of VIM and TTR mRNAs was higher in black sheep skins than in white sheep skins.
The expression of VIM and TTR were higher in black sheep skins than in white sheep skins and the transcription and translation were unanimous in this study. VIM and TTR proteins were expressed in hair follicles of white and black sheep skins. These results suggested that VIM and TTR were involved in the coat color formation of sheep.
色素的产生和分布由多种蛋白质控制,从而导致绵羊出现不同的毛色表型。
采用液相色谱-电喷雾电离串联质谱(LC-ESI-MS/MS)、基因本体论(GO)统计、免疫组织化学、蛋白质免疫印迹法以及实时定量聚合酶链反应(qRT-PCR)检测波形蛋白(VIM)和转甲状腺素蛋白(TTR)在白色和黑色绵羊皮肤中的表达分布,以评估它们在绵羊毛色形成中的作用。
LC-ESI-MS/MS结果显示绵羊白色和黑色皮肤组织中存在VIM和TTR蛋白。同时,GO功能注释分析表明,VIM和TTR蛋白分别主要集中在细胞成分和生物学过程中。进一步研究通过蛋白质免疫印迹法证实,VIM和TTR蛋白在黑色绵羊皮肤中的表达水平分别显著高于白色绵羊皮肤。免疫组织化学显著检测到白色和黑色绵羊皮肤的毛囊、真皮乳头和外根鞘中有VIM和TTR。qRT-PCR结果还显示,VIM和TTR mRNA在黑色绵羊皮肤中的表达高于白色绵羊皮肤。
本研究中,VIM和TTR在黑色绵羊皮肤中的表达高于白色绵羊皮肤,且转录和翻译一致。VIM和TTR蛋白在白色和黑色绵羊皮肤的毛囊中表达。这些结果表明,VIM和TTR参与了绵羊毛色的形成。