Tian Xue, Meng Xiaolin, Wang Liangyan, Song Yunfei, Zhang Danli, Ji Yuankai, Li Xuejun, Dong Changsheng
College of Fisheries, Henan Normal University, Xinxiang 453007, PR China.
BGI-Shenzhen, Shenzhen 518083, PR China.
Gene. 2015 Jan 25;555(2):88-94. doi: 10.1016/j.gene.2014.10.029. Epub 2014 Oct 20.
Slc7a11 encoding solute carrier family 7 member 11 (amionic amino acid transporter light chain, xCT), has been identified to be a critical genetic regulator of pheomelanin synthesis in hair and melanocytes. To better understand the molecular characterization of Slc7a11 and the expression patterns in skin of white versus brown alpaca (lama paco), we cloned the full length coding sequence (CDS) of alpaca Slc7a11 gene and analyzed the expression patterns using Real Time PCR, Western blotting and immunohistochemistry. The full length CDS of 1512bp encodes a 503 amino acid polypeptide. Sequence analysis showed that alpaca xCT contains 12 transmembrane regions consistent with the highly conserved amino acid permease (AA_permease_2) domain similar to other vertebrates. Sequence alignment and phylogenetic analysis revealed that alpaca xCT had the highest identity and shared the same branch with Camelus ferus. Real Time PCR and Western blotting suggested that xCT was expressed at significantly high levels in brown alpaca skin, and transcripts and protein possessed the same expression pattern in white and brown alpaca skins. Additionally, immunohistochemical analysis further demonstrated that xCT staining was robustly increased in the matrix and root sheath of brown alpaca skin compared with that of white. These results suggest that Slc7a11 functions in alpaca coat color regulation and offer essential information for further exploration on the role of Slc7a11 in melanogenesis.
编码溶质载体家族7成员11(阴离子氨基酸转运体轻链,xCT)的Slc7a11,已被确定为毛发和黑素细胞中褐黑素合成的关键基因调节因子。为了更好地了解Slc7a11的分子特征以及白色和棕色羊驼(小羊驼)皮肤中的表达模式,我们克隆了羊驼Slc7a11基因的全长编码序列(CDS),并使用实时PCR、蛋白质印迹和免疫组织化学分析了其表达模式。1512bp的全长CDS编码一个503个氨基酸的多肽。序列分析表明,羊驼xCT含有12个跨膜区域,与其他脊椎动物中高度保守的氨基酸通透酶(AA_permease_2)结构域一致。序列比对和系统发育分析表明,羊驼xCT具有最高的同源性,与野生骆驼处于同一分支。实时PCR和蛋白质印迹表明,xCT在棕色羊驼皮肤中的表达水平显著较高,并且在白色和棕色羊驼皮肤中转录本和蛋白质具有相同的表达模式。此外,免疫组织化学分析进一步表明,与白色羊驼相比,棕色羊驼皮肤的基质和根鞘中xCT染色明显增加。这些结果表明,Slc7a11在羊驼毛色调节中发挥作用,并为进一步探索Slc7a11在黑色素生成中的作用提供了重要信息。