本研究旨在探讨RSPO_2和β-catenin对羊驼毛性状的作用。采用免疫组织化学、荧光定量PCR和Western blotting方法对RSPO_2和β-catenin在羊驼背部、腿部和耳部皮肤中基因和蛋白表达及蛋白定位进行研究。免疫组化结果显示:真皮乳头、毛囊基质、内根鞘、外根鞘、汗腺、表皮均可见RSPO_2的免疫阳性反应,并且主要出现在细胞质中,其中在内根鞘呈现强阳性;β-catenin在毛囊基质、内根鞘、外根鞘、汗腺、表皮内均出现免疫阳性反应。RT-PCR结果表明:RSPO_2在羊驼背部和腿部的表达量分别是耳部的7.633倍(P〈0.05)和5.602倍(P〈0.05),β-catenin在羊驼背部和腿部的表达量分别是耳部的3.689倍(P〈0.05)和2.067倍(P〉0.05)。Western blotting结果显示:羊驼皮肤提取物中存在与兔抗RSPO_2多克隆抗体和兔抗β-catenin多克隆抗体发生免疫阳性反应的蛋白条带,大小分别是26和86ku,羊驼皮肤平均蛋白表达量均为背部与耳部间差异显著(P〈0.05),其中,RSPO_2在腿部与耳部表达量差异显著(P〈0.05)。综上表明,RSPO_2和β-catenin与毛纤维的长度、弯曲度和细度呈正相关。
Alpaca fiber is the ideal material for the textile industry. Its length, fineness and curva- ture are different in the hack, leg and ear. In order to investigate the possible relationship between RSP02 and β-catenin with the characteristics of alpaca fleece, the expressions of RSP02 and β-catenin in back, leg and ear skins of alpaca were analyzed using immunohistochemistry, real-time PCR and Western blotting. The immunohistochemistry analysis demonstrated that the positive re- action of RSP02 was localized in the cytoplasmic of dermal papilla cells,hair follicle matrix,inner root sheath, outer root sheath, sweat glands and epidermis cells, in particular, the inner root sheath showed strongest positive reaction. The positive reaction of β-catenin was localized in hair follicle matrix, inner root sheath, outer root sheath, sweat glands and epidermis cells. Real-time PCR showed that the expression of RSP02 in back and leg skins were 7. 633 folds(P〈0.05) and 5. 602 folds(P〈0. 05), respectively, in comparison with that in ear skins. The expression of 19-catenin in back and leg skins were 3. 689 folds(P〈0.05) and 2. 067 folds(P〈0.05) ,respective-ly,in comparison with that in ear skins. Western blot analysis showed that the positive band of RSP02(26 ku) and ffcatenin (86 ku) were strongest in back skins and weakest in ear skins with significant difference (P〈0.05). The positive band of RSP02 in leg skins was significantly stron- ger than that in ear skins(P〈0.05). The results suggested that RSP02 and ffcatenin were posi- tively associated with regulating the length,curvature and fineness formation of alpaca fleece.