Yin Zhihong, Zhao Xin, Wang Zhun, Wang Lirui, Li Zhen, Bai Rui, Zhao Min, Pang Quanhai
College of Animal Science and Veterinary Medicine, Shanxi Agricultural University, Taigu, Shanxi 030801, PR China.
Department of Medicine, University of California, San Diego, La Jolla, CA 92093, USA.
Acta Histochem. 2016 Mar;118(2):170-5. doi: 10.1016/j.acthis.2015.12.008. Epub 2016 Jan 6.
Guanine nucleotide-binding protein subunit alpha-s (Gnαs) and guanine nucleotide-binding protein subunit alpha-11 (Gnα11) play an important role in coat color formation. To improve our understanding of Gnαs and Gnα11 expression levels and pattern in the skin of black sheep (Ovis aries) compared with white sheep, we analyzed the expression levels through quantitative real time PCR (qPCR) and Western blot, immunohistochemistry and immunofluorescence. qPCR and Western blot results suggested that Gnαs and Gnα11 were significantly expressed at high levels in black sheep skin compared with the white sheep skin. Gnα11 expression was higher than Gnαs expression in both skin colors, transcripts and protein exhibited the same expression pattern in white and black sheep skins. Immunohistochemical results revealed that Gnαs and Gnα11 were localized in the outer root sheath of hair follicle in sheep. Furthermore, the expression of Gnα11 in outer root sheath of hair follicle was stronger than that in Gnαs. Immunofluorescencence further demonstrated that signals of Gnαs and Gnα11 were detected in outer root sheath and hair papilla. These results provide a novel insight into the role of Gnαs and Gnα11 in the regulation of sheep coat color.
鸟嘌呤核苷酸结合蛋白α-s亚基（Gnαs）和鸟嘌呤核苷酸结合蛋白α-11亚基（Gnα11）在毛色形成中起重要作用。为了更好地理解与白羊相比，黑羊（绵羊）皮肤中Gnαs和Gnα11的表达水平及模式，我们通过实时定量聚合酶链反应（qPCR）、蛋白质免疫印迹、免疫组织化学和免疫荧光分析了其表达水平。qPCR和蛋白质免疫印迹结果表明，与白羊皮肤相比，Gnαs和Gnα11在黑羊皮肤中显著高表达。两种毛色皮肤中Gnα11的表达均高于Gnαs，转录本和蛋白质在白羊和黑羊皮肤中呈现相同的表达模式。免疫组织化学结果显示，Gnαs和Gnα11定位于绵羊毛囊外根鞘。此外，Gnα11在毛囊外根鞘中的表达强于Gnαs。免疫荧光进一步证明，在毛囊外根鞘和毛乳头中检测到Gnαs和Gnα11的信号。这些结果为Gnαs和Gnα11在绵羊毛色调控中的作用提供了新的见解。
