羊驼黑色素皮质素受体1在绵羊黑色素细胞中的表达促进黑色素的生成（英文）

Expression of Alp-MC1R in Sheep Melanocyte Stimulates Melanin Production

已知黑色素皮质素受体1(melanocortin-1 receptor,MC1R)是调控哺乳动物毛色的主要基因之一。羊驼是一种能生产纤维的家畜,有22种天然毛色。为了研究是否可以将羊驼的毛色基因应用到绵羊中,本文克隆了羊驼MC1R,发现羊驼MC1R与绵羊的相似性为88. 05%。将表达羊驼MC1R的质粒转染到绵羊黑色素细胞中,MC1R、小眼畸形相关转录因子(microphthalmia-associated transcription factor,MITF)、酪氨酸酶(tyrosinase,TYR)、酪氨酸酶相关蛋白2 (tyrosinase-related protein 2,TYRP2)在转录水平的表达量与NC组相比具有显著上升趋势,分别是对照组的3. 193±0. 063倍、2. 021±0. 129倍、3. 218±0. 056倍、3. 140±0. 108倍(P<0. 01)。Western印迹检测MITF、TYR、TYRP2在蛋白质水平的表达分别是对照组的1. 852±0. 117倍、3. 537±0. 13倍、2. 045±0. 163倍,具有显著差异(P<0. 01)。此外,绵羊黑色素细胞中羊驼MC1R的表达造成黑色素合成显著增加,是NC组的1. 715±0. 014倍(P<0. 01)。本研究结果表明,羊驼MC1R可用于增加绵羊黑色素生成过程中某些毛色基因的表达,为羊驼提供了可能的应用策略。

Melanocortin-1 receptor(MC1 R)is known to be one of the main genes to control coat color in mammals.Alpaca,fiber-producing domestic animal,has 22 kinds of natural hair colors.To investigate whether hair color gene of alpaca can be applied to sheep,MC1 R from Alpaca(Alp-MC1 R)was cloned and results showed that the similarity between Alp-MC1 R and sheep was 88.05%similarity.The plasmid expressing Alp-MC1 R was transfected into sheep melanocytes and a significant increase was found in mRNA expression of MC1 R,microphthalmia-associated transcription factor(MITF),tyrosinase(TYR)and tyrosinase-related protein 2(TYRP2)vs NC,which were 3.193±0.063,2.021±0.129,3.218±0.056,3.140±0.108 folds of the NC group respectively(P<0.01).Western blot analysis showed that the protein expression of MITF,TYR and TYRP2 in Alp-MC1 R group were 1.852±0.117,3.537±0.13 and 2.045±0.163 folds vs that in NC group respectively,with significant difference(P<0.01).In addition,Alp-MC1 R expression in sheep melanocytes caused a significant increase in melanin synthesis,which was 1.715±0.014 folds higher than that in the NC group(P<0.01).Our results suggested that Alp-MC1 R could be used to increase the expression of some hair color genes in sheep during melanogenesis and provided a potential application strategy for alpaca.