Identification of a Novel miRNA in Melanocytes of Alpaca (Lama Pacos)

• 作 者: RuiwenFan[1]ShanshanYang[1]ZhanquanShi[1]KaiyuanJi[1]XiaoyunJia[1]JianboYao[2]GeorgeWSmith[3]ChangshengDong[1]
• 机 构: College of Animal Science and Technology, Shanxi Agricultural University, Taigu 030801, China
• 出 处: 中国畜牧兽医学会动物解剖及组织胚胎学分会第十八次学术研讨会
• 发表日期: 2014年期

　　From the previous deep sequencing of alpaca skins, one of novel miRNAs expressed differently in white and black alpaca skin (named alp-miR066).According to the target gene prediction, one of the target genes is soluble guanylate cyclase (sGC).To get the function of alp-miR-066 in alpaca skin, it was over expressed in alpaca melanocytes in vitro.The expression of miRNA-066 was tested by real-time PCR and the results showed it expressed increased.On the pathway of cAMP in melanocytes, cGMP was increased by ELASA assay, while cAMP was decreased.The expression of sGC, TYR, TYRP1, TYRP2 and MITF protein were tested by western blotting, and the results showed that sGC was increased and TYR, TYRP1 and MITF protein were decreased.Moreover, the number of melanogenic melanocytes was less and the amount of melanins production was 0.7 fold in alp-miR066 transfection group, compared to Negative control(NC) transfection group.All those results suggested that alp-miR-066 might regulate melanogenesis on the pathway of cAMP in alpaca melanocytes by targeting sGC.