Identification of a novel microRNA important for melanogenesis in alpaca (Vicugna pacos).
Identification of a novel microRNA important for melanogenesis in alpaca (Vicugna pacos).
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DOI：
10.2527/jas.2014-8404
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发表时间：
2015-04
影响因子：
3.3
通讯作者：
S. Yang;R. Fan;Z. Shi;K. Ji;J. Zhang;H. Wang;M. Herrid;Q. Zhang;J. Yao;G. Smith;
中科院分区：
文献类型：
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作者：
S. Yang;R. Fan;Z. Shi;K. Ji;J. Zhang;H. Wang;M. Herrid;Q. Zhang;J. Yao;G. Smith;
动物毛色形成的分子机制还知之甚少。最近的研究表明，microRNA在哺乳动物的黑色素合成和毛色控制中发挥着重要作用。在之前的研究中，我们研究了棕色和白色羊驼皮肤中miRNA的表达谱，并发现了一种新的miRNA(命名为lpa-miR-nov-66)，它在白色皮肤中的表达显著高于棕色皮肤。本研究旨在确定这种新的miRNA在调节羊驼黑素细胞黑素合成中的功能作用。根据可溶性鸟苷环化酶(SGC)编码序列(CDS)中存在的结合位点，预测LPA-miR-NOV-66靶向sGC基因。在羊驼黑色素细胞中过表达lpa-miR-nov-66可在mRNA和蛋白水平上上调sGC的表达。在黑色素细胞中过表达lpa-miR-Nov-66还导致酪氨酸酶(TYR)、酪氨酸酶相关蛋白1(TYRP1)和小眼球转录因子(MITF)等关键黑素基因表达降低。我们的酶联免疫吸附试验显示，过表达lpa-miR-nov-66的黑素细胞环鸟苷一磷酸(CGMP)增加，环腺苷一磷酸(CAMP)减少。此外，过表达lpa-miR-nov-66也减少了培养的黑素细胞中黑色素的产生。结果支持lpa-miR-nov-66通过直接或间接靶向作用于黑素细胞，通过cAMP途径调节黑素合成。
The molecular mechanisms underlying the formation of coat colors in animals are poorly understood. Recent studies have demonstrated that microRNA play important roles in the control of melanogenesis and coat color in mammals. In a previous study, we characterized the miRNA expression profiles in alpaca skin with brown and white coat color and identified a novel miRNA (named lpa-miR-nov-66) that is expressed significantly higher in white skin compared to brown skin. The present study was conducted to determine the functional roles of this novel miRNA in the regulation of melanogenesis in alpaca melanocytes. lpa-miR-nov-66 is predicted to target the soluble guanylate cyclase (sGC) gene based on presence of a binding site in the sGC coding sequence (CDS). Overexpression of lpa-miR-nov-66 in alpaca melanocyes upregulated the expression of sGC both at the mRNA and protein level. Overexpression of lpa-miR-nov-66 in melanocyes also resulted in decreased expression of key melanogenic genes including tyrosinase (TYR), tyrosinase related protein 1 (TYRP1), and microphthalmia transcription factor (MITF). Our ELISA assays showed increased cyclic guanosine monophosphate (cGMP) but decreased cyclic adenosine monophosphate (cAMP) production in melanocytes overexpressing lpa-miR-nov-66. In addition, overexpression of lpa-miR-nov-66 also reduced melanin production in cultured melanocytes. Results support a role of lpa-miR-nov-66 in melanocytes by directly or indirectly targeting , which regulates melanogenesis via the cAMP pathway.