During goat embryonic morphogenesis and postnatal initiation of hair follicle (HF) regeneration, dermal papilla (DP) cells play a vital role in hair formation. Growing evidence shows that microRNAs (miRNAs) participate in HF development and DP cell proliferation. However, the molecular mechanisms have not been thoroughly investigated. In this study, we utilized miRNA sequencing (miRNA-Seq) to identify differentially expressed miRNAs at different HF cycling stages (anagen and telogen). MiRNA-Seq has identified 411 annotated miRNAs and 130 novel miRNAs in which 29 miRNAs were up-regulated and 32 miRNAs were down-regulated in the anagen phase compared to the telogen phase. Target gene prediction and functional enrichment analysis indicated some major biological pathways related to hair cycling, such as Wnt signaling pathways, ECM-receptor interaction, VEGF signaling pathway, biosynthesis of amino acids, metabolic pathways, ribosome and oxidative phosphorylation. Also, we explored the function of chi-miR-30b-5p in regulating hair growth cycle. Similar to the HF cycling, DP cells were isolated from skin and used to investigate miRNA functions. The MTT and EdU assays showed that the viability and proliferation of DP cells were inhibited or promoted after the transfection of chi-miR-30b-5p mimic or inhibitor, respectively. Bioinformatics analysis revealed CaMKIIδ as a candidate target gene of chi-miR-30b-5p, and the dual-luciferase and western blot assay demonstrated that chi-miR-30b-5p bound to the 3’UTR of CaMKIIδ and further inhibited its translation. Chi-miR-30b-5p was found to be highly expressed in the telogen than that in the anagen phase and could inhibit the proliferation of DP cells by targeting CaMKIIδ. Our study provides new information on the regulatory functions of miRNAs during HF development.

中文翻译：

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Chi-miR-30b-5p通过靶向绒山羊山羊中的CaMKIIδ基因来抑制真皮乳头细胞增殖。

在山羊胚胎形态发生和毛囊（HF）再生的出生后启动过程中，真皮乳头（DP）细胞在毛发形成中起着至关重要的作用。越来越多的证据表明，microRNA（miRNA）参与了HF的发展和DP细胞的增殖。但是，分子机理尚未得到彻底研究。在这项研究中，我们利用miRNA测序（miRNA-Seq）来鉴定在不同的HF循环阶段（生长期和毛发生长期）中差异表达的miRNA。与端粒期相比，miRNA-Seq已鉴定出在生长期中有411个带注释的miRNA和130个新颖的miRNA，其中29个miRNA被上调，而32个miRNA被下调。靶基因预测和功能富集分析表明与毛发循环相关的一些主要生物学途径，例如Wnt信号传导途径，ECM-受体相互作用，VEGF信号传导途径，氨基酸的生物合成，代谢途径，核糖体和氧化磷酸化。此外，我们探索了chi-miR-30b-5p在调节头发生长周期中的功能。与HF循环相似，从皮肤中分离出DP细胞，并用于研究miRNA的功能。MTT和EdU分析表明，分别转染chi-miR-30b-5p模拟物或抑制剂后，DP细胞的活力和增殖受到抑制或促进。生物信息学分析显示，CaMKIIδ是chi-miR-30b-5p的候选靶基因，双重荧光素酶和Western blot分析表明，chi-miR-30b-5p与CaMKIIδ的3'UTR结合并进一步抑制了其翻译。发现Chi-miR-30b-5p在端粒中的表达高于生长期，并且可以通过靶向CaMKIIδ抑制DP细胞的增殖。我们的研究提供了有关HF开发过程中miRNA调控功能的新信息。