College of Animal Science & Veterinary Medicine, Shenyang Agricultural University, Shenyang, P. R. China.
Anim Biotechnol. 2024 Nov;35(1):2356110. doi: 10.1080/10495398.2024.2356110. Epub 2024 May 28.
The inducing activation event of secondary hair follicle (SHF)-stem cells is considered a key biological process in the SHF regeneration, and the morphogenesis of cashmere fiber in cashmere goats. The miR-361-5p was essentially implicated in the induced activation of SHF-stem cells of cashmere goats, but its functional mechanisms are unclear. Here, we confirmed miR-361-5p was significantly downregulated in anagen SHF bugle of cashmere goats compared with that at telogen, and miR-361-5p expression was significantly lower in SHF-stem cells after activation than its counterpart before activation. Further, we found that miR-361-5p could negatively regulate the induced activation event of SHF-stem cells in cashmere goats. Mechanistically, through dual-luciferase reporter assays, miR-361-5p specifically bound with mRNA in SHF-stem cells of cashmere goats and negatively regulated the expression of gene. Also, through overexpression/knockdown analysis of gene, our results indicated that upregulated the expression of Wnt/β-catenin pathway related genes in SHF-stem cells. Moreover, based on TOP/FOP-flash Wnt report assays, the knockdown of miR-361-5p promotes the Wnt/β-catenin pathway activation through upregulating the expression in SHF-stem cells. Finally, we demonstrated that miR-361-5p negatively regulated the induced activation of SHF-stem cells through mediated Wnt/β-catenin pathway in cashmere goats.
次生毛囊(SHF)-干细胞的诱导激活事件被认为是 SHF 再生和绒山羊绒纤维形态发生的关键生物学过程。miR-361-5p 在内质网应激诱导的 SHF-干细胞激活中起关键作用,但它的功能机制尚不清楚。在这里,我们证实 miR-361-5p 在绒山羊生长期 SHF 喇叭中显著下调,与休止期相比,miR-361-5p 在激活后的 SHF-干细胞中表达明显低于激活前。此外,我们发现 miR-361-5p 可以负调控绒山羊 SHF-干细胞的诱导激活事件。通过双荧光素酶报告基因实验,我们发现 miR-361-5p 可以与绒山羊 SHF-干细胞中的 mRNA 特异性结合,并负调控 基因的表达。此外,通过过表达/敲低 基因的分析,我们的结果表明, 基因上调了 SHF-干细胞中 Wnt/β-catenin 通路相关基因的表达。此外,基于 TOP/FOP-flash Wnt 报告基因实验,miR-361-5p 的敲低通过上调 SHF-干细胞中 基因的表达促进了 Wnt/β-catenin 通路的激活。最后,我们证明 miR-361-5p 通过 基因介导的 Wnt/β-catenin 通路负调控绒山羊 SHF-干细胞的诱导激活。