College of Animal Science, Inner Mongolia Agricultural University, Hohhot, Inner Mongolia, 010018, China.
College of Veterinary Medicine, Inner Mongolia Agricultural University, Hohhot, Inner Mongolia, 010018, China.
Funct Integr Genomics. 2022 Oct;22(5):835-848. doi: 10.1007/s10142-022-00842-y. Epub 2022 Apr 29.
microRNA (miRNA) is a type of endogenous short-chain non-coding RNA with regulatory function found in eukaryotes, which is involved in the regulation of a variety of cellular and biological processes. However, the research on the development of cashmere goat secondary hair follicles is still relatively scarce. In this study, small RNA libraries and mRNA libraries of 45 days, 55 days, 65 days, and 75 days of fetal skin of cashmere goats were constructed, and the constructed libraries were sequenced using Illumina Hiseq4000, and the expression profiles of miRNA and mRNA in cashmere goat fetal skin were obtained. The differentially expressed miRNAs and mRNAs in six control groups were identified and the qRT-PCR experiment shows that the sequencing results are accurate. Sixty-six miRNAs related to secondary hair follicle development were screened, and used TargetScan and miRanda to predict 33 highly expressed miRNA target genes. At the same time, 664 mRNAs related to the development of secondary hair follicles were screened, and GO enrichment and KEGG pathway analysis were performed. It was found that some miRNA target genes were consistent with the screening results of mRNAs related to secondary hair follicle development and were enriched in Notch signaling pathway, TGF-β signaling pathway. Therefore, miR-145-5p-DLL4, miR-27b-3p-DLL4, miR-30e-5p-DLL4, miR-193b-3p-TGF-β1, miR-181b-5p-NOTCH2, and miR-103-3p-NOTCH2 regulatory network related to the development of secondary hair follicles were constructed and the results of dual-luciferase reporter gene assay indicated that there is a targeted relationship between chi-miR-30e-5p and DLL4, which will provide a basis for molecular mechanism of miRNA-mRNA in the development of the hair follicles in cashmere goats.
微小 RNA(miRNA)是一种在真核生物中发现的具有调节功能的内源性短链非编码 RNA,参与调节多种细胞和生物过程。然而,有关绒山羊次级毛囊发育的研究仍然相对较少。本研究构建了 45、55、65 和 75 日龄绒山羊胎儿皮肤的小 RNA 文库和 mRNA 文库,使用 Illumina Hiseq4000 对构建的文库进行测序,获得了绒山羊胎儿皮肤中 miRNA 和 mRNA 的表达谱。鉴定了 6 个对照组中差异表达的 miRNA 和 mRNA,并通过 qRT-PCR 实验表明测序结果准确。筛选出 66 个与次级毛囊发育相关的 miRNA,使用 TargetScan 和 miRanda 预测了 33 个高表达 miRNA 的靶基因。同时,筛选出 664 个与次级毛囊发育相关的 mRNA,并进行了 GO 富集和 KEGG 通路分析。结果发现,一些 miRNA 靶基因与次级毛囊发育相关的 mRNA 筛选结果一致,并富集在 Notch 信号通路、TGF-β 信号通路中。因此,构建了 miR-145-5p-DLL4、miR-27b-3p-DLL4、miR-30e-5p-DLL4、miR-193b-3p-TGF-β1、miR-181b-5p-NOTCH2 和 miR-103-3p-NOTCH2 等与次级毛囊发育相关的 miRNA-mRNA 调控网络,双荧光素酶报告基因检测结果表明 chi-miR-30e-5p 与 DLL4 存在靶向关系,为绒山羊毛囊发育过程中 miRNA-mRNA 的分子机制提供了依据。