Dataset Information

Inner Mongolian Cashmere Goat Secondary Follicle Development Regulation Research Based on mRNA-miRNA Co-analysis.

ABSTRACT: Inner Mongolia cashmere goats, as an important part of animal husbandry production, play an important role in animal fiber industry. In recent years, scientific research has made a lot of explorations on the molecular regulation mechanism of hair follicle cycle growth, but few studies have been reported on the development of cashmere hair in fetal period. This study was based on the completion of 21 skin samples of mRNA and miRNA sequencing in 7 fetal periods (45 days, 55 days,65 days,75 days,95 days,115 days and 135 days) of the Inner Mongolia Cashmere goat. The target genes of miRNA associated with the development of secondary hair follicles in the cashmere goats were selected through the combination analysis of mRNA and miRNA data. Then the overexpression vector was constructed and the interaction between the miRNA and the target gene was identified by Dual-Luciferase Reporter Gene System. The function and interaction relationship of chi-miR-199a-5p and TGF-?2 were verified by RT-qPCR and western blot at the level of the fibroblasts in Inner Mongolia Cashmere goat. It provides a theoretical basis for further study of miRNA and its target genes regulating the occurrence and development of skin hair follicles. As the result shows, the expression trends of 7 genes (BAMBI, SMAD1, LTBP1, PPP2R1B, ID4, BMP8B and PITX2) and 7 miRNA (chi-miR-17-5p, chi-miR-125b-3p, chi-miR-21-5p, chi-miR-143-5p and chi-miR-106b-5p) in the skin samples for the seven stages of the fetus were shown to be consistent with the sequencing results. the results of sequencing are reliable. The correlation coefficient of TGF-?2 and chi-miR-199a-5p in fetal 45d-135d expression is -0.84, showing a strong negative correlation, The target relationship was preliminarily judged. The results of double luciferase vector report showed that chi-miR-199a-5p significantly decreased the expression of luciferase in TGF-?2 3'UTR, It is determined that there is a reciprocal relationship between them at a specific time. We transfected chi-miR199a-5p-FAM mimics into fibroblasts cultured in vitro from Inner Mongolia cashmere goats. After transfection, the cells were harvested to extract total RNA and protein. The mRNA and protein expression levels of TGF-?2 in fibroblasts were detected by RT-qPCR and western blot. It was verified that chi-miR-199a-5p inhibited TGF-?2 expression at both mRNA and protein translation levels in fibroblasts. At the same time, it was again proved that the TGF-?2 gene is a target gene of chi-miR199a-5p.

SUBMITTER: Han W

PROVIDER: S-EPMC7066195 | biostudies-literature | 2020 Mar

REPOSITORIES: biostudies-literature

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Inner Mongolian Cashmere Goat Secondary Follicle Development Regulation Research Based on mRNA-miRNA Co-analysis.

Han Wenjing W Yang Feng F Wu Zhihong Z Guo Fuqiang F Zhang Junjie J Hai Erhan E Shang Fangzheng F Su Rui R Wang Ruijun R Wang Zhiying Z Liu Zhihong Z Zhao Yanhong Y Wang Zhixin Z Zhang Yanjun Y Li Jinquan J

Scientific reports 20200311 1

Inner Mongolia cashmere goats, as an important part of animal husbandry production, play an important role in animal fiber industry. In recent years, scientific research has made a lot of explorations on the molecular regulation mechanism of hair follicle cycle growth, but few studies have been reported on the development of cashmere hair in fetal period. This study was based on the completion of 21 skin samples of mRNA and miRNA sequencing in 7 fetal periods (45 days, 55 days,65 days,75 days,95 ...[more]

PMID: 32161290

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Project description:Inner Mongolian Cashmere goat is an excellent local breed selected for the dual-purpose of cashmere and meat. There are three lines of Inner Mongolian Cashmere goat: Erlangshan, Alashan and Aerbasi. Cashmere is a kind of precious textile raw material with a high price. Cashmere is derived from secondary hair follicle (SHF), while hair is derived from primary hair follicle (PHF). The growth cycle of SHF of cashmere goat is 1 year, and it can be divided into three different stages: anagen, catagen and telogen. In this study, we tried to find some important influence factors of SHF growth cycle in skin tissue from Inner Mongolian Cashmere goats by RNA sequencing (RNA-Seq). Three female Aerbasi Inner Mongolian Cashmere goats (2 years old) were used as experimental samples in this study. Skin samples were collected in September (anagen), December (catagen) and March (telogen) at dorsal side from cashmere goats. Results showed that over 511 396 044 raw reads and 487 729 890 clean reads were obtained from sequence data. In total, 51 different expression genes (DEGs) including 29 downregulated genes and 22 upregulated genes were enriched in anagen-catagen comparing group. The 443 DEGs contained 117 downregulated genes and 326 upregulated genes that were enriched in catagen-telogen comparing group. In telogen-anagen comparing group, 779 DEGs were enriched including 582 downregulated genes and 197 upregulated genes. The result of gene ontology (GO) annotation showed that DEGs are in different growth cycle periods, and enriched GO items are mostly related to the transformation of cell and protein. The Kyoto Encyclopedia of Genes and Genomes (KEGG) enrichment result indicated that metabolic process has a great impact on SHF growth cycle. Based on the results of a comprehensive analysis of differentially expressed genes, GO enrichment and KEGG enrichment, we found that FGF5, FGFR1 and RRAS had an effect on the hair follicle growth cycle. The results of this study may provide a theoretical basis for further research on the growth and development of SHF in Inner Mongolian Cashmere goats.

Project description:BackgroundInner Mongolian cashmere goats have hair of excellent quality and high economic value, and the skin hair follicle traits of cashmere goats have a direct and important effect on cashmere yield and quality. Circular RNA has been studied in a variety of tissues and cells.ResultIn this study, high-throughput sequencing was used to obtain the expression profiles of circular RNA (circRNA) in the hair follicles of Inner Mongolian cashmere goats at different embryonic stages (45, 55, 65, and 75 days). A total of 21,784 circRNAs were identified. At the same time, the differentially expressed circRNA in the six comparison groups formed in the four stages were: d75vsd45, 59 upregulated and 33 downregulated DE circRNAs; d75vsd55, 61 upregulated and 102 downregulated DE circRNAs; d75vsd65, 32 upregulated and 33 downregulated DE circRNAs; d65vsd55, 67 upregulated and 169 downregulated DE circRNAs; d65vsd45, 96 upregulated and 63 downregulated DE circRNAs; and d55vsd45, 76 upregulated and 42 downregulated DE circRNAs. Six DE circRNA were randomly selected to verify the reliability of the sequencing results by quantitative RT-PCR. Subsequently, the circRNA corresponding host genes were analyzed by the Gene Ontology (GO) and the Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway. The results showed that the biological processes related to hair follicle growth and development enriched by GO mainly included hair follicle morphogenesis and cell development, and the signaling pathways related to hair follicle development included the Notch signaling pathway and NF-κB signaling pathway. We combined the DE circRNA of d75vsd45 with miRNA and mRNA databases (unpublished) to construct the regulatory network of circRNA-miRNA-mRNA, and formed a total of 102 pairs of circRNA-miRNA and 126 pairs of miRNA-mRNA interactions. The binding relationship of circRNA3236-chi-miR-27b-3p and circRNA3236-chi-miR-16b-3p was further verified by dual-luciferase reporter assays, and the results showed that circRNA3236 and chi-miR-27b-3p, and circRNA3236 and chi-miR-16b-3p have a targeted binding relationship.ConclusionTo summarize, we established the expression profiling of circRNA in the fetal skin hair follicles of cashmere goats, and found that the host gene of circRNA may be involved in the development of hair follicles of cashmere goats. The regulatory network of circRNA-miRNA-mRNA was constructed and preliminarily verified using DE circRNAs.

Project description:Cashmere goat from Inner Mongolia is an excellent local breed in China, and the related cashmere product is a kind of precious textile raw material with high price. Cashmere is generated from secondary hair follicles, which has obvious annual periodicity and includes three different stages: anagen, catagen, and telogen. Therefore, we investigated skin transcriptome data for 12 months using weighted gene co-expression network analysis (WGCNA) to explore essential modules, pathways, and genes responsible for the periodic growth and development of secondary hair follicles. A total of 17 co-expression modules were discovered by WGCNA, and there is a strong correlation between steelblue module and month (0.65, p = 3E-09), anagen (0.52, p = 1E-05), telogen (-0.6, p = 8E-08). Gene expression was generally high during late anagen to catagen (June to December), while expression was downregulated from telogen to early anagen (January-May), which is similar to the growth rule of hair follicle cycle. KEGG pathway enrichment analyses of the genes of steelblue module indicated that genes are mainly enriched in Cell cycle, Wnt signaling pathway, p53 signaling pathway and other important signal pathways. These genes were also significantly enriched in GO functional annotation of the cell cycle, microtubule movement, microtubule binding, tubulin binding, and so on. Ten genes (WIF1, WNT11, BAMBI, FZD10, NKD1, LEF1, CCND3, E2F3, CDC6, and CDC25A) were selected from these modules, and further identified as candidate biomarkers to regulate periodic development of hair follicles using qRT-PCR. The Wnt signaling pathway and Cell cycle play an important role in the periodic development of hair follicles. Ten genes were identified as essential functional molecules related to periodic development of hair follicle. These findings laid a foundation for understanding molecular mechanisms in biological functions such as hair follicle development and hair growth in cashmere goats.

Dataset Information

Inner Mongolian Cashmere Goat Secondary Follicle Development Regulation Research Based on mRNA-miRNA Co-analysis.

Publications

Inner Mongolian Cashmere Goat Secondary Follicle Development Regulation Research Based on mRNA-miRNA Co-analysis.

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