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This study performed a comprehensive expression profiling of genes expressed in the skin of goats with three different coat colors by Illumina Sequencing. A total of 91 significantly expressed genes were detected when comparing gray skin to white skin library and these included 74 up-regulated and 17 down-regulated genes in gray skin. There were 67 differentially expressed genes between brown skin and white skin libraries, 23 of which were up-regulated and 44 were down-regulated in brown skin. When we compared brown and gray libraries, 154 differentially expressed genes were found, of which 33 showed higher expression and 121 showed lower expression in brown skin. To our surprise, MC1R, MITF, TYR and KIT showed no significant difference in expression between the goats with three skin colors, whereas ASIP was detected in white skin but not in dark skins. In this study, PMEL, TRPM1, TYRP1 and DCT were significantly up-regulated in brown goat skin compare with gray and white skins. PMEL showed higher expression in gray goat skin compared with white goat skin, whereas there were no significant differences in the expression of TYRP1, TRPM1 and DCT between gray and white skin samples. In addition, ELOVL3 showed higher expression in gray goat skin than in brown and white skins, whereas there was no significant differences in the expression of ELOVL3 between brown and white skin samples. These results expand our understanding of the complex molecular mechanisms of skin physiology and melanogenesis in goat and provide a foundation for future studies.

Despite extensive research on the role of miR-135a in biological processes, very little attention has been paid to the regulation of its transcription. We have previously reported that miR-135a suppresses 3T3-L1 preadipocyte differentiation and adipogenesis by directly targeting the adenomatous polyposis coli (APC) gene and activating the canonical Wnt/β-catenin signaling pathway, but the regulatory elements that regulate the expression of the two isoforms of miR-135a (miR-135a-1 and miR-135a-2) remain poorly understood. Here, by using deletion analysis, we predicted two binding sites (-874/-856 and -2020/-2002) for the transcription factor Signal Transducers and Activators of Transcription 5a (STAT5a) within the core promoters of miR-135a-1 and miR-135a-2 (-1128/-556 and -2264/-1773), and the subsequent site-directed mutagenesis indicated that the two STAT5a binding sites regulated the activity of the miR-135a-1 and miR-135a-2 promoters. The binding of STAT5a to the miR-135a-1/2 core promoters in vitro and in cell culture was identified by electrophoretic mobility shift assays (EMSA) and chromatin immunoprecipitation (ChIP) assays. Overexpression and RNAi knockdown of STAT5a showed that the transcription factor regulated the endogenous miR-135a expression. Additionally, The expression time frame of STAT5a and APC indicated a potential negative feedback between them. In sum, the overall results from this study indicate that STAT5a regulates miR-135a transcription by binding to both miR-135a-1 and miR135a-2 promoter elements and the findings provide novel insights into the molecular regulatory mechanisms of miR-135a during adipogenesis.

Despite the rich variety in plumage color found in nature, genetic studies on how feather follicles affect pigmentation are often limited to animals that have black and white pigment. To test how gene expression influences plumage color, transcriptomes of chicken feather follicles with white, black, hemp, reed catkins, silvery grey, and landscape plumage colors were generated using Illumina sequencing. We generated six RNA-Seq libraries with over 25 million paired-end clean reads per library with percentage of paired-end clean reads ranging from 96.73 to 96.98%. 78% of the reads mapped to the chicken genome, and approximately 70% of the reads were mapped to exons and 6% mapped to introns. Transcriptomes of feather follicles producing hemp and land plumage were similar, but these two showed moderate differences compared with gray and reed colored plumage. The black and white follicle transcriptomes were most divergent from the other colors. We identified several candidate genes, including GPNMB, PMEL, TYRP1, GPR143, OCA2, SOX10, SLC45A2, KRT75, and TYR. All of these genes are known to induce pigment formation in mice. White feathers result from the lack of pigment formation, and our results suggest that the white chickens due to the recessive insertion mutation of TYR. The formation of black area size and color depth may be due to the expression levels of GPNMB, PMEL, TYRP1, GPR143, OCA2, SOX10, SLC45A2, KRT75, and TYR. The GO analysis of the differentially expressed genes (DEGs) revealed that DEGs in our transcriptome analysis were enriched in cytoskeleton and cell structure related pathways. The black plumage transcriptome showed significant differences in melanogenesis, tyrosine metabolism, and riboflavin metabolism compared with transcriptomes of other plumage colors. The transcriptome profiles of the different chicken plumage colors provide a valuable resource to understand how gene expression influences plumage color, and will be an important resource for identifying candidate genes in breeding programs.

Zinc finger protein 217 (Zfp217), a member of the kruppel-type zinc finger protein family, plays diverse roles in cell differentiation and development of mammals. Despite extensive research on the functions of Zfp217 in cancer, pluripotency and reprogramming, its physiological roles in adipogenesis remain unknown. Our previous RNA sequencing data suggest the involvement of Zfp217 in adipogenesis. In this study, the potential function of Zfp217 in adipogenesis was investigated through bioinformatics analysis and a series of experiments. The expression of Zfp217 was found to be gradually upregulated during the adipogenic differentiation in C3H10T1/2 cells, which was consistent with that of the adipogenic marker gene Pparg2. Furthermore, there was a positive, significant relationship between Zfp217 expression and adipocyte differentiation. It was also observed that Zfp217 could not only trigger proliferative defect in C3H10T1/2 cells, but also interact with Ezh2 and suppress the downstream target genes of Ezh2. Besides, three microRNAs (miR-503-5p, miR-135a-5p and miR-19a-3p) which target Zfp217 were found to suppress the process of adipogenesis. This is the first report showing that Zfp217 has the capacity to regulate adipogenesis.

MicroRNAs are endogenous, conserved, and non-coding small RNAs that function as post-transcriptional regulators of fat development and adipogenesis. Adipogenic marker genes, such as CCAAT/enhancer binding protein α (Cebpa), peroxisome proliferator-activated receptor γ (Pparg), adipocyte fatty acid binding protein (Ap2), and fatty acid synthase (Fas), are regarded as the essential transcriptional regulators of preadipocyte differentiation and lipid storage in mature adipocytes. Canonical Wnt/β-catenin signaling is recognized as a negative molecular switch during adipogenesis. In the present work we found that miR-135a-5p is markedly downregulated during the process of 3T3-L1 preadipocyte differentiation. Overexpression of miR-135a-5p impairs the expressions of adipogenic marker genes as well as lipid droplet accumulation and triglyceride content, indicating the importance of miR-135a-5p for adipogenic differentiation and adipogenesis. Further studies show that miR-135a-5p directly targets adenomatous polyposis coli (Apc), contributes to the translocation of β-catenin from cytoplasm to nucleus, and then activates the expressions of cyclin D1 (Ccnd1) and Cmyc, indicating the induction of canonical Wnt/β-catenin signaling. In addition, inhibition of APC with siRNA exhibits the same effects as overexpression of miR-135a-5p. Our findings demonstrate that miR-135a-5p suppresses 3T3-L1 preadipocyte differentiation and adipogenesis through the activation of canonical Wnt/β-catenin signaling by directly targeting Apc. Taken together, these results offer profound insights into the adipogenesis mechanism and the development of adipose tissue.

The words 'hair follicles' was replaced with 'feather follicles' in the title and the main text.

ASIP has been described to regulate lipid metabolism and acts as a paracrine factor in the regulation of adiposity. The RT-qPCR and Western Blotting was used to investigate the expression level of Agouti mRNA and Agouti signaling protein (ASIP) in viscera (heart, liver, spleen, lung and kidney) of goat with white, brown with gray and black coat color. The main trend of Agouti mRNA abundance represented white goat and liver tissue obviously. The protein was detected in all viscera tissues and it expressed white goat and kidney mainly. However, ASIP differential expresses in different viscera for goats weren't consistent with the mRNA levels. The differences might be the result from the transcription regulation, translation regulation, protein modified and protein interaction and so on.

Lysine-specific demethylase 1 (LSD1) functioned as a demethyl methylase gene, underlying a wide range of biological processes, including cancer, cell apoptosis, differentiation, and development. To further understand the functions of the porcine LSD1 gene, we first obtained cDNA sequence of porcine LSD1 gene, using in silico cloning method. We further found that the porcine LSD1 gene has two transcripts, in which cDNA sequences are 2,716 and 2,656 bp, ORF are 2,622 and 2,562 bp, respectively. Then, RT-PCR analysis showed that the LSD1 gene is expressed in various tissues and relatively higher in the tissues of ovary, kidney, and spleen. Besides, the LSD1 gene was expressed higher in the growth nonage and peaked at 3 days in muscle tissue. Meanwhile, the expression of two transcript variants of LSD1 gene presented the same change trend. Besides, the level of DNA methylation was approximately fourfold higher in a 3-day muscle than in an old pig (180 days), significantly positive related to the gene expression of LSD1 (R = 0.9362, P

Capra hircus, an economically important livestock, plays an indispensable role in the world animal fiber industry. To identify additional genes that may play important roles in coat color regulation, Illumina/Solexa high throughput sequencing technology was used to catalog the global gene expression profiles in the skin of three different coat colors goat (Lubei white goat (white), Jining gray goat (gray) and Jianyang big ear goat (brown)). The RNA-Seq analysis generated 83174342, 70222592 and 52091212 clean reads in white skin, gray skin and brown skin, respectively, which provided abundant data for further analysis. A total of 91 genes were differentially expressed between the gray skin and white skin libraries, with 74 upregulated and 17 genes downregulated. Between the brown skin and white skin libraries, there were 23 upregulated genes and 44 downregulated genes, while there were 33 upregulated genes and 121 downregulated genes between the brown skin and gray skin libraries. To our surprise, MC1R, MITF, TYR, KIT and KITLG showed no significant difference in the skin of three different coat colors and the expression of ASIP was only detected in white skin and not in gray and brown skins. The expression of PMEL,TRPM1, DCT, TYRP1 and ELOVL3 was validated by real-time quantitative polymerase chain reaction (qPCR) and the results of the qPCR were consistent with the RNA-seq except the expression of TYRP1 between the gray skin and white skin libraries. This study provides several candidate genes that may be associated with the development of diferent coat colors goat skin. More importantly, the fact that the ASIP gene was only detected in the white skin and not in the other dark skins and the MC1R gene showed no significant difference in expression between the three different coat colors goat is of particular interest for future studies that aim to elucidate theirs functional role in the regulation of skin color. These results will expand our understanding of the complex molecular mechanisms of skin physiology and melanogenesis in goat and provide a foundation for future studies.