Transcriptome sequencing analysis of the role of miR-499-5p and SOX6 in chicken skeletal myofiber specification.

MicroRNAs (miRNAs) might play critical roles in skeletal myofiber specification. In a previous study, we found that chicken miR-499-5p is specifically expressed in slow-twitch muscle and that its potential target gene is SOX6. In this study, we performed RNA sequencing to investigate the effects of SOX6 and miR-499-5p on the modulation and regulation of chicken muscle fiber type and its regulatory mechanism. The expression levels of miR-499-5p and SOX6 demonstrated opposing trends in different skeletal muscles and were associated with muscle fiber type composition. Differential expression analysis revealed that miR-499-5p overexpression led to significant changes in the expression of 297 genes in chicken primary myoblasts (CPMs). Myofiber type-related genes, including MYH7B and CSRP3, showed expression patterns similar to those in slow-twitch muscle. According to functional enrichment analysis, differentially expressed genes were mostly associated with muscle development and muscle fiber-related processes. SOX6 was identified as the target gene of miR-499-5p in CPM using target gene mining and luciferase reporter assays. SOX6 knockdown resulted in upregulation of the slow myosin genes and downregulation of fast myosin genes. Furthermore, protein-protein interaction network analysis revealed that MYH7B and RUNX2 may be the direct targets of SOX6. These results indicated that chicken miR-499-5p may promote slow-twitch muscle fiber formation by repressing SOX6 expression. Our study provides a dataset that can be used as a reference for animal meat quality and human muscle disease studies.

A gene co-expression network analysis of the candidate genes and molecular pathways associated with feather follicle traits of chicken skin.

Back and thigh skin of chickens showed significant differences in the thickness and the feather follicle density and size, which are important traits for slaughtered chickens' appearance. In the present study, global gene expression profiling was conducted in the back and thigh skin of chickens using Microarray technology. The results showed that 676 genes were differentially expressed between back and thigh skin. The expression of the differentially expressed genes (DEGs), including PPP1R3C, IGF1, PTCHD1, HOXB6, FGF9, CAMK4, SHH, BMP8B, FOXN1 and PTGER2, was validated by real-time quantitative polymerase chain reaction (RT-qPCR), and the results were consistent with microarray results. Functional analysis revealed that the DEGs were significantly involved in cell proliferation, differentiation, apoptosis, adhesion and transport process, and the pathways were significantly mapped into the ECM-receptor interaction, peroxisome, focal adhesion, Hedgehog and PPAR signalling pathways. Protein-protein interaction network analysis suggested that signalling pathways related to feathers morphogenesis and development, such as Wnt, FGF, MAPK, SHH and BMP signalling pathways, occupied important positions in the network. Genes involved in these signalling pathways and adhesion molecules might play a vital role in skin and feather follicle development. Further single nucleotide polymorphism (SNP) association analysis of Wnt3A showed that the AC genotype of SNP g.255361 C>A significantly increased the feather follicle density of thigh skin. Our findings may provide new insights on candidate genes and pathways related to skin and feather follicle formation of chickens.

[Molecular genetic diversity of Fujian domestic duck breeds].

By using 28 micro-satellite markers with better polymorphism, this paper studied the genetic diversity of four Fujian provincial domestic duck breeds Jinding, Putian black, Liancheng white, and Shanma. According to the alleles frequencies, the polymorphic information content, average heterozygosity, anaqular genetic distance (DA) and Nei' s standard genetic distance (DS) for each breed were calculated. Based on DA and DS, four dendrograms were obtained by neighbor-joining (NJ) and UPGMA methods. The results showed that the average heterozygosity of the four duck breeds was 0. 5353, indicating that the protection of the genetic diversity of these breeds should be strengthened. The orders of the two types of genetic distances among the breeds were accordant, and the dendrograms were the same, reflecting that much more micro-satellite loci should be adopted to obtain more universal conclusions when the genetic diversity was analyzed. The phylogenetic relationships among the four duck breeds were in accordance with their economic types and ecological localities.

[Analysis of genetic diversity of Sichuan indigenous chicken breeds using microsatellite markers].

Thirty microsatellite markers with medium or high polymorphisms were selected to detect the genetic diversity of 8 indigenous chicken breeds in Sichuan. According to the allele frequencies of 30 microsatellite sites, mean heterozygosity (H), polymorphism information content (PIC) and DA genetic distances were calculated for each breeds. The results showed that 24 of 30 microsatellite sites were highly polymorphic, so the 24 microsatellite markers were effective markers for analysis of genetic relationship among chicken breeds. The mean heterozygosity of 8 chicken breeds was all over 0.5. The highest was the Luning chicken (0.681), and the lowest was the Jiuyuan Dark chicken. The high diversity of 8 chicken breeds might be caused by the traffic obstruction(geographic isolation). The results of the heterozygosity were consistent with that of PIC. UPGMA tree was completed through analysis of DA genetic distances. Emei Dark chicken, Miyi chicken, Luning chicken and Jiuyuan Dark chicken were the first group: Miyi chicken and Luning chicken were grouped firstly, then Emei Dark chicken were grouped with them in shorter time distances, and Jiuyuan Dark chicken were grouped with them at last. Shimiancao Ke chicken Xingwen Silky chicken and Muchuan Silky were the second group: Xingwen Silky chicken and Muchuan Silky were grouped firstly, and then Shimiancao Ke chicken was grouped with them. Liangshangya Ying chicken had its own branch. The result of UPGM was consistent with the genesis, breeding history, differentiation and location of 8 chicken breeds.