Key Laboratory for Animal Genetics

Yangzhou, China

Key Laboratory for Animal Genetics

Yangzhou, China
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Xue Q.,Yangzhou University | Xue Q.,Key Laboratory for Animal Genetics | Zhang G.,Yangzhou University | Zhang G.,Key Laboratory for Animal Genetics | And 8 more authors.
PLoS ONE | Year: 2017

The early growth pattern, especially the age of peak growth, of broilers affects the time to market and slaughter weight, which in turn affect the profitability of the poultry industry. However, the underlying mechanisms regulating chicken growth and development have rarely been studied. This study aimed to identify candidate genes involved in chicken growth and investigated the potential regulatory mechanisms of early growth in chicken. RNA sequencing was applied to compare the transcriptomes of chicken muscle tissues at three developmental stages during early growth. In total, 978 differentially expressed genes (DEGs) (fold change ≥ 2; false discovery rate < 0.05) were detected by pairwise comparison. Functional analysis showed that the DEGs are mainly involved in the processes of cell growth, muscle development, and cellular activities (such as junction, migration, assembly, differentiation, and proliferation). Many of the DEGs are well known to be related to chicken growth, such as MYOD1, GH, IGF2BP2, IGFBP3, SMYD1, CEBPB, FGF2, and IGFBP5. KEGG pathway analysis identified that the DEGs were significantly enriched in five pathways (P < 0.1) related to growth and development: extracellular matrix-receptor interaction, focal adhesion, tight junction, insulin signaling pathway, and regulation of the actin cytoskeleton. A total of 42 DEGs assigned to these pathways are potential candidate genes inducing the difference in growth among the three developmental stages, such as MYH10, FGF2, FGF16, FN1, CFL2, MAPK9, IRS1, PHKA1, PHKB, and PHKG1. Thus, our study identified a series of genes and several pathways that may participate in the regulation of early growth in chicken. These results should serve as an important resource revealing the molecular basis of chicken growth and development. © 2017 Xue et al. This is an open access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.


Zhang Y.,Yangzhou University | Zhang Y.,Key Laboratory for Animal Genetics | Wang Y.,Yangzhou University | Wang Y.,Key Laboratory for Animal Genetics | And 20 more authors.
PLoS ONE | Year: 2017

An efficient genome editing approach had been established to construct the stable transgenic cell lines in the domestic chicken (Gallus gallus domesticus) at present. Our objectives were to investigate gene function in the differentiation process of chicken embryonic stem cells (ESCs) into spermatogonial stem cells(SSCs). Three guides RNA (gRNAs) were designed to knockout the Stra8 gene, and knockout efficiency was evaluated in domestic chicken cells using cleavage activity of in vitro transcription of gRNA, Luciferase-SSA assay, T7 endonuclease I assay(T7E1) and TA clone sequence. In addition, the Cas9/ gRNA plasmid was transfected into ESCs to confirm the function of Stra8. SSA assay results showed that luciferase activity of the vector expressing gRNA-1 and gRNA- 2 was higher than that of gRNA-3. TA clone sequencing showed that the knockdown efficiency was 25% (10/40) in DF-1 cells, the knockdown efficiency was 23% (9/40) in chicken ESCs. T7E1 assay indicated that there were cleavage activity for three individuals, and the knockdown efficiency was 12% (3/25). Cell morphology, qRT-PCR, immunostaining and FCS indicated that Cas9/gRNA not only resulted in the knockout of Stra8 gene, but also suggested that the generation of SSCs was blocked by the Stra8 gene knockdown in vitro. Taken together, our results indicate that the CRISPR/Cas9 system could mediate stable Stra8 gene knockdown in domestic chicken's cells and inhibit ECSs differentiation into SSCs. Copyright: © 2017 Zhang et al. This is an open access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.


Zhang T.,Yangzhou University | Zhang T.,Key Laboratory for Animal Genetics | Zhang X.,Yangzhou University | Zhang X.,Key Laboratory for Animal Genetics | And 11 more authors.
PLoS ONE | Year: 2017

Long noncoding RNAs (lncRNAs) regulate metabolic tissue development and function, including adipogenesis. However, little is known about the function and profile of lncRNAs in intramuscular preadipocyte differentiation in chicken. Here, we identified lncRNAs in chicken intramuscular preadipocytes at different differentiation stages using RNA sequencing. A total of 1,311,382,604 clean reads and 25,435 lncRNAs were obtained from 12 samples. In total, 7,433 differentially expressed genes (4,698 lncRNAs and 2,735 mRNAs) were identified by pairwise comparison. These 7,433 differentially expressed genes were grouped into 11 clusters based on their expression patterns by K-means clustering. Using Weighted Gene Coexpression Network Analysis, we identified four stage-specific modules positively related to I0, I2, I4, and I6 stages and two stage-specific modules negatively related to I0 and I2 stages, respectively. Many well-known and novel pathways associated with intramuscular preadipocyte differentiation were identified. We also identified hub genes in each stage-specific module and visualized them in Cytoscape. Our analysis revealed many highly-connected genes, including XLOC-058593, BMP3, MYOD1, and LAMP3. This study provides a valuable resource for chicken lncRNA study and improves our understanding of the biology of preadipocyte differentiation in chicken. © 2017 Zhang et al. This is an open access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.


Wang B.,Yangzhou University | Wang B.,Key Laboratory for Animal Genetics | Pang M.,Jiangsu Academy of Agricultural Sciences | Xie X.,Jiangsu Academy of Agricultural Sciences | And 19 more authors.
Food Analytical Methods | Year: 2017

This manuscript describes a rapid, sensitive, and specific approach that utilizes ultra-performance liquid chromatography-electrospray ionization tandem triple quadrupole mass spectrometry (UPLC-ESI/MS/MS) for the selective determination of amoxicillin, amoxicillin metabolites, and ampicillin in chicken tissue samples (muscle, liver, and kidney). This optimized method was validated according to the requirements defined by the European Union and the Food and Drug Administration. The extract obtained after extraction and purification of chicken tissues was injected into a UPLC system coupled to a mass spectrometer operated in the positive electrospray MS/MS mode for analysis. The chicken tissues were spiked with the target compounds at levels of 25.0, 50.0, and 100.0 μg/kg, and the results showed that the extraction recoveries were all higher than 84% and the relative standard deviations did not exceed 20%. The limits of detection and limits of quantification were 0.01–1.36 and 0.05–5.44 μg/kg, respectively. The decision limits were 52.62–57.26 μg/kg, and the detection capabilities were 55.23–64.51 μg/kg. Finally, the new approach was verified through quantitative determination of the analytes in tissues from 30 commercial chickens obtained from local supermarkets. [Figure not available: see fulltext.] © 2017 Springer Science+Business Media New York


Wei Y.,Yangzhou University | Wei Y.,Key Laboratory for Animal Genetics | Wei Y.,Jiangxi Academy of Agricultural science | Zhang G.X.,Yangzhou University | And 11 more authors.
Genetics and Molecular Research | Year: 2016

The growth trait is important in poultry production. We analyzed the association between single nucleotide polymorphisms (SNPs) in the Myf5 and MyoG gene and Bian chicken growth traits. SNPs in candidate genes of the Bian chickens were detected by the polymerase chain reaction-single strand conformation polymorphism method. Two mutation loci and six genotypes were identified in each candidate gene. In terms of growth traits, least square analysis showed that the FF genotype of the MyoG was the advantage genotype and the IJ genotype of the Myf5 was the disadvantage genotype for growth trait in Bian chicken. Correlation analysis suggested that the different combination genotypes between Myf5 and MyoG genes had a significant effect on growth traits in Bian chickens. The result suggested that MyoG and Myf5 genes can be used in marker-assisted selection for improving the growth trait in Bian chicken. © 2016 The Authors.


Zhang Y.,Yangzhou University | Zhang Y.,Key Laboratory for Animal Genetics | Wang Y.,Yangzhou University | Wang Y.,Key Laboratory for Animal Genetics | And 10 more authors.
PLoS ONE | Year: 2016

Several inducers have been used to differentiate embryonic stem cells (ESCs) into male germ cells but the induction process has been inefficient. To solve the problem of low efficiency of inducer for ESCs differentiation into male germ cells, all-trans retinoic acid (ATRA), Am80(the retinoic acid receptor agonist), and estradiol (E2) was used to induce ESCs to differentiate into male germ cells in vitro. ESCs were cultured in media containing ATRA, Am80, or E2 respectively which can differentiate ESCs into a germ cell lineage. In process of ATRA and Am80 induction Group, germ cell-like cells can be observed in 10 days; but have no in E2 induction Group. The marker genes of germ cell: Dazl, Stra8, C-kit, Cvh, integrinα6, and integrinβ1 all showed a significant up-regulation in the expression level. The ATRA-induction group showed high expression of C-kit and Cvh around 4 days, and integrinα6 and integrinβ1 were activated on day 10, respectively, while the E2-,Am80-induction group showed a high expression of C-kit as early as 4 days immunocytochemistry results shown that, integrinα6 and integrinβ1 could be detected in the ATRA-, Am80-, and E2-induction group, Positive clones in the ATRA group were greater in number than those in the other two groups. we conclued that ATRA, Am80, and E2 can promote the expression of the corresponding genes of germ cells, and had different effect on the differentiation of ESCs into male germ cells. ATRA was the most effective inducer of germ cell differentiation. © 2016 Zhang et al. This is an open access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.


Tang Y.,Yangzhou University | Tang Y.,Key Laboratory for Animal Genetics | Zhang T.,Yangzhou University | Zhang T.,Key Laboratory for Animal Genetics | And 13 more authors.
Molecular Biology Reports | Year: 2014

The objective of this study was to analyze possible associations between single nucleotide polymorphisms (SNPs) in the Myf5 gene with chicken growth and reproductive traits. SNPs in Myf5 of the Jinghai yellow chicken were detected by the polymerase chain reaction single-strand conformation polymorphism method and the haplotypes were analyzed. Eight SNPs were identified in the exons of Myf5. Nine haplotypes were established in a group of 379 Jinghai yellow chickens. In terms of growth traits, least square analysis showed that haplotype H1H5 had significant effects on weight at weeks 8 and 12 (P < 0.05). Haplotype H2H6 had significant effects on weight at weeks 12 and 14 (P < 0.05). For reproductive traits, H1H5 had higher body weight for the first egg than H1H4 and H2H4 (P < 0.05), and H1H3 (P < 0.01). H1H3 had a poor performance in average egg weight at 300 days. On the other hand, H1H3 had an advantage in egg number at 300 days. The results showed that SNPs of Myf5 have certain effects on growth and reproductive traits in Jinghai yellow chickens, which can be used in marker-assisted selection to accelerate chicken genetic progress. © 2014, Springer Science+Business Media Dordrecht.

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