Screening and functional validation of lipid metabolism-related lncRNA-46546 based on the transcriptome analysis of early embryonic muscle tissue in chicken.

OBJECTIVE: The study was conducted to screen differentially expressed long noncoding RNA (lncRNA) in chickens by high-throughput sequencing and explore its mechanism of action on intramuscular fat deposition. METHODS: Herein, Rose crown and Cbb broiler chicken embryo breast and leg muscle lncRNA and mRNA expression profiles were constructed by RNA sequencing. A total of 96 and 42 differentially expressed lncRNAs were obtained in Rose crown vs Cobb broiler chicken breast and leg muscle, respectively. lncRNA-ENSGALT00000046546, with high interspecific variability and a potential regulatory role in lipid metabolism, and its predicted downstream target gene 1-acylglycerol-3-phosphate-O-acyltransferase 2 (AGPAT2), were selected for further study on the preadipocytes. RESULTS: lncRNA-46546 overexpression in chicken preadipocyte 2 cells significantly increased (p<0.01) the expression levels of AGPAT2 and its downstream genes diacylglycerol acyltransferase 1 and diacylglycerol acyltransferase 2 and those of the fat metabolism-related genes peroxisome proliferator-activated receptor γ, CCAAT/enhancer binding protein α, fatty acid synthase, sterol regulatory element-binding transcription factor 1, and fatty acid binding protein 4. The lipid droplet concentration was higher in the overexpression group than in the control cells, and the triglyceride content in cells and medium was also significantly increased (p<0.01). CONCLUSION: This study preliminarily concludes that lncRNA-46546 may promote intramuscular fat deposition in chickens, laying a foundation for the study of lncRNAs in chicken early embryonic development and fat deposition.

[Expression of ER, bcl-2, and p53 mRNA in early hybrid embryos of chicken-quail].

Using artificial insemination, 100 female quails were crossed with 10 male chickens. The eggs were collected and hatched in the same incubator. The sex of live hybrid embryos from 66 to 120 hatch hours was determined using multiply PCR of Wpkci. Total 300 male and female embryos at various hatch times were sampled and the relative mRNA abundance of ER, bcl-2, and p53 in the embryos was detected by RT-PCR using beta-actin as the internal standard. The effects of ER, bcl-2, and p53 on the early embryonic development for hybrids between chicken and quail were analyzed. The results showed ER mRNA expression of female hybrids were higher than male hybrids from 66 to 84 hatch hours with a highly significant difference (P<0.01), which indicated that the sex differentiation of hybrids was perhaps happened between 66 to 84 h of embryo stage. The obvious sequential expression of bcl-2 and p53 in the embryonic development indicated that the bcl-2 and p53 genes had an important effect on the development of the hybrid embryos.