| Intramuscular fat content is closely related to tenderness,mouthfeel and juiciness of meat.Overdeposition of subcutaneous fat not only affects feed conversion efficiency,but also reduces carcass economic value.In this study,we sequenced the intramuscular and subcutaneous precursor fat cells in order to identify the key genes and lncRNAs related to the directional deposition of fat in meat ducks.The specific results of the experiment are as follows:1.There is significant difference between intramuscular and subcutaneous preadipocyte differentiationIn this experiment,intramuscular preadipocyte(IMP)and subcutaneous preadipocyte cells(SCP)were obtained by enzyme digestion method and differential adherence method.The precursor fat cells showed irregular spindle shape and long triangle shape,and there was no obvious difference between IMP and SCP shape.After induced differentiation,the shape of precursor fat cells became circular gradually.At the same time,lipid droplets gathered,indicating that the precursor fat cells differentiated into mature fat cells.The growth curve showed that the growth rate of IMP and SCP was "S type",which accorded with the growth law of primary cells.The growth rate of SCP was higher than that of IMP at the early stage of growth,and higher than that of SCP at the end of 6 days.The results of TG assay showed that IMP and SCP increased TG content,but TG content in SCP increased faster than that in IMP,and there was a significant difference in the 4th day of induction(P<0.05).The expression levels of PPARγ、FAS、FABP4 and LPL were detected by qRT-PCR.The results showed that SCP was higher than IMP,and the expression level of SCP was significantly higher than IMP in 4d and 6d.These results indicated that there was a significant change in the fat deposition of the two sites before and after induction,and a significant difference(P<0.05)after 4 days of induction.2.Differential Analysis of Intramuscular and Subcutaneous Adipocyte Differentiation in Total Transcriptional GroupIn this study,we screened differentially expressed genes and differentially expressed lncRNAs of intramuscular precursor fat cells IMP-0,intramuscular fat cells IMP-4,subcutaneous fat cells SCP-0 and subcutaneous fat cells SCP-4 by total transcription sequencing,and analyzed their functional enrichment.The study found that IMP-0-vs-IMP-4 had 1419 differentially expressed genes and 697 differentially expressed lncRNAs.SCP-0-vs-SCP-4 had 2307 differentially expressed genes and 1180 differentially expressed lncRNAs.IMP-0-vs-IMP-4 and SCP-0-vs-SCP-4 were plotted with Wayne diagrams to obtain the unique differentially expressed genes of each group.GR、PPARγ、FABP4、FABP5、ACSL1、APOA1、PLIN1、SGK1、FADS2 and the key differentially differentially genes,lncNURAD、110353236,MSTRG.11327.1、MSTRG.13937.5、MSTRG.1041.1、MSTRG.10027.2,MSTRG.10900.1、MSTRG.13938.2 and the key differentially differentially lncRNAs.3.Molecular Mechanism of lncNURAD Regulating Adipocyte DifferentiationThe whole transcriptome was screened to obtain the lIncNURAD-GR cis regulatory network.The expression trend of lncNURAD and GR in IMP was the same before and after induction by qRT-PCR,but opposite by SCP.By overexpression of lncNURAD,the expression of lncNURAD was 2.02 times higher than that of the control group,and When the lncNURAD was disturbed,the expression of shlncNURAD was decreased by 25 times than that of the control group shNC.Significant differences were found in the expression of key adipose genes after lncNURAD overexpression/interference.In conclusion,by comparing the difference of intramuscular and subcutaneous adipocytes and their differentiation,the molecular regulation mechanism of intramuscular and subcutaneous adipocytes can be detected,which provides a theoretical basis for further study on the metabolism and differentiation of adipose tissue in various parts of the body,and lays a foundation for improving meat quality of meat ducks and increasing economic benefits of enterprises. |
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