Expression And Function Of Lnc₀02783 In Goat Skeletal Muscle

In this experiment,Lnc₀02783 was selected as the research object by bioinformatics and EdU cell proliferation test.Then,by analyzing the expression profiles and functions of Lnc₀02783 and its predicted target genes in skeletal muscle tissues and cells of Anhui white goats,it can provide a theoretical basis for goat breeding at the molecular level.（1）Goat skeletal muscle satellite cells（SCs）were isolated by two-step enzymatic digestion with type I collagenase and trypsin.In addition,based on the muscle sequencing data of the previous Anhui white goats in the research group,miRanda,PITA and RNAhybrid were used to predict the miRNAs interacting with the differential LncRNA,and 8 LncRNAs（Lnc₀02506,Lnc₀14160,Lnc₀02488,Lnc₀08909,Lnc₀08726,Lnc₀02783,LNC₀01690,LNC-009639）and 6 miRNAs（chi-miR-214-3p,chi-miR-193b-5p,chi-let-7a-5p,chi-miR-188-5p,chi-let-7b）-3p,chi-miR-124a)were selected as a preliminary study object to carried out the next experiment.（2）After interference with 8 LncRNAs and overexpression of 6 miRNAs,the results of the EdU cell proliferation assay showed that Lnc₀02783 and its predicted target gene miR-188-5p promoted and inhibited cell proliferation,respectively.（3）Collecting tissue samples of Anhui white goats,including the longissimus dorsi muscle tissue of 6 different developmental stages of F65,F90,F120,F135,B1 and B90;heart,liver,spleen,lung and kidney,the longissimus dorsi muscle and leg muscles of adult Anhui white goat.At the same time,goat SCs samples were collected for cultivate 2 d,3 d and 4 d,and the expression of Lnc₀02783 and miR-188-5p in goat tissues and cells were detected by real-time PCR.The results showed that Lnc₀02783 and miR-188-5p were related to the growth and development of goat skeletal muscle,and the expression trends of their had a certain negative relationship.（4）After interference with the expression of Lnc₀02783,qPCR detected the expression change of miR-188-5p.At the same time,the expression of Lnc₀02783 was detected after overexpression of miR-188-5p.It was found that the expression of miR-188-5p was down-regulated after interference with Lnc₀02783 expression,while the expression of Lnc₀02783 was down-regulated after overexpression of miR-188-5p.（5）After transfecting si-Lnc 002783 and si-NC for 24 h,the cells were induced to differentiate for 72 h,and the expression of Myog was detected by qPCR.The results showed that Lnc₀02783 can inhibit the differentiation of goat skeletal muscle satellite cells.In summary,the goat skeletal muscle satellite cells were successfully obtained in this experiment,and the selected Lnc₀02783 and miR-188-5p promoted and inhibited the proliferation of satellite cells,respectively.Lnc₀02783 and miR-188-5p are associated with goat muscle growth and development and there may be a targeted relationship between them.In addition,Lnc₀02783 has an inhibitory effect on the differentiation of goat skeletal muscle satellite cells.