Screening And Identification Of Long Noncoding Rnas In Adipose Tissue Of Qinchuan Cattle And Exploring Their Expression Specificity

All along,beef has been welcomed by consumers because of its high protein and low cholesterol.Beef tenderness affected by intramuscular fat or marble pattern not only affects the palatability of beef,but also is a key factor in determining consumer choice.The promotion of intermuscular fat production can increase the marble pattern,reduce the number of muscle fibers and thus increase beef tenderness.Beef tenderness is regulated by various factors including variety,nutrition,and environment.Therefore,genetic improvement of breeds can significantly improve beef meat quality.Bovine adipogenesis is a complex process that is affected by various regulatory factors,action pathways,and regulatory networks.The present study has a clearer understanding of this biological pathway.Numerous studies have shown that PPAR? and C/EBP? are the core which play a key role in this process.With the deepening of research,many recent studies have shown that lncRNAs also play a key role in the regulation of adipocyte development.However,they are mainly concentrated in humans and mice,and due to the poor conservation of lncRNA species,the founds of functional lncRNAs on mice cannot be directly applied to other species,and there are little studies on lncRNAs in cattle.Therefore,in this study,high-throughput sequencing of ribosomal RNA was used to analyze and screen the lncRNAs which only express in adipose tissue from tissues of muscle,fat,heart,liver,spleen,kidney,lung,and abomasum of Qinchuan cattle and verification of the results in fat,muscle,heart,liver,spleen,kidney,lung,large intestine,small intestine,rumen,reticulum,valve-stomach,abomasum,and testis of Qinchuan cattle calves by using real-time fluorescence quantitative PCR techniques.Among the identified lnc RNAs,we selected the lncRNA39520 with the highest expression and most specific adipose tissue for further study,namely the exploration of lncRNA39520 in Qinchuan cattle calves and 6 months,12 months,and 18 months of age.,24-month-old,60-month-old Qinchuan cattle adipose tissue expression,and 0d,3d,6d,9d,12 d expression in induced differentiation of Qinchuan cattle precursor adipocytes.The main results were as follows:1.Eight samples of the longissimus dorsi muscle,subcutaneous fat,heart,liver,spleen,kidney,lung,and abomasum of Qinchuan cattle were sampled.The transcriptome was sequenced and the sequencing data was analyzed for quality.The results showed that about 90% of reads can be compared to the reference genome,and 99% of the data is high quality data.Therefore,we can be sure that the sequencing data obtained is true and reliable.Can be used for the next study.2.Explore the location of these reads,their distribution on chromosomes,and their expression levels and expression differences.3.Transcripts were further screened.Long-chain non-coding RNAs expressed in various tissues were obtained.The differential expression and expression levels of long-chain non-coding RNAs were explored and 17 long-chain non-coding RNAs specifically expressed in adipose tissue were found.4.The long-chain non-coding RNA 39520 gradually increased in adipose tissue of calves,6 months,and 12 months of age in Qinchuan beef cattle,and reached the highest level in adipose tissue of Qinchuan beef cattle at 12 months of age.The lowest amount of adipose tissue was found in beef calves.5.The expression of long-chain non-coding RNA39520 gradually increased on the 0th to the 3rd day of differentiation of Qinchuan beef bovine preadipocytes,reached the highest on the third day,and had significant differences from the 0th day and the 12 th day.Since then,the amount of expression gradually decreased.In summary,this study successfully sampled the tissues of longissimus dorsi muscle,subcutaneous fat,heart,liver,spleen,kidney,lung,and abomasum of Qinchuan cattle calves and performed transcriptome sequencing.Long-chain non-coding RNAs specifically expressed in tissues were identified,and the long-chain non-coding RNA 39520 was selected for further investigation.It was explored in different stages of Qinchuan cattle adipose tissue and induced differentiation of Qinchuan cattle precursor fat cells.The characteristics of expression laid the foundation for further research on its functions.