Peng Lingwei, Zhang Xiaolian, Du Yuqin, Li Fan, Han Jiazheng, Liu Oujin, Dai Shoulu, Zhang Xiang, Liu George E, Yang Liguo, Zhou Yang
Key Laboratory of Agricultural Animal Genetics, Breeding and Reproduction of Ministry of Education, Huazhong Agricultural University, Wuhan 430070, China.
Animal Genomics and Improvement Laboratory, BARC, USDA-ARS, Beltsville, MD 20705, USA.
iScience. 2023 Aug 28;26(10):107753. doi: 10.1016/j.isci.2023.107753. eCollection 2023 Oct 20.
We performed direct RNA sequencing (DRS) together with PCR-amplified cDNA long and short read sequencing for cattle adipocyte at different stages. We proved that the DRS was with advantages to avoid artificial transcripts and questionable exitrons. Totally, we obtained 68,124 transcripts with information of alternative splicing, poly (A) length and mRNA modification. The number of transcripts for adipogenesis was expanded by alternative splicing, which lead regulation mechanisms far more complex than ever known. We detected 891 differentially expressed genes (DEGs). However, 62.78% transcripts of DEGs were not significantly differentially expressed, and 248 transcripts showed opposite changing directions with their genes. The poly (A) tail became globally shorter in differentiated adipocyte than in primary adipocyte, and had a weak negative correlation with gene/transcript expression. Moreover, the study of different mRNA modifications implied their potential roles in gene expression and alternative splicing. Overall, our study promoted better understanding of adipogenesis mechanisms in cattle adipocytes.
我们对不同阶段的牛脂肪细胞进行了直接RNA测序（DRS）以及PCR扩增的cDNA长读长和短读长测序。我们证明了DRS具有避免人工转录本和可疑外显子的优势。总共，我们获得了68124个具有可变剪接、聚腺苷酸（poly (A)）长度和mRNA修饰信息的转录本。脂肪生成的转录本数量通过可变剪接得以扩展，这使得调控机制比以往所知的更加复杂。我们检测到891个差异表达基因（DEG）。然而，62.78%的DEG转录本没有显著差异表达，并且有248个转录本与其基因呈现相反的变化方向。与原代脂肪细胞相比，分化脂肪细胞中的聚腺苷酸（poly (A)）尾巴整体变短，并且与基因/转录本表达呈弱负相关。此外，对不同mRNA修饰的研究暗示了它们在基因表达和可变剪接中的潜在作用。总体而言，我们的研究促进了对牛脂肪细胞脂肪生成机制的更好理解。
