基于单细胞转录组学的滩羊肌内脂肪细胞-肌卫星细胞互作机制研究

Fresh meat, light mutton flavor, and high protein content are the merits of Tan Sheep. However, due to the degeneration of its own species, the development of Tan sheep industry is affected to a large extent by its low productivity of meat and low intramuscular fat. The process of muscle development and fat deposition in Tan sheep involves the expression of large numbers of genes and their regulation of network. At present, that the molecular regulation mechanism of muscle development and fat deposition in Tan sheep is still not clear, thus it is necessary to further explore the related genes and study their function and regulation mechanism. This project will focus on Ningxia Tan sheep as the research object, establishco-culture system of preadipocytes and myogenic satellite cells, and sequence differentiating cells co-cultured or cultured solely in vitro by whole transcriptome (RNA-seq and small RNA-seq) high-throughput sequencing (HTS) technologies. These technologiescould be used to identify the differentially expressed mRNAs, lncRNAs, circRNAs, miRNAs during the proliferation and the differentiation stages. In addition, these technologies could also be used to predict target genes, enrich to the GO terms and KEGG pathways, which can analyze the functions of the differentially expressed genes. This study not only provides a new idea for understanding the developmental regularities of adipocytes and myocytes, but also supplies a foundation to further study transcriptomics and optimize the quality of Tan sheep mutton.

滩羊肉质鲜嫩，膻味轻，但其产肉率低、肌内脂肪分布较少，在很大程度上影响滩羊产业发展。绵羊肌肉发育与脂肪沉积过程涉及大批基因表达及网络式调控。因此，有必要进一步的发掘相关基因，对其功能和调控机理进行研究。本项目拟以宁夏滩羊为研究对象，利用细胞体外培养技术建立滩羊肌内前体脂肪细胞-肌卫星细胞联合培养体系，并利用全转录组高通量测序（RNA-seq and small RNA-seq）技术对不同分化阶段联合培养细胞及对照组单独培养细胞进行全转录组高通量测序，筛选其增殖和分化过程中差异表达的mRNAs、lncRNAs、circRNAs和miRNAs，并预测主要信号转导通路及其复杂调控网络与基因共表达网络，分析肌内脂肪细胞与肌细胞相互影响与作用机制。本研究不仅为探讨脂肪细胞与肌细胞发育规律及相互作用提供新的思路，也为进一步选育优质肉用滩羊品种，改善滩羊肉品质奠定基础。

滩羊肉质鲜嫩，膻味轻，但其产肉率低、肌内脂肪分布较少，在很大程度上影响滩羊产业发展。绵羊肌肉发育与脂肪沉积过程涉及大批基因表达及网络式调控。因此，有必要进一步的发掘相关基因，对其功能和调控机理进行研究。本项目拟以宁夏滩羊为研究对象，利用细胞体外培养技术建立滩羊肌内前体脂肪细胞-肌卫星细胞联合培养体系，并利用全转录组高通量测序（RNA-seq and small RNA-seq）技术对不同分化阶段联合培养细胞及对照组单独培养细胞进行全转录组高通量测序，筛选其增殖和分化过程中差异表达的mRNAs、lncRNAs、circRNAs和miRNAs，并预测主要信号转导通路及其复杂调控网络与基因共表达网络，分析肌内脂肪细胞与肌细胞相互影响与作用机制。其中，项目完成的主要研究内容包括：（1）建立了滩羊肌内前体脂肪细胞-骨骼肌卫星细胞联合培养体系；（2）结合RNA-Seq，研究了滩羊肌内前体脂肪细胞增殖和分化的作用及分子机制；（3）结合RNA-Seq，研究了滩羊肌卫星细胞增殖和分化的作用及分子机制；（4）结合RNA-Seq，滩羊肌内前体脂肪细胞与骨骼肌卫星细胞互作分子机制。以上研究不仅为探讨脂肪细胞与肌细胞发育规律及相互作用提供新的思路，也为进一步选育优质肉用滩羊品种，改善滩羊肉品质奠定基础。