外泌体介导细胞间通讯调控猪肌纤维代谢模式的DIA蛋白质组解析

针对育种中“商业猪种肉质欠佳，优良肉质性状形成遗传机制不清”的瓶颈问题，围绕我国猪品种改良的重大需求，以瘦肉型大白猪和脂肪型梅山猪的骨骼肌和脂肪组织来源的外泌体为研究对象，将探索猪骨骼肌和脂肪组织来源的外泌体对成肌细胞的影响；基于高精度、高灵敏度的非数据依赖式（DIA）蛋白质组学技术，首次尝试捕获猪骨骼肌和脂肪来源的外泌体蛋白质，并筛选出特异性调控肌纤维的蛋白质；从细胞间通讯调控的角度切入，对候选蛋白的功能和调控机制进一步确证。预期结果将获得精准解析的猪骨骼肌和脂肪来源的外泌体蛋白质数据，结合骨骼肌和脂肪来源的外泌体对骨骼肌的调控规律，建立起肌纤维生长和代谢等生物学事件中组织内与组织间外泌体的调控作用模型，阐明骨骼肌/脂肪来源的外泌体蛋白质介导细胞间通讯调控骨骼肌类型和代谢模式的分子机制，揭示细胞微环境塑造优质肉表型的遗传机理，为猪优良肉质性状的分子设计育种提供新的科学依据。

In view of the bottleneck problem "commercial pigs with poor meat quality and the genetic mechanisms of advanced meat quality remain unclear " in the pig breeding, and to boost of pig breeding industry in China, this research project aims to study the regulating functions and mechanisms of skeletal muscle and adipose tissue derived exosomes of Yorkshire pigs and Meishan pigs, the two representative breeds for lean and fatty meat production pigs. The whole proposal contains three aims: 1. Investigate the effects of skeletal muscle and adipose tissue derived exosomes on the myoblasts; 2. Profile the proteome of skeletal muscle-derived exosomes and adipose tissue-derived exosomes using Data-Independent Acquisition-Based quantitative proteomic analysis, and further identify exosome proteins with differential abundance between different developmental stages and breeds. 3. Clarify the functions and regulation mechanism of candidate exosome proteins which could regulate muscle fibers formation and metabolism through intercellular communication. By using Integrative proteomic analysis and functional validation, we will elucidate how exosome-mediated intercellular communication could affect pig muscle metabolism within and across tissues. The study will provide an alternative view for understanding the genetic mechanism of the meat quality traits that shaped by the cell microenvironment, and reveal the function and regulation mechanisms of exosomes in economically important biological events such as muscle fiber types and metabolism, which are the basis of molecular breeding for desired meat quality traits.