Key Laboratory of Agricultural Animal Genetics, Breeding and Reproduction, Ministry of Education, Wuhan, China.
College of Animal Science and Technology and College of Veterinary Medicine, Huazhong Agricultural University, Wuhan, China.
Reprod Domest Anim. 2022 Jan;57(1):98-110. doi: 10.1111/rda.14033. Epub 2021 Nov 12.
In mammals, seminal plasma extracellular vesicles (SPEVs) can regulate sperm motility and capacitation. The characteristics and functions of SPEVs in avians have been rarely reported. In this study, chicken SPEVs were isolated and characterized by transmission and scanning electron microscopy (TEM/SEM) and nanoparticle tracking analysis (NTA); furthermore, seven extracellular vesicle (EVs) marker proteins were detected by Western blot (WB). TEM revealed that chicken SPEVs had a classic bilayer membrane structure. NTA confirmed that the size of SPEVs was 30-250 nm, and concentration ranged from 8.0 E + 11-8.5 E + 11 particles/ml. There were 3073 SPEVs proteins identified by deep sequencing, including 2794 intracellular proteins and 279 extracellular proteins. The overlap rate of proteomes between chicken SPEVs and vesicles reported in the Vesiclepedia database reached 86%, and 360 new proteins that had not been reported by the ExoCarta and Vesiclepedia databases were identified in chicken SPEV proteomes. Gene Ontology (GO) analysis revealed that chicken SPEV proteins were mainly enriched in supplying energy and transporting protein. There were 4 IFT family proteins speculated to play an important role in sperm composition and function. Our data were compared with two previously published studies on the proteomics of chicken seminal plasma (SP) and hen uterine fluid, and some overlapping proteins described in chicken SPEVs had been identified in hen uterine fluid (545) and chicken SP (284). In conclusion, these findings will increase our understanding of the content and composition of proteome in SPEVs and provide new insights into the important role of the SPEV regulation in sperm functions.
在哺乳动物中，精液外泌体（SPEVs）可以调节精子的运动和获能。禽类 SPEVs 的特征和功能鲜有报道。本研究通过透射电子显微镜（TEM）/扫描电子显微镜（SEM）和纳米颗粒跟踪分析（NTA）分离和鉴定鸡 SPEVs，并通过 Western blot（WB）检测七种外泌体（EVs）标记蛋白。TEM 显示鸡 SPEVs 具有典型的双层膜结构。NTA 证实 SPEVs 的大小为 30-250nm，浓度范围为 8.0E+11-8.5E+11 个颗粒/ml。通过深度测序鉴定出 3073 种 SPEVs 蛋白，包括 2794 种细胞内蛋白和 279 种细胞外蛋白。鸡 SPEVs 与 Vesiclepedia 数据库中报道的其他禽类外泌体的蛋白质组重叠率达到 86%，在鸡 SPEV 蛋白质组中还鉴定出 360 种 ExoCarta 和 Vesiclepedia 数据库中未报道的新蛋白。GO 分析表明，鸡 SPEV 蛋白主要富集于提供能量和转运蛋白。有 4 种 IFT 家族蛋白推测在精子组成和功能中发挥重要作用。我们的数据与之前发表的关于鸡精液（SP）和母鸡子宫液蛋白质组学的两项研究进行了比较，在母鸡子宫液（545）和鸡 SP（284）中也鉴定到了一些在鸡 SPEVs 中描述的重叠蛋白。总之，这些发现将增加我们对 SPEVs 中蛋白质组内容和组成的理解，并为 SPEV 调节精子功能的重要作用提供新的见解。
