Characterization of the exosomes in the allantoic fluid of the chicken embryo
The embryo stage is critical for chicken development. Numerous studies have been conducted to clarify the dynamic changes and functions of various proteins and the composition of amino acids during embryo development. However, the physiological characteristics of extraembryonic fluid (allantoic and amniotic), especially allantoic fluid (AF), remain largely unexplored; furthermore, how information is transmitted from embryonic fluid is unknown. In this study, AF-derived exosomes ranging from 60 to 160 nm in diameter from fertilized eggs at 13 days of incubation of fast-growth chickens (WG chicken) medium-growth chickens (Silky N4 chicken), and slow-growth chickens (Huiyang Beard chicken), were isolated and purified by different ultra-centrifugations and further verified by transmission electron microscopy (TEM) and a flow nano-analyzer. Expression of the exosomal positive biomarkers of ALIX and HSP70 as well as lack of the epithelium marker GRP78, were observed by western blotting. In addition, small RNA sequencing revealed that AF-derived exosomes at 13 days of incubation contained a large number of known miRNAs (32.62–65.83%). The top 10 most abundant and co-expressed miRNAs were primarily related to development, growth and immunity. In addition, AF-derived exosomes promoted DF-1 cell migration. These findings broadened our understanding of the characteristic of AF-derived exosomes.