Extracellular vesicles isolated from porcine seminal plasma exhibit different tetraspanin expression profiles

• Published in: Scientific reports 2019-08, Vol.9 (1), p.11584-9, Article 11584
• Main Authors: Barranco, Isabel, Padilla, Lorena, Parrilla, Inmaculada, Álvarez-Barrientos, Alberto, Pérez-Patiño, Cristina, Peña, Fernando J., Martínez, Emilio A., Rodriguez-Martínez, Heriberto, Roca, Jordi
• Format: Article
• Language: English
• Subjects: 101/1; 13; 13/31; 14; 14/28; 631/1647/328/2082; 631/443/494; Animals; CD63 antigen; CD81 antigen; CD9 antigen; Electron microscopy; Exosomes; Exosomes - chemistry; Exosomes - ultrastructure; Extracellular vesicles; Extracellular Vesicles - chemistry; Extracellular Vesicles - ultrastructure; Flow Cytometry; Humanities and Social Sciences; Light scattering; Male; Morphology; multidisciplinary; Science; Science (multidisciplinary); Semen; Semen - chemistry; Seminal vesicle; Swine; Swine - metabolism; Tetraspanin 28 - analysis; Tetraspanin 29 - analysis; Tetraspanin 30 - analysis; Tetraspanins - analysis; Ultracentrifugation
• ISSN: 2045-2322; 2045-2322
• DOI: 10.1038/s41598-019-48095-3

Seminal extracellular vesicles (EVs) include exosomes (ø 40–120 nm) and microvesicles (MVs, ø 120–1000 nm), which would be involved in multiple functional reproductive roles. The study aimed to establish which EV subtypes are present in pig semen, using a high-resolution flow cytometer to explore differences in their tetraspanin expression profile. The EVs were isolated from 12 pig ejaculates using serial ultracentrifugation and characterized by dynamic light scattering and electron microscopy for size and morphology as well as for tetraspanin expression using flow cytometry with Carboxyfluorescein succinimidyl ester (CFSE) and antibodies against CD9, CD63 and CD81. Pig semen contained a heterogeneous EV-population regarding size and morphology. Flow cytometric analysis demonstrated that the proportion of EVs expressing CD63 and CD9 was higher in MVs ( P