Chicken seminal fluid lacks CD9- and CD44-bearing extracellular vesicles |
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| Authors: | Manuel Alvarez-Rodriguez Maria Ntzouni Dominic Wright Kabirul Islam Khan Manel López-Béjar Cristina A Martinez Heriberto Rodriguez-Martinez |
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| Institution: | 1. Department of Biomedical and Clinical Services (BKV), BHK/O&G Linköping University, Linköping, Sweden;2. Microscopy Unit, Faculty of Medicine and Health Sciences (MEDFAK), Core Facility (COREF) Linköping University, Linköping, Sweden;3. Department of Physics, Chemistry and Biology, Faculty of Science and Engineering, Linköping University, Linköping, Sweden;4. Department of Genetics and Animal Breeding, Chattogram Veterinary and Animal Sciences University, Chattogram, Bangladesh;5. Department of Animal Health and Anatomy, Faculty of Veterinary, Universitat Autòmoma de Barcelona, Cerdanyola del Vallès, Barcelona, Spain |
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| Abstract: | The avian seminal fluid (SF) is a protein-rich fluid, derived from the testis, the rudimentary epididymis and, finally, from the cloacal gland. The SF interacts with spermatozoa and the inner cell lining of the female genital tract, to modulate sperm functions and female immune responsiveness. Its complex proteome might either be free or linked to extracellular vesicles (EVs) as it is the case in mammals, where EVs depict the tetraspanin CD9; and where those EVs derived from the epididymis (epididymosomes) also present the receptor CD44. In the present study, sperm-free SF from Red Jungle Fowl, White Leghorn and an advanced intercross (AIL, 12th generation) were studied using flow cytometry of the membrane marker tetraspanin CD9, Western blotting of the membrane receptor CD44 and electron microscopy in non-enriched (whole SF) or enriched fractions obtained by precipitation using a commercial kit (Total Exosome Precipitation Solution). Neither CD9- nor CD44 could be detected, and the ultrastructure confirmed the relative absence of EVs, raising the possibility that avian SF interacts differently with the female genitalia as compared to the seminal plasma of mammals. |
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| Keywords: | CD44 CD9 electron microscopy exosome precipitation rooster seminal fluid tetraspanins western blot |
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