Abstract
Seminal plasma ingredients are important for maintenance of sperm viability. This study focuses on the effect of boar seminal plasma exosomes on sperm function during long-term liquid storage. Boar seminal plasma exosomes had typical nano-structure morphology as measured by scanning electron microscopy (SEM) and molecular markers such as AWN, CD9 and CD63 by western blot analysis. The effect on sperm parameters of adding different ratio of boar seminal plasma exosomes to boar sperm preparations was analyzed. Compared to the diluent without exosomes, the diluent with four times or sixteen times exosomes compared to original semen had higher sperm motility, prolonged effective survival time, improved sperm plasma membrane integrity (p < 0.05), increased total antioxidant capacity (T-AOC) activity and decreased malondialdehyde (MDA) content. The diluent containing four times concentration of exosomes compared to original semen was determined to inhibit premature capacitation, but not to influence capacitation induced in vitro. Inhibition of premature capacitation is likely related to the concentration of exosomes which had been demonstrated to transfer proteins including AWN and PSP-1 into sperm. In addition, using fluorescence microscopy and scanning electron microscopy analysis, it was demonstrated that exosomes in diluent were directly binding to the membrane of sperm head which could improve sperm plasma membrane integrity.
Highlights
In order to become fully fertile after leaving testis, mammalian sperm must undergo morphological and functional changes during transit through epididymis and other accessory sex organs
Boar seminal plasma exosomes prolonged effective motility time of sperm After the particle concentration was adjusted to the exosomes concentration of original seminal plasma, the effect of different concentrations of exosomes on boar sperm motility during liquid storage at 17°C was measured (Table 1)
These results indicated that Exo-4 and Exo-16 exosomes supplementation resulted in significantly prolonged sperm motility during storage
Summary
In order to become fully fertile after leaving testis, mammalian sperm must undergo morphological and functional changes during transit through epididymis and other accessory sex organs. Spermatozoa are mixed with another set of surface re-modeling components derived from the accessory sex glands. It is thought that early capacitation-related events are accompanied with the loss, modification and redistribution of molecules on the sperm surface during sperm storage in vitro [5, 6]. It has been observed that addition of ram or boar seminal plasma to sperm preparations improved viability and motility and reduced capacitation-like changes during sperm storage. It is necessary to identify the components of boar seminal plasma that help to maintain sperm function during sperm storage in vitro
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