GLYCOSAMINOGLYCANS ISOLATED FROM CAUDA EPIDIDYMAL FLUID IMPROVES SPERM FEATURES IN LIQUID-STORED PORCINE SPERMATOZOA

BARONIAN, D; OJEDA, G; BUTTAZZONI, AC; CANTU, DF; DE BLAS, G; SOSA, MA; AGUILERA ,AC

Mendoza

Congreso; XL Reunión anual de la sociedad de biología de cuyo; 2022

Sociedad de biologia de cuyo

Artificial insemination (AI) is the most practiced technique in porcine reproduction. Since this practice requires the storage of spermatozoa in semen extenders, there is an increasing interest in improving the liquid-storage composition to preserve sperm quality. Glycosaminoglycans (GAGs) are a family of linear polysaccharides comprised of repeating hexosamine-containing disaccharides that are found in the reproductive fluids along the male and female genital tracts. Different studies showed that GAGs may be involved in sperm processes such as capacitation and acrosome reaction in vivo. The aim of this study was to investigate the effect of GAGs isolated from cauda epididymal fluid (G-EF) and seminal plasma (G-SP) on viability, membrane stability, and mitochondrial activity of boar spermatozoa stored in commercial sperm extenders. GAGs from EF and SP were isolated by protease digestion, lipid extraction, and by different precipitation conditions. Fresh boar sperm was diluted in commercial short-term extender (BTS) and long-term extender (GM) supplemented with G-EF and G-SP, for 3 and 7 days at 16oC, respectively. Spermatozoa storage either with G-EF or G-SP showed a high percentage of membrane stability and viability assessed by flow cytometry using M540/YoPro (P < 0.05). Mitochondrial activity, assessed by Rhodamine123, showed a significant decrease in G-EF suggesting that GAGs could preserve mitochondrial function in liquid-stored spermatozoa (P < 0.05). To test the functional ability to undergo sperm capacitation, spermatozoa were incubated in TALP capacitation medium for 1 h at 38.5oC in a humid atmosphere. No differences were observed in the capacitation status assessed by flow cytometry using M540/YoPro (P < 0.05). These results provide evidence that the supplementation of GAGs from EF and SP might have a protective effect on boar spermatozoa increasing membrane stability, reducing mitochondrial activity, and maintaining viability during liquid-stored sperm storage.