Effect of Monosaccharide L-fucose and Polysaccharide Fucoidan on Sperm ??L-fucosidase Activity and Relation to Sperm-oocyte Interaction in Pig

Abstract

Carbohydrate-protein interactions are known to be important in gamete interactions. Several evidence indicated that a fucose-containing sulfated polysaccharide fucoidan was potential inhibitor of fertilization in vitro and thus fucose seemed to be part of the recognition signal of gamete interaction in mammals. In recent investigation we found that α-L-fucosidase activity was present in boar spermatozoa and it was related to sperm binding to and penetration into zona pellucida (ZP) in vitro. The objective of this study was to determine the effects of monosaccharide L-fucose and polysaccharide fucoidan on sperm α-L-fucosidase activity and relation to sperm-oocyte interaction in pig. Results indicated that the activity of sperm α-L-fucosidase was largely inhibited (62%) when sperm suspension was treated with monosaccharide L-fucose. It also significantly inhibited the number of sperm binding to ZP (32%) and penetration into zona-intact oocytes (72%), but did not inhibit penetration into zona-free oocytes when fertilization medium contained Lfucose. The chlorotetracycline (CTC) assessment showed that L-fucose did not affect induction of sperm capacitation and acrosome reaction. In contrast, the activity of sperm α-L-fucosidase was not inhibited when sperm suspension was treated with polysaccharide fucoidan but sperm-ZP binding was greatly inhibited (85%) and completely blocked sperm penetration into zona-intact or zona-free oocytes. The CTC assessment showed that fucoidan increased the F pattern and decreased the AR pattern sperm. These results suggested that the different inhibitory mechanisms were present between monosaccharide L-fucose and polysaccharide fucoidan on sperm-oocyte interaction, the inhibition effect of α-L-fucose on sperm binding and penetrating into ZP caused sperm α-L-fucosidase inhibited by α-Lfucose.

Assessment of buffalo semen with the 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide reduction assay

“…Yellow 3-(4,5-dimethylthiazol-2-y1)-2,5-diphenyltetrazolium bromide (MTT), a water-soluble tetrazolium salt dye, is converted to water-insoluble purple formazan by the succinate dehydrogenase system of an active mitochondria by the reductive cleavage of its tetrazolium ring (Slater et al, 1963). Thus, the amount of formazan formed can be determined spectrophotometrically and can serve as an estimate of the number of active mitochondria, and hence the viable cells, in a sample (Denizot and Lang, 1986;Song et al, 2007).…”

α-L-fucosidase enhances capacitation-associated events in porcine spermatozoa

“…2008). Addition of fucose and fucoidan to the fertilisation media resulted in a significant decrease in penetration rates in pigs and cows (Tanghe et al, 2004;Song et al, 2007b). It was surprising that fucosidase reduced the SPZ-ZP binding only after a 120 min incubation period.…”

“…Its substrate, fucose, has been detected in sugar chains on the spermatozoal surface in the mouse (Boldt et al, 1989), bull (Taitzoglou et al, 2007) and boar (Jiménez et al, 2003), and also on the inner region of the ZP in pigs (Song et al, 1999). In addition, when carbohydrate involvement in spermatozoaegg interactions were studied by adding sugars to fertilisation media, fucose and the polysaccharide fucoidan induced a significant drop in penetration rates in cattle (Tanghe et al, 2004) and pigs (Töpfer-Petersen et al, 1985;Song et al, 2007b). Likewise, glycan-binding enzymes in the spermatozoal membrane play a role in spermatozoa-oocyte binding through interaction with complementary glycan units on the surface of the ZP (Tulsiani, 2000).…”

“…α-L-fucosidase is present in bull (Zahler and Doak, 1975;Jauhiainen and Vanha-Perttula, 1986), boar (Song et al, 2000), human (Venditti et al, 2007;Venditti and Bean, 2009), hamster (Venditti et al, 2010) and mouse spermatozoa (Phopin et al, 2012(Phopin et al, , 2013. Since α-L-fucosyl residues in the ZP and α-L-fucosidase in spermatozoa are involved in in vitro spermatozoa-oocyte interactions in pigs, a hydrolytic role of the spermatozoal enzyme during ZP penetration has been proposed (Song et al, 2007b).…”

Evaluation of Boar Sperm Viability by MTT Reduction Assay in Beltsville Thawing Solution Extender

“…Succinate dehydrogenase system of active mitochondria converts this dye to water-insoluble purple formazan on the reductive cleavage of its tetrazolium ring (Slater et al, 1963). Thus, the amount of formazan formed can be determined spectrophotometrically and serves as an estimate of the number of mitochondria and hence the number of living cells in the sample (Denizot and Lang, 1986;Song et al, 2007). In many studies, MTT assay was used which were related to viability of different cells (Mosmann, 1983;Levitz and Diamone, 1985;Carmichael et al, 1987;Campling et al, 1988;Freimoser et al, 1999).…”