Surface glycocalyx regulates cell-to-cell interactions and in male gametes is involved in sperm-oviductal epithelial cells adhesion during capacitation as well as in thè sperm-egg interaction. The aim of this study was to identify and localise the oligosaccharide sequences of the glycocalyx in stallion-ejaculated sperrnatozoa using tectin histochemistry. Semen from two trotters (three ejaculates each) was diluted in Tyrode's medium to remove seminal plasma, fixed in 4% (w/v) neutral formalin. smeared on slides coated with poly-L-lisine and incubated with appropriate dilmions of 11 lectins (n = 10 HRP-conjugates and n = 2 biotinylated). The lectins Con A and BSA I-B4 did not show a binding site, whereas the lectins MAL II,SNA,RCA120,HP A, and WGA gave a uniform labellmg of glycocalyx. In addition, the acrosomial region reacted with PNA and BSA II, whereas the post-acrosomal region and flagellimi bound HPA and UEA I. These findings indicate that: (1) glycocalyx contains oligosaccharides ending with Neu5Aca2,3Gal, Neu5Aca2,6Gal/Gal- NAc, lactosamine, and aGalNAc, (2) the acrosomal region of glycocalyx is characterized by oligosacchandes terminating with GalB1,3GalNAc or GlcNAc, (3) the post-acrosomal region and thè flagellum also display terminal a-L-fucose residues. Thiscarbohydrate pattern of the glycocalyx could play a role in the behaviour of spermatozoa during flow into the oviduct as well as in the interaction with oocytes during fertilization
Glycoconjugate expression on the plasma membrane of horse-ejaculated spermatozoa revealed by lectin histochemistry
DESANTIS, Salvatore;VALENTINI, Luisa;VENTRIGLIA G;
2004
Abstract
Surface glycocalyx regulates cell-to-cell interactions and in male gametes is involved in sperm-oviductal epithelial cells adhesion during capacitation as well as in thè sperm-egg interaction. The aim of this study was to identify and localise the oligosaccharide sequences of the glycocalyx in stallion-ejaculated sperrnatozoa using tectin histochemistry. Semen from two trotters (three ejaculates each) was diluted in Tyrode's medium to remove seminal plasma, fixed in 4% (w/v) neutral formalin. smeared on slides coated with poly-L-lisine and incubated with appropriate dilmions of 11 lectins (n = 10 HRP-conjugates and n = 2 biotinylated). The lectins Con A and BSA I-B4 did not show a binding site, whereas the lectins MAL II,SNA,RCA120,HP A, and WGA gave a uniform labellmg of glycocalyx. In addition, the acrosomial region reacted with PNA and BSA II, whereas the post-acrosomal region and flagellimi bound HPA and UEA I. These findings indicate that: (1) glycocalyx contains oligosaccharides ending with Neu5Aca2,3Gal, Neu5Aca2,6Gal/Gal- NAc, lactosamine, and aGalNAc, (2) the acrosomal region of glycocalyx is characterized by oligosacchandes terminating with GalB1,3GalNAc or GlcNAc, (3) the post-acrosomal region and thè flagellum also display terminal a-L-fucose residues. Thiscarbohydrate pattern of the glycocalyx could play a role in the behaviour of spermatozoa during flow into the oviduct as well as in the interaction with oocytes during fertilizationI documenti in IRIS sono protetti da copyright e tutti i diritti sono riservati, salvo diversa indicazione.
