Identification of perivitelline N-linked glycans as mediators of sperm-egg interaction in chickens.
This study demonstrates that carbohydrates play an essential role in sperm-egg interactions in birds. Sperm-egg interaction was measured in vitro as the ability of spermatozoa to hydrolyse a small hole in the inner perivitelline layer, the equivalent of the mammalian zona pellucida. Preincubation with Triticum vulgaris lectin (WGA) and succinyl-WGA (S-WGA) at 10 microgram ml(-1) resulted in complete inhibition of sperm-egg interaction, whereas at the same concentration a range of other lectins (Canavalia ensiformis (Con A), Arachis hypogea (PNA), Ulex europaeus II (UEA II), Solanum tuberosum (STA), Tetragonolobus purpureas (LTA) and Pisum sativum (PSA)) were unable to inhibit sperm egg interaction significantly, although fluorescein-labelled derivatives of these lectins were found to stain the inner perivitelline layer. Significant inhibition of sperm-egg interaction was achieved by the addition of N-acetyl-D-glucosamine and fucoidin to the assay mixture; however, D-glucose, D-galactose, D-fucose and L-fucose had no significant effect on sperm-egg interaction. Pretreatment of the inner perivitelline layer with N-glycanase significantly reduced sperm-egg interaction, whereas treatment with O-glycanase had no effect. These results demonstrate that N-linked glycans play an essential role in sperm-egg interaction in chickens.
Robertson
L
L
Avian Reproduction Group, School of Science and Engineering, University of Abertay Dundee, Bell Street, Dundee DD1 1HG, UK.
Wishart
G J
GJ
Horrocks
A J
AJ
D000117:Acetylglucosamine / Q000494:pharmacology*; D000581:Amidohydrolases / Q000494:pharmacology*; D000818:Animals; D002478:Cells, Cultured; D002645:Chickens / Q000502:physiology*; D046508:Culture Techniques; D005260:Female; D006596:Hexosaminidases / Q000494:pharmacology; D006868:Hydrolysis; D037102:Lectins / Q000494:pharmacology; D008297:Male; D009005:Monosaccharides / Q000494:pharmacology; D043524:Peptide-N4-(N-acetyl-beta-glucosaminyl) Asparagine Amidase; D011134:Polysaccharides / Q000378:metabolism* / Q000494:pharmacology; D011485:Protein Binding; D013084:Sperm-Ovum Interactions / Q000187:drug effects*; D014817:Vitelline Membrane / Q000378:metabolism*; D014909:Wheat Germ Agglutinins / Q000494:pharmacology
D037102:Lectins; D009005:Monosaccharides; D011134:Polysaccharides; D014909:Wheat Germ Agglutinins; C007789:fucoidan; D006596:Hexosaminidases; C060638:glycopeptide alpha-N-acetylgalactosaminidase; D000581:Amidohydrolases; D043524:Peptide-N4-(N-acetyl-beta-glucosaminyl) Asparagine Amidase; D000117:Acetylglucosamine