Carbohydrate-based interactions of oviductal sperm reservoir formation-studies in the pig.
Competitive inhibition of sperm to explants of the oviductal epithelium was used to study the complementary receptor system that may be involved in the establishment of the oviductal sperm reservoir in the pig. Sperm binding to the oviductal explants is expressed as Binding Index (BI = sperm cells/0.01 mm(2)). From a set of glycoproteins with known oligosaccharide structures, only asialofetuin and ovalbumin showed inhibitory activity, indicating that ovalbumin may block high affinity binding sites (IC(50) congruent with 1.3 microM) and asialofetuin low affinity sites (IC(50) congruent with 18 microM) of the complementary receptor systems, whereas fetuin carrying terminal sialic acid has no effect. Ovalbumin glycopeptides were isolated by Con A affinity chromatography and reverse-phase HPLC following tryptic digestion. Glycopeptides and enzymatically released glycans were analyzed by MS, and were shown to represent preferentially the two high mannose type glycans (Man)(5)(GlcNAc)(2) and (Man)(6)(GlcNAc)(2), and as a minor component the hybrid type glycan (Hex)(4)(GlcNAc)(5). Glycopeptides (84% inhibition) and glycans (81% inhibition) significantly reduced sperm-oviduct binding at a concentration of 3 microM, whereas the deglycosylated peptides showed no inhibitory activity. Mannopentaose (IC(50) congruent with 0.8 microM) representing the oligomannose residue of the high mannose glycans of ovalbumin was as effective as ovalbumin. These data indicate that the carbohydrate-based mechanisms underlying the formation of the oviductal sperm reservoir in the pig is the result of the concerted action of at least the high-affinity binding sites for oligomannose or nonreducing terminal mannose residues and low-affinity binding of galactose.
Wagner
Andrea
A
Institute of Reproductive Medicine, School of Veterinary Medicine Hannover, Germany.
Ekhlasi-Hundrieser
Mahnaz
M
Hettel
Christiane
C
Petrunkina
Anna
A
Waberski
Dagmar
D
Nimtz
Manfred
M
Töpfer-Petersen
Edda
E
D000818:Animals; D001212:Asialoglycoproteins / Q000378:metabolism*; D002240:Carbohydrate Sequence; D005187:Fallopian Tubes / Q000378:metabolism*; D005260:Female; D060748:Fetuins; D066298:In Vitro Techniques; D008297:Male; D008969:Molecular Sequence Data; D009844:Oligosaccharides / Q000737:chemistry / Q000378:metabolism; D010047:Ovalbumin / Q000378:metabolism*; D010446:Peptide Fragments / Q000378:metabolism; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization; D013094:Spermatozoa / Q000378:metabolism*; D013552:Swine / Q000502:physiology*; D000509:alpha-Fetoproteins / Q000378:metabolism*
D001212:Asialoglycoproteins; D060748:Fetuins; D009844:Oligosaccharides; D010446:Peptide Fragments; D000509:alpha-Fetoproteins; C017029:asialofetuin; D010047:Ovalbumin