Sperm-zona pellucida interaction involves a carbonyl reductase activity in the hamster.
For successful fertilization to occur, the spermatozoa must transit through an egg-specific extracellular matrix or zona pellucida (zp) to reach and ultimately fuse with the oocyte plasma membrane. This process involves ligand-receptor recognition between the zp and the acrosomal cap of the sperm. The hamster sperm protein P26h, a receptor which is acquired during epididymal transit, has been suggested to act in sperm-zp binding. The cloning and characterization of the full-length cDNA-encoding hamster P26h revealed 85% identity with a porcine lung carbonyl reductase. To better understand the mechanism by which P26h interacts with zp proteins, we investigated carbonyl reductase activity during gamete interactions. In the present study, we show that specific inhibitors of carbonyl reductase such as diclofenac and phenylbutazone decreases sperm-zp binding without affecting the motility, progressivity or acrosome integrity of sperm. We also detected, and partly purified, carbonyl reductase activities from cauda epididymal sperm protein extract and this activity was associated with an enriched fraction of P26h. Removing P26h from the partly purified protein fractions by immunoaffinity chromatography led to the loss of carbonyl reductase activity. The findings that sperm-zp binding is blocked by carbonyl reductase inhibitors and that P26h is active in mature sperm suggest that P26h could play an important role in the fertilization process.
Montfort
Lucile
L
Centre de Recherche en Biologie de la Reproduction, Département d'Obstétrique-Gynécologique, Faculté de Médecine, Université Laval, Ste-Foy, Quebec, Canada.
Frenette
Gilles
G
Sullivan
Robert
R
D000429:Alcohol Oxidoreductases / Q000037:antagonists & inhibitors / Q000378:metabolism*; D000818:Animals; D015153:Blotting, Western; D002448:Cell Adhesion; D015815:Cell Adhesion Molecules / Q000378:metabolism; D002846:Chromatography, Affinity; D006224:Cricetinae; D016861:Cyclooxygenase Inhibitors / Q000494:pharmacology; D004008:Diclofenac / Q000494:pharmacology; D005260:Female; D005306:Fertilization / Q000502:physiology*; D008297:Male; D010653:Phenylbutazone / Q000494:pharmacology; D013079:Sperm Maturation / Q000187:drug effects; D013081:Sperm Motility / Q000187:drug effects; D013094:Spermatozoa / Q000166:cytology / Q000201:enzymology* / Q000378:metabolism*; D015044:Zona Pellucida / Q000201:enzymology* / Q000378:metabolism*
D015815:Cell Adhesion Molecules; D016861:Cyclooxygenase Inhibitors; C118288:sperm antigen P26h; D004008:Diclofenac; D000429:Alcohol Oxidoreductases; D010653:Phenylbutazone