Anti-hyaluronidase oligosaccharide derived from chondroitin sulfate a effectively reduces polyspermy during in vitro fertilization of porcine oocytes.
The present study was conducted to examine the effects of chondroitin sulfate A-derived oligosaccharide (ChSAO) on hyaluronidase activity and in vitro fertilization (IVF) parameters. The activity of hyaluronidase extracted from preincubated boar sperm was completely blocked by ChSAO at concentrations of 10 microg/ml or higher. After in vitro maturation of porcine cumulus-oocyte complexes, some oocytes were freed from their cumulus cells, and cumulus-intact or cumulus-free oocytes were inseminated with sperm in IVF medium containing various concentrations of ChSAO (0.1-100 microg/ml). In cumulus-intact oocytes, the penetration and the polyspermy rates (39% and 28%, respectively) were significantly decreased by treatment with 100 microg/ml ChSAO compared with those of oocytes treated without ChSAO (63% and 52%, respectively). On the contrary, in cumulus-free oocytes, the addition of 10-100 microg/ml ChSAO significantly reduced the polyspermy rate compared with the control (25-30% versus 53%, respectively), whereas ChSAO had no effect on sperm penetration. Interestingly, ChSAO added to IVF medium significantly decreased the number of sperm bound to the zona pellucida (ZP) of cumulus-free oocytes in a concentration-dependent manner between 0.1 and 100 microg/ml. However, ChSAO had no effect on the time course change in ZP modification after oocyte activation by electrostimulation and the incidence of the acrosome-reacted sperm. Treatment with 100 microg/ml ChSAO during IVF of cumulus-free oocytes significantly increased the proportion of development to the blastocyst stage after in vitro insemination. Therefore, the present findings indicate that hyaluronidase-inhibitory ChSAO is an efficient probe for promoting normal fertilization process in terms of an effective decrease in the incidence of polyspermy during IVF of porcine oocytes.
Tatemoto
Hideki
H
Department of Bioproduction, Faculty of Agriculture, University of the Ryukyus, Nishihara-cho, Okinawa 903-0213, Japan. hidettmt@agr.uryukyu.ac.jp
Muto
Norio
N
Yim
Sun-Deok
SD
Nakada
Tadashi
T
D020101:Acrosome Reaction / Q000187:drug effects; D000818:Animals; D001755:Blastocyst / Q000502:physiology; D002478:Cells, Cultured; D002809:Chondroitin Sulfates / Q000737:chemistry*; D004527:Egg Proteins / Q000187:drug effects / Q000378:metabolism; D047108:Embryonic Development / Q000187:drug effects; D004791:Enzyme Inhibitors / Q000494:pharmacology; D005260:Female; D005307:Fertilization in Vitro / Q000379:methods; D006821:Hyaluronoglucosaminidase / Q000037:antagonists & inhibitors* / Q000378:metabolism; D008297:Male; D008562:Membrane Glycoproteins / Q000187:drug effects / Q000378:metabolism; D009844:Oligosaccharides; D009865:Oocytes / Q000187:drug effects* / Q000502:physiology; D011956:Receptors, Cell Surface / Q000187:drug effects / Q000378:metabolism; D013075:Sperm Capacitation / Q000187:drug effects; D013084:Sperm-Ovum Interactions / Q000187:drug effects*; D013552:Swine; D015044:Zona Pellucida / Q000187:drug effects / Q000378:metabolism; D000072216:Zona Pellucida Glycoproteins
D004527:Egg Proteins; D004791:Enzyme Inhibitors; D008562:Membrane Glycoproteins; D009844:Oligosaccharides; D011956:Receptors, Cell Surface; D000072216:Zona Pellucida Glycoproteins; D002809:Chondroitin Sulfates; D006821:Hyaluronoglucosaminidase