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Regular article |
a Department of Animal Science,
b Department of Obstetrics and Gynecology, and
c Department of Biochemistry and Molecular Biology, University of Florida, Gainesville, Florida 32610–0294
d Department of Animal Science, University of Missouri-Columbia, Columbia, Missouri 65211
ABSTRACT
This study evaluated the effects of porcine oviduct-specific glycoprotein (pOSP) on in vitro fertilization (IVF), polyspermy, and development to blastocyst. Experiment 1 evaluated the effects of various concentrations (0–100 µg/ml) of purified pOSP on fertilization parameters, including penetration, polyspermy, male pronuclear formation, and mean number of sperm penetrated per oocyte. Experiment 2 examined the ability of an anti-pOSP immunoglobulin G to inhibit the observed effects of pOSP on fertilization parameters. Experiments 3 and 4 examined various concentrations of pOSP (0–100 µg/ml) on zona pellucida solubility and sperm binding, respectively. Lastly, experiment 5 assessed the effects of various concentrations of pOSP (0–100 µg/ml) on the in vitro embryo cleavage rate and development to blastocyst. Pig oocytes matured and fertilized in vitro were used for all experiments. An effect of treatment (P < 0.05) was detected for pOSP on penetration, polyspermy, and mean number of sperm per oocyte. Concentrations for pOSP of 0–50 µg/ml had no effect on sperm penetration rates; however, compared with the control, 100 µg/ml significantly decreased the penetration rate (74% vs. 41%). Addition of 10–100 µg/ml significantly reduced the polyspermy rate compared with the control (61% vs. 24–29%). The decrease in polyspermy achieved by addition of pOSP during preincubation and IVF was blocked with a specific antibody to pOSP. No effect of treatment was observed on zona digestion time relative to the control; however, the number of sperm bound to the zona pellucida was significantly decreased by treatment (P < 0.05). Compared with the control, all concentrations of pOSP examined reduced the number of sperm bound per oocyte (45 vs. 19–34). A treatment effect (P < 0.05) was observed for pOSP on embryo development to blastocyst but not on cleavage rates. Addition of pOSP during preincubation and fertilization significantly increased postcleavage development to blastocyst, but a synergistic stimulation on development was not detected when pOSP was included during in vitro culture. These results indicate that exposure to pOSP before and during fertilization reduces the incidence of polyspermy in pig oocytes, reduces the number of bound sperm, and increases postcleavage development to blastocyst.
First decision: 4 November 1999.
1 Supported by U.S. Department of Agriculture grant 97-35203-4614.
2 Correspondence: William C. Buhi, P.O. Box 100294, Department Obstetrics and Gynecology, University of Florida, Gainesville, FL 32610-0294. FAX: 352 392 2808; buhiwc{at}obgyn.ufl.edu
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