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Effect of Osmolality and Glycosaminoglycans on Motility, Capacitation, Acrosome Reaction, and In Vitro Fertilizability of Bovine Ejaculated Sperm¹

Department of Animal Science, University of Minnesota, St. Paul 55108

ABSTRACT

Bovine ejaculated semen was placed in a modified Tyrode's medium with albumin, lactate, and pyruvate. The sperm were washed three times and subjected to nine treatments in a 3 x 3 factorial arrangement. Treatments consisted of osmolality (exposure to 380 mOsmol/kg medium for 5 min, exposure to 340 or 295 mOsmol/kg medium for the entire incubation period), and the presence or absence of glycosaminoglycans (100 µg/ml chondroitin sulfate A or 10 µg/ml heparin). Sperm were examined at 4.5 h, 8 to 9 h, and 24 to 25 h of incubation (37°C, 5% CO₂> and 95% air). Heparin caused head-to-head agglutination of sperm, raised the percent sperm without seminal antigens over the acrosome (capacitated) by 20% at 4.5 h, and doubled the percent of acrosome-reacted sperm. However, this stimulation did not improve in vitro fertilizability. Chondroitin sulfate A tended to maintain motility, but did not affect capacitation or the acrosome reaction, possibly due to glucose inhibition. Both high osmolality treatments tended to reduce motility, especially after 24 h of incubation when the 340 osmolality treatment reduced motility by 14% over the 295 treatment. No consistent effect on capacitation was observed. The 340 and 380 osmolality treatments induced 8.6 and 6.1% more acrosome reactions by 24 h than the 295 treatment. The 340 mOsmol/kg treatment yielded insignificantly higher in vitro fertilization rates, as evidenced by development of zygotes to the two-cell stage. Lack of statistical significance was due to high variation with in vitro fertilization rates.

¹ Published as Paper Number 15007 of the Scientific Journal Series of the Minnesota Agricultural Experiment Station on research conducted under Minnesota Agricultural Experiment Station Project Number 72 supported by Hatch funds, the Graduate School, the University Computing Center, and Land O'Lakes, St. Paul.