Biology of Reproduction, Vol 53, 1280-1285, Copyright © 1995 by Society for the Study of Reproduction

ARTICLES

Sperm motility hyperactivation facilitates penetration of the hamster zona pellucida

CR Stauss, TJ Votta and SS Suarez
Department of Physiological Science, College of Veterinary Medicine, University of Florida, Gainesville 32610-0144, USA.

These experiments were conducted to determine whether or not sperm motility hyperactivation facilities penetration of the zona pellucida of the oocyte. Two approaches were used. For the first, hamster sperm were incubated for 4.0-4.25 h in a capacitating medium that contained either 2.9 or 25.0 mM sodium bicarbonate. In these media, sperm became equally capacitated as evidenced by their ability to undergo the acrosome reaction when exposed to lysophosphatidyl choline or intact zonae pellucidae; however, sperm became hyperactivated only in the medium containing 25.0 mM bicarbonate. When these sperm were added to cumulus-free oocytes in vitro, only 2 of 88 oocytes were penetrated by sperm preincubated in 2.9 mM bicarbonate, while 31 of 86 oocytes were penetrated by sperm in 25.0 mM bicarbonate. It was found that equal numbers of sperm were bound to the oocytes and that equal numbers were acrosome-reacted on the surface of the zonae in the two media. For the second approach, sperm were incubated in a capacitating medium containing 25 mM bicarbonate. When > 70% were hyperactivated, aliquots were added to three sets of oocytes. After 10 min had been allowed for sperm to attach and acrosome-react, inhibitors of hyperactivation were added and the sperm and oocytes were incubated for an additional 20 min before fixation and examination for zona penetration. In the dishes treated with the inhibitors verapamil or Cd2+, 1 of 42 and 0 of 42 oocytes were penetrated, respectively, compared with 25 of 40 in controls. Therefore, it appears that hyperactivation facilitates penetration of the hamster zona pellucida.