Induction of acrosome reactions of canine sperm by homologous zona pellucida

Abstract

In this study the induction of the acrosome reaction of canine sperm by homologous zona pellucida (ZP) was examined. Twelve semen samples obtained from 6 normal beagle dogs were evaluated after sperm incubation in vitro with canine capacitation medium (CCM). Washed sperm were preincubated at 37°C in 5% CO2 in air for 4 and 7 h prior to experimental treatment. Sperm were co- incubated for 1 min with intact oocytes collected from canine ovaries. Half of the oocytes were then fixed, and the bound sperm were assessed for acrosome reactions through use of a polyclonal antisperm antiserum and indirect immunofluorescence. The remaining oocytes were incubated in sperm- free medium for an additional 1-h period, and the acrosomal status of sperm bound to the ZP was evaluated similarly. The percentage of acrosome-reacted sperm on the ZP increased significantly during the 1-h incubation period. In other experiments, capacitated canine sperm were incubated with heat- solubilized ZP for 1 h and their acrosomal status was determined using fluoresceinated Pisum sativum lectin. The percentages of acrosome-reacted sperm increased significantly in ZP solution compared with controls. These data demonstrate that intact and solubilized canine ZP are capable of inducing acrosome reactions of canine sperm.