Removal of paternal male accessory sex glands (ASG) could cause a delay in DNA synthesis in hamster zygotes fertilized in vivo. In view of the fact that this process is closely related to pronuclear development which, in part, depends on sperm nuclear decondensation and oocyte activation during fertilization, we carried out a series of experiments were undertaken to determine whether ASG also has an effect on these early events. (1) Oocytes were collected from females mated with SH (sham-operated control), AGX (bilateral excision of ampullary glands), VPX (bilateral excision of ventral prostates) or TX (excision of all ASG) males (n = 8 per group) at 4, 5 and 6 h post coitus. (2) Epididymal spermatozoa were incubated with total ventral prostate (VP) secretion to study its effect on dithiothreitol-induced sperm decondensation. (3) Histone H1 kinase activity in oocytes collected as described in (1) was determined. (4) Exocytosed cortical granules on oocytes were labelled with FITC-LCA and quantified by a Metamorph Imaging System. Results showed that sperm decondensation and resumption of meiosis in oocytes in VPX and TX groups were significantly slower compared with SH. VP secretion augmented sperm decondensation in vitro. At 4 h post coitus, the relative activity of histone H1 kinase in the TX and VPX groups was significantly higher than that in the SH group (p < 0.01). Cortical granule exocytosis in the AGX group was consistently weaker at all time points studied and was significantly lower than that of the control at 4 h post coitus (p < 0.05), while the percentage of polyspermic fertilization in the AGX group was significantly higher compared with that in the SH group (p < 0.05). Taken together, these results show that the lack of exposure of spermatozoa to secretions of the ASG does not jeopardize their ability to penetrate ova, although other aspects of their function in the early stages of gamete interaction and subsequent initiation of embryonic development are affected.