Development of a hamster superovulation program and adverse effects of gonadotropins on microfilament formation during oocyte development

Abstract

Objective: To establish a superovulation procedure for the golden hamster (Mesocricetus auratus) by elucidating gonadotropin effects on oocyte development. Design: Randomized, prospective study. Setting: University laboratory of embryology and gamete biotechnology. Animal(s): Twelve- to 15-week-old female and sexually mature male hamsters. Intervention(s): Different doses of pregnant mare serum gonadotropin (PMSG) were injected into female hamsters in metestrus, diestrus, or proestrus. The same dose of hCG was injected 56 hours later. Main Outcome Measure(s): Embryo development and oocyte morphology after treatment. Result(s): First, 10 IU or 15 IU each of PMSG and hCG was injected into 10 hamsters weighing 110 or 110–130 g, respectively. All hamsters were mated, but none delivered live young after injection. Second, the doses of 15 IU, 7.5 IU, 5 IU, or 0 IU of each gonadotropin were injected into each hamster (regardless of body weight, 5 per each group). Increasing numbers of embryos were retrieved as the dosage was increased (11.2 to 46.6 embryos per hamster), whereas the percentage of two-cell embryos at retrieval was significantly decreased (100% to 3%, P.05). In subsequent culture, none developed to blastocysts after 15-IU injection, whereas 47%, 55%, and 70% of two-cell embryos developed after 7.5-IU, 5-IU, and 0-IU treatments, respectively. As a result, females injected with 5 IU yielded more blastocysts than did females without injection (67 vs. 39). The number of inner cell mass cells per blastocyst was greatly increased in the control groups compared with the 5-IU and 7.5-IU treatment groups (22 vs. 14.3–14.7 cells per blastocyst). Third, the ultrastructure of oocytes was examined after injecting 5 IU each of PMSG and hCG (regardless of body weight). Superovulation did not affect oocyte maturation, but different patterns in microfilament formation were detected after the treatment. Conclusion(s): Female hamsters can be superovulated effectively by injecting equal amounts of PMSG and hCG, 56 hours apart. However, embryo development was adversely affected in a dose-dependent manner at all doses of gonadotropins, and microfilament distribution was affected by such treatment.