RESUMO
BACKGROUND: Golden (Syrian) hamsters have many advantages for the study of reproductive biology and developmental biology, including a consistent estrous cycle, a stable superovulation response, and a short gestation period. However, there are serious difficulties in doing in vitro manipulations of hamster embryos, because they are very sensitive to various environmental factors. Thus, biotechnological researches of hamster embryos should be performed with high-level skills of embryo manipulations. METHODS: The authors summarized the history of hamster intracytoplasmic sperm injection (ICSI) and introduced key points for hamster ICSI, which were found in our previous studies on the production of embryos by ICSI and offspring by embryo transfer. MAIN FINDINGS: The key points for hamster ICSI were in vitro manipulations under the light-controlled environment, injection of acrosome-less sperm heads into oocytes as soon as possible before spontaneous oocyte activation occurs, and determination of the optimal culture conditions. CONCLUSION: To our knowledge, there are no available reports on production of offspring from ICSI embryos in hamsters except our reports. Moreover, success rates of hamster ICSI remain very low. For the purpose of spreading hamster ICSI, it is necessary to make further researches to improve manipulation techniques and to resolve experimental problems.
RESUMO
PURPOSE: To investigate the effects of sperm treatment medium-TCM199 or EGTA in Tris-HCl buffer (TBS + EGTA)-for sonication of frozen-thawed hamster spermatozoa in terms of sperm chromosome integrity and development of hamster oocytes injected with the sperm heads (ICSI). METHODS: Frozen-thawed hamster spermatozoa were separated into heads and tails by sonication in TCM199 or TBS + EGTA. Sperm heads were injected into mouse oocytes to assess hamster sperm chromosomes. We further compared the development of hamster ICSI embryos produced by injecting sonicated sperm heads in TCM199 vs TBS + EGTA. RESULTS: Sperm chromosome integrity was greater following sonication of frozen-thawed hamster spermatozoa in TBS + EGTA than in TCM199 (89.7% vs 69.0%). Embryonic development was improved following hamster oocyte injection with sperm heads sonicated in TBS + EGTA compared to in TCM199 (8-cell: 84.1% vs 65.4%; morula: 78.4% vs 43.2%; blastocyst: 42.0% vs 17.3%). Gene expression of zygotic genome activation in 2-cell embryos was significantly higher with TBS + EGTA than with TCM199. We transferred 43 morulae/blastocysts from the TBS + EGTA group to foster mothers, and 4 (9.3%) developed into live offspring. CONCLUSION: These results showed that the rapid injection of hamster sperm heads separated by sonication in TBS + EGTA effectively produced more ICSI embryos during a short time.
