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Published on: March 1, 2014
Development of Mongolian gerbil embryos in chemically defined media: effects of osmolarity, glucose and phosphate
Hirotada Tsujii1, Naoko Taniguchi, Koh-ichi Hamano
1Department of Agricultural Biotechnology, Faculty of Agriculture, Shinshu University, Kamiina, Japan. htsujii@gipmc.shinshu-u.ac.jp
Abstract:
Mongolian gerbil 2-cell embryos were cultured in modified M16. When osmolarity of the medium with 5.0 mmol glucose l(-1) was varied by adjusting the amount of NaCl added, 2-cell embryos at 280, 290, 300 and 310 mOsmol developed to the 8- and 16-cell stages. The incorporation and oxidation of 14C-Methionine were compared between fresh recovered and cultured embryos at the 1-cell to 16-cell stages. Development beyond the 8-cell stage of fresh recovered embryos showed an enhanced rate of total protein synthesis, indicating activation of the transcription process of the embryonic genome. However, we found that the lowest incorporation and oxidation of 14C-Methionine was observed in cultured embryos of the 16-cell stage at 115 h after hCG injection. In the medium without phosphate, glucose promoted development of 2-cell embryos to the 8-cell stage, and low concentrations of glucose were necessary for the development of the 2-cell to 8-cell stages. These results suggest that Mongolian gerbil preimplantation embryos can be cultured in vitro in a chemically defined medium with a low concentration of glucose.
Insights
Mongolian gerbil embryos can develop in vitro using a defined medium with low glucose. Optimal osmolarity supports development to the 16-cell stage, though protein synthesis differs from fresh embryos.
Area of Science:
- Developmental Biology
- Embryology
- Reproductive Science
Background:
- Preimplantation embryo development is crucial for successful reproduction.
- Understanding optimal culture conditions is essential for assisted reproductive technologies.
- Mammalian embryo development relies on specific nutrient and osmotic balances.
Purpose of the Study:
- To investigate the effects of osmolarity and glucose concentration on Mongolian gerbil embryo development in vitro.
- To compare metabolic activity (methionine incorporation and oxidation) between in vitro cultured and fresh embryos.
- To determine the feasibility of culturing gerbil embryos in a chemically defined medium.
Main Methods:
- Culturing Mongolian gerbil 2-cell embryos in modified M16 medium with varying osmolarity (280-310 mOsmol) and glucose concentrations.
- Assessing embryo development to the 8- and 16-cell stages.
- Measuring 14C-methionine incorporation and oxidation in embryos from 1-cell to 16-cell stages.
- Evaluating embryo development in a phosphate-free medium with varying glucose levels.
Main Results:
- Embryos developed to the 8- and 16-cell stages across a range of osmolarities (280-310 mOsmol) with 5.0 mmol/L glucose.
- Freshly recovered embryos showed enhanced protein synthesis beyond the 8-cell stage, indicating genome activation.
- Cultured 16-cell embryos exhibited lower methionine incorporation and oxidation compared to fresh embryos.
- Glucose promoted 2-cell to 8-cell development in phosphate-free medium; low glucose concentrations were necessary.
Conclusions:
- Mongolian gerbil preimplantation embryos can be successfully cultured in vitro.
- A chemically defined medium with low glucose concentrations is suitable for gerbil embryo development.
- Osmolarity plays a role in supporting early embryonic development in vitro.
- Metabolic differences exist between in vitro cultured and in vivo developed embryos.
