Developmental Ability of Vitrified Mouse Oocytes Expressing Water Channels
| Autor: | Kazutsugu Matsukawa, Keisuke Edashige, Shinsuke Seki, Magosaburo Kasai, Yohei Yamaji, Chihiro Koshimoto |
|---|---|
| Rok vydání: | 2011 |
| Předmět: |
Glycerol
Membrane permeability Cryoprotectant Fertilization in Vitro Biology General Biochemistry Genetics and Molecular Biology Andrology Mice Cryoprotective Agents Human fertilization medicine Animals Zona pellucida Zona Pellucida Aquaporin 3 Mice Inbred ICR Cumulus Cells Germinal vesicle Zygote urogenital system General Medicine Oocyte Vitrification Cell biology Blastocyst medicine.anatomical_structure Immunology Oocytes Female Animal Science and Zoology General Agricultural and Biological Sciences |
| Zdroj: | Journal of Reproduction and Development. 57:403-408 |
| ISSN: | 1348-4400 0916-8818 |
| DOI: | 10.1262/jrd.10-201m |
| Popis: | Previously, we showed that the exogenous expression of aquaporin 3 (AQP3), an aquaglyceroporin, improved the tolerance of mouse oocytes to vitrification with a glycerol-based solution. In the present study, we examined conditions suitable for the expression of AQP3 and the ability of vitrified oocytes to develop in vitro and in vivo after fertilization. After only partial remove of cumulus cells, immature mouse oocytes (germinal vesicle stage) were injected with 5, 10 or 20 pg of AQP3 cRNA and cultured for 12 h for maturation. When matured oocytes were vitrified with a glycerol-based solution, 57-61% survived regardless of the amount of cRNA injected (5-20 pg). By contrast, no oocytes injected with water (control) survived. When the zona pellucida was removed from the vitrified oocytes and the oocytes were then fertilized in vitro and cultured, the proportions that were fertilized and developed into blastocysts were higher when the amount of cRNA injected was 5 pg than 10-20 pg. When 16 blastocysts were transferred to a pseudopregnant mouse, 5 developed to term, demonstrating that oocytes vitrified after injection of AQP3 cRNA retained the ability to develop to term. The water-permeability of cRNA-injected oocytes was higher than that of control oocytes from the maturing stage to the 1-cell zygote stage, whereas glycerol-permeability was higher only at metaphase II. This indicates that AQP3 was expressed for a relatively short period of time. These results suggest that the transient expression of water/cryoprotectant channels is effective for cryopreserving cells that have low membrane-permeability, such as mammalian oocytes. |
| Databáze: | OpenAIRE |
| Externí odkaz: |
|