Establishment of cryo-preservation method for ovary and testis and storage of wild mouse genes.

卵巢、睾丸冷冻保存方法的建立及野生小鼠基因的保存。

• 批准号: 08680918
• 负责人: TAYA Choji
• 金额: $ 1.15万
• 依托单位: THE TOKYO METROPOLITAN INSTITUTE OF MEDICAL SCIENCE
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (C)
• 财政年份: 1996
• 资助国家: 日本
• 起止时间: 1996 至 1997
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-08680918/
• 关键词: Cryo-preservation of ovary; Cryo-preservation of testis; Wild mouse genes; Preservation of gene resources

This study aimed at an establishment of storage system for wild mouse genome by freezing of ovary and testis from a viewpoint of preservation of gene resources.1. Establishment of cryo-preservation method for ovary and strage of wild mouse genes.Successful freezing conditions of ovaries were detected in a series of experiments using laboratory mice. When the freesing method was applied to ovaries of MSM inbred strain derived from Japanese wild mice, mice derived from frozen ovaries were obtained, though litter sizes were very small.2. Establishment of cryo-preservation method for testisfreezing injuries of newborn mouse testes were examined histologically at various points of a freezing course. A lot of germ cells retained a normal morphology in the program of slow cooling til -40ﾟC before immersion into LN2.In order to confirm the growth of germ cells and the production of spermatozoa, the frozen newborn testis were transplanted into histocompatible adult testis. In the case of a freezing program described above, at 14 days after transplantation meiotic germ cells were observed and at six weeks spermatozoa were detected in the seminiferous tubules derived from the frozen testes, though it has not been confirmed whether they have a capacity of fertilization to oocytes.A reconstitution of thawed testis to a host reproductive system is under investigation.

本研究旨在从基因资源保存的角度出发，建立野生小鼠卵巢和睾丸冷冻保存基因组的方法.卵巢冷冻保存方法的建立及野生小鼠基因片段的扩增利用实验小鼠进行卵巢冷冻保存的实验研究。对日本野生小鼠来源的MSM近交系小鼠进行卵巢解冻，获得了冷冻卵巢小鼠，但胎仔数很少.睾丸冷冻保存方法的建立对冷冻过程中不同时间点的新生小鼠睾丸冷冻损伤进行了组织学观察。冷冻至-40 ℃后，液氮浸泡，大量生殖细胞仍保持正常形态。为了证实生殖细胞的生长和精子的产生，将冷冻的新生睾丸移植到组织相容性的成年睾丸中。在上述冷冻程序的情况下，在移植后14天观察到减数分裂的生殖细胞，并且在6周时在来自冷冻睾丸的生精小管中检测到精子，尽管尚未证实它们是否具有受精成卵母细胞的能力。

项目成果

Shimmoto, M.: "Generation and charactrzaton of transgenic mice expressing a human mutant d-galactosidase with af R301Q substitution causing a variant form of Fabry disease." FEBS Lett.417. 89-91 (1997)

Shimmoto, M.：“表达带有 af R301Q 取代的人类突变 d-半乳糖苷酶的转基因小鼠的产生和特征，导致法布里病的变异形式。”

Shimmoto,M.: "Generation and characterization of transgenic mice expressing a human mutant α-galactosidase with an R301Q substitution causing a variant form of Fabry disease." FEBS Lett.417. 89-91 (1997)

Shimmoto, M.：“表达具有 R301Q 取代的人类突变 α-半乳糖苷酶的转基因小鼠的生成和表征，导致法布里病的变异形式。”417 (1997)。

Ishii S.: "α-Galactosidase transgenic mouse:Heterogenous gene expression and posttranslational glycosylation." Glycoconjugate J.(in press). (1997)

Ishii S.：“α-半乳糖苷酶转基因小鼠：异质基因表达和翻译后糖基化。”（出版中）。

Ishii, S.et al.: "Alpha-garactosidase transgenic mouse : Heterogenous gene expression and posttranslational glycosylation." Glycoconjugate J.(in press). (1997)

Ishii, S.et al.：“α-半乳糖苷酶转基因小鼠：异源基因表达和翻译后糖基化。”

Shimmoto, M.et al: "Generation and characterization of transgenic mice expressing a human mutant alpha-galactosidase with an R301Q substitution causing a variant form of Fabry disease." FEBS Lett.417. 89-91 (1997)

Shimmoto, M.等人：“表达人类突变型 α-半乳糖苷酶的转基因小鼠的生成和表征，该酶具有 R301Q 取代，导致法布里病的变异形式。”