Role of cell surface carbohydrates on the development of mammalian embryos in vitro

细胞表面碳水化合物对哺乳动物胚胎体外发育的作用

• 批准号: 05660327
• 负责人: YAMADA Masayasu
• 金额: $ 1.34万
• 依托单位: KYOTO UNIVERSITY
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for General Scientific Research (C)
• 财政年份: 1993
• 资助国家: 日本
• 起止时间: 1993 至 1995
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/en/grant/KAKENHI-PROJECT-05660327/
• 关键词: preimplantation; embryo; in vitro development; PHA-P lectin; compaction; N-linled carbohydrates; oviduct extracts; ラット; 発生; PHAレクチン; 卵管因子; マウス; 糖鎖; Nグリカナーゼ

The developmental block at the 2-cell stage in rat embryos cultured in vitro has recently been overcome by deleting phosphate and glucose from culture medium. However, the problem was still remained that early 1-cell embryos were less able to develop in vitro to the blastocyst stage than 2-cell embryos. To improve the culture conditions for the 1-cell embryos, we first examined the effect of PHA-P lectin, which is known to induce in vitro maturation of mouse oocytes. As the results, it was found that PHA-P,especially E_4 subunit promoted the developmental ability of rat 1-cell embryos to blastocysts in vitro, suggesting that the action of PHA-P is specific and a modification of cell surface carbohydrate moieties of the embryos. Furthermore, we found that development of rat 1-cell embryos to blastocysts was promoted with macromolecules in rat oviduct extracts, whose molecular mass was about 70kDa.Concerning the study on the involvement of Cell surface carbohydrates in compaction in the mouse embryos at the morula stage, it was revealed that cell surface N-linked carbohydrates that were specifically cleaved by N-glycanase participated greatly in the compaction process.

体外培养的大鼠胚胎在2-细胞期的发育阻滞最近已通过从培养基中去除磷酸盐和葡萄糖而被克服。然而，问题仍然存在，早期1-细胞胚胎在体外发育到囊胚阶段的能力低于2-细胞胚胎。为了改善1-细胞胚胎的培养条件，我们首先检测了已知诱导小鼠卵母细胞体外成熟的PHA-P凝集素的作用。结果表明，PHA-P，尤其是E_4亚基能促进大鼠1-细胞胚胎体外发育，提示PHA-P的作用是特异性的，是对胚胎细胞表面糖基的修饰。此外，我们发现大鼠输卵管提取物中分子量约为70 kDa的大分子物质能够促进大鼠1-细胞胚胎向囊胚的发育。关于细胞表面碳水化合物参与小鼠桑椹胚期胚胎致密化过程的研究，发现被N-聚糖酶特异性切割的细胞表面N-连接碳水化合物参与了致密化过程。

项目成果

Hayashi, H.: "Effect of cell surface N-linked carbohydrates on the compaction of preimplantation mouse embryos" J.Soc.Immunol.Reprod.88-92 (1994)

Hayashi, H.：“细胞表面 N 连接碳水化合物对植入前小鼠胚胎压缩的影响”J.Soc.Immunol.Reprod.88-92 (1994)

Y.HOSOI: "Isolation and characterization of cell lines derived from rat embryos cultured in vitro." Jpn.J.Embryo Transfer. 17. 13-18 (1995)

Y.HOSOI：“体外培养的大鼠胚胎细胞系的分离和表征。”

Minami, N.: "Nonspecies-specific effects of mouse oviducts on the development of bovine IVM/IVF embryos by a serum free co-culture." Theriogenology. 41. 1435-1445 (1994)

Minami, N.：“通过无血清共培养，小鼠输卵管对牛 IVM/IVF 胚胎发育的非物种特异性影响。”

Yamada, M.: "Effects of Nacl and KH_2PO_4 on in vitro development of 1-cell rat embryos in a protein-free defined medium." J. Assist, Reprod. Genet.10. 164 (1993)

Yamada, M.：“Nacl 和 KH_2PO_4 对无蛋白确定培养基中 1 细胞大鼠胚胎体外发育的影响。”

Kim, K. S.: "The effects of follicular fluid on in vitro maturation, fertilization and the development of bovine embryos." Theriogenology. 45. 787-799 (1996)

Kim, K. S.：“卵泡液对牛胚胎体外成熟、受精和发育的影响。”