The regulation of early phase development of mammalian embryos

哺乳动物胚胎早期发育的调控

• 批准号: 03640620
• 负责人: NAKAMURA Masahisa
• 金额: $ 1.34万
• 依托单位: Teikyo Junior College
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for General Scientific Research (C)
• 财政年份: 1991
• 资助国家: 日本
• 起止时间: 1991 至 1992
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-03640620/
• 关键词: Mouse; Two-cell block; ATP; NaF; Glycolysis; 2-cell block; 初期胚; 2ーcell block

This study was undertaken to elucidate the mechanism underlying the inability of mouse embryos to develop in culture past the 2-cell stage (2-cell block). Sodium fluoride (NaF, an inhibitor of enolase in glycolysis) significantly improved the rate of further development of 2-cell embryos in a dose-fashion manner. The removal of glucose and phosphate from the conventional medium improved the rate of development of 2-cell embryos to* 3 cell (42%), but the rate of development of 2-cell embryos to* 3 cells was enhanced (77%) when NaF at 2.5 mM was added to the medium without glucose and phosphate. The effect of NaF at 2.5 mM on ^<14>CO_2 and [^<14>C] lactate production form [^<14>C] glucose was also examined. NaF at 2.5 mM significantly inhibited both ^<14>CO_2 and [^<14>C] lactate production from [6-^<14>C] glucose, but not ^<14>CO_2 production from [1-^<14>C]-glucose, although it had no effect on the uptake of [^3H] deoxy-D-glucose (a non-metabolizable analogue of glucose) by the embryos. When the intracellular level of adenosine triphosphate (ATP) during early development of mouse embryos was examined, it was higher before rather than after cleavage. In addition, the ATP level was much higher before the 2nd cleavage 30 hr after insemination in unblocked embryos than that in blocked ones. These results suggest that the flow rate of glucose in glycolysis may regulate the development of 2-cell embryos to*3 cells, and that the intracellular level of ATP may be critical to early phase of mouse embryo development in vitro.

这项研究是为了阐明小鼠胚胎不能在超过2-细胞阶段(2-细胞块)的培养中发育的机制。氟化钠(NaF，糖酵解中烯醇化酶的抑制剂)以剂量-方式显著提高2-细胞胚胎的进一步发育率。常规培养液中去除葡萄糖和磷酸盐可提高2-细胞胚胎向*3细胞的发育率(42%)，但添加2.5 mM的NaF可提高2-细胞胚胎向*3细胞的发育率(77%)。此外，还考察了2.5 mM NaF对~(14&t；14&gt；CO_2)和[~(14&lt；14&gt；C)]葡萄糖生成的影响。2.5 mM的NaF显著抑制[6-^lt；14&gt；C]葡萄糖产生乳酸和[^&lt；14&gt；C]乳酸，但对[1-^lt；14&gt；C]-葡萄糖产生^&lt；14&gt；CO_2无明显影响，但对胚胎摄取脱氧-D-葡萄糖(葡萄糖的一种非代谢类似物)没有影响。检测小鼠胚胎早期发育过程中细胞内三磷酸腺苷(ATP)水平时，卵裂前高于卵裂后。此外，未阻断胚胎在受精后30小时第二次卵裂前的ATP水平明显高于阻断胚胎。这些结果表明，葡萄糖在糖酵解中的流动速度可能调节2-细胞胚胎向*3细胞的发育，细胞内ATP的水平可能是小鼠胚胎体外发育早期的关键。

项目成果

Nakamura et al.: "Purification and localization of a Ca^<2+>-binding protein in human spermatozoa." Human Reproduction. 7. 500-505 (1992)

Nakamura 等人：“人类精子中 Ca^2 结合蛋白的纯化和定位。”

NAKAMURA Etal.: "Calretiwliu is present in the a crosome of spermatiels of rot testis" Biochew Biophys Res Commun. 186. 668-673 (1992)

NAKAMURA 等人：“Calretiwliu 存在于腐烂睾丸的精子顶体中”Biochew Biophys Res Commun。

NAKAMURA Etal.: "Artisera to calretiwlin in hibits speron motility in mice" Biochem Biphys.Res commun.186. 984-990 (1992)

NAKAMURA 等人：“Artisera 至 calretiwlin 抑制小鼠 speron 运动”Biochem Biphys.Res commun.186。

Masahisa Nakamura et al: "Purification and localization of a calcium- binding protein in human spermatozoa" Human Reproduction. 7. 500-505 (1992)

Masahisa Nakamura 等人：“人类精子中钙结合蛋白的纯化和定位”人类生殖。

NAKAMURA Etal.: "Purificotion and locoligation of a Ca_<2+>-binding protein in huwan sperwatigoa." Human Reproduction. 7. 500-505 (1992)

NAKAMURA 等人：“Huwan sperwatigoa 中 Ca_2 结合蛋白的纯化和定位”。