Biotechnological studies on the mechanisms regulating embryogenesis

胚胎发生调控机制的生物技术研究

• 批准号: 05304021
• 负责人: TOYODA Yutaka
• 金额: $ 10.82万
• 依托单位: The University of Tokyo
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Co-operative Research (A)
• 财政年份: 1993
• 资助国家: 日本
• 起止时间: 1993 至 1994
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-05304021/
• 关键词: oocyte maturation; fertilization; c-mos; acrosin; endothelin; parthenogenesis; chimeras; embryo freezing; 遺伝子ターゲット法; c-mos遺伝子; 初期発生; 体外受精; 胚培養; 性比; 減数分裂; 卵丘; PCR

1)Oocyte maturation and fertilizationA)A heat-stable factor responsible for cumulus expansion was isolated form follicular fluid. The factor was potent to stimulate male pronucleus formation of eggs fertilized in vitro.B)Oocytes from c-mos-deficient mice matured to the second meiotic metaphase, but were activated without fertilization.C)Male mice homozygous for a targeted mutation in the mouse acrosin gene were still fertile.D)Supplementing the fertilization medium with soluble factor(s)vortexed out of cumulus-oocyte complexes improved the rate of fertilization when spermatozoa were preincubated in BSA-supplemented medium.2)Cleavage of fertilized eggsA)A steroid, progesterone, was potent to induce parthenogenetic development of unfertilized eggs.B)Fetal ovotestes in XX * XY chimeras develop into testes in adults.C)ET-1-'-homozygous mice die of respiratory failure at birth and have morphological abnormalities of the pharyngeal-arch-derived craniofacial tissues and organs.D)The cell line maintaining pluripotency was established in rat.3)Embryo freezingA)A system for producing young from cryopreserved gametes and embryos was established.

1)卵母细胞成熟和受精A）从卵泡液中分离出一种负责卵丘扩张的热稳定因子。该因子能有效刺激体外受精卵的雄性原核形成。B）来自c-mos缺陷小鼠的卵母细胞成熟至第二减数分裂中期，但在没有受精的情况下被激活。C）对于小鼠顶体酶基因中的靶向突变纯合的雄性小鼠仍然是可育的。D）用涡旋出卵丘的可溶性因子补充受精培养基。当精子在补充BSA的培养基中预孵育时，卵母细胞复合体提高了受精率。2）受精卵的卵裂A）类固醇，孕酮，B）XX * XY嵌合体中的胎儿卵睾丸发育成成年的睾丸。纯合子小鼠在出生时死于呼吸衰竭，并且具有咽弓衍生的颅面组织和器官的形态异常。D）在大鼠中建立了维持多能性的细胞系。3）胚胎冷冻A）建立了从冷冻保存的配子和胚胎产生幼仔的系统。

项目成果

Naito, K., Hawkins, C., Yamashita, M., Nagahama, Y., Aoki, F., Kohmoto, K., Toyoda, Y., Moor, R.M.: "Association of p34^<cdc2> and cyclin B1 during meiotic maturation in porcine oocytes." Dev.Biol.

Naito, K.、Hawkins, C.、Yamashita, M.、Nagahama, Y.、Aoki, F.、Kohmoto, K.、Toyoda, Y.、Moor, R.M.：“p34^<cdc2> 和细胞周期蛋白 B1 的关联

Aizaki, H., Watanabe, Y., Sarashina, T., Sakurai, T., Nagamori, S., Suzuki, T., Matsuura, Y., Saito, I., Miyamura, T.: "Genotyping of hepatitis C virus by simple Elisa method." Clin.Diagno.Virol.3. 105-109 (1994)

Aizaki, H.、Watanabe, Y.、Sarashina, T.、Sakurai, T.、Nagamori, S.、Suzuki, T.、Matsuura, Y.、Saito, I.、Miyamura, T.：“丙型肝炎的基因分型

馬場 忠: "An acrosomal protein,sp32,in mammalian sperm is a vinding protein specific for two proacrosins and an acrosin" Journal of Biological Chemistry. 269. 10133-10140 (1994)

Tadashi Baba：“哺乳动物精子中的顶体蛋白 sp32 是两种顶体蛋白原和顶体蛋白特异的结合蛋白”《生物化学杂志》269. 10133-10140 (1994)。

Baba, T., Niida, Y., Michikawa, Y., Kashiwabara, S., Kodaira, K., Takenaka, M., Kohno, N., Gerton, G.L., Arai, Y.: "An acrosomal protein, sp32, in mammalian sperm is a binding protein specific for two proacrosins and an acrosin intermediate." J.Biol.Chem.

Baba，T.，Niida，Y.，Michikawa，Y.，Kashiwabara，S.，Kodaira，K.，Takenaka，M.，Kohno，N.，Gerton，G.L.，Arai，Y.：“顶体蛋白，sp32

Baba, T., Azuma, S., Kashiwabara, S., Toyoda, Y.: "Sperm from mice carrying a targeted mutation of the acrosin gene can penetrate the oocyte zona pellucida and effect fertilization." J.Biol.Chem.269. 31845-31849 (1994)

Baba, T.、Azuma, S.、Kashiwabara, S.、Toyoda, Y.：“携带顶体蛋白基因靶向突变的小鼠的精子可以穿透卵母细胞透明带并实现受精。”