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1,2 - 丙二醇与1,2 - 乙二醇对家猫后续卵母细胞成熟、纺锤体完整性、受精及体外胚胎发育的影响

Effect of 1,2-propanediol versus 1,2-ethanediol on subsequent oocyte maturation, spindle integrity, fertilization, and embryo development in vitro in the domestic cat.

作者信息

Comizzoli Pierre, Wildt David E, Pukazhenthi Budhan S

机构信息

Department of Reproductive Sciences, Smithsonian's National Zoological Park, 3001 Connecticut Avenue NW, Washington, D.C. 20008, USA.

出版信息

Biol Reprod. 2004 Aug;71(2):598-604. doi: 10.1095/biolreprod.104.027920. Epub 2004 Apr 14.

Abstract

This study assessed the impact of various cryoprotectant (CPA) exposures on nuclear and cytoplasmic maturation in the immature cat oocyte as a prerequisite to formulating a successful cryopreservation protocol. In experiment 1, immature oocytes were exposed to 0, 0.75, 1.5, or 3.0 M of 1,2-propanediol (PrOH) or 1,2-ethanediol (EG) at room temperature (25 degrees C) or 0 degrees C for 30 min. After CPA removal and in vitro maturation, percentage of oocytes reaching metaphase II (MII) was reduced after exposure to 3.0 M PrOH at 0 degrees C or 3.0 M EG at both temperatures. All CPA exposures increased MII spindle abnormalities compared to control, except 1.5 M PrOH at 25 degrees C. In experiments 2 and 3, immature oocytes were exposed to CPA conditions yielding optimal nuclear maturation that either had caused spindle damage (0.75 M PrOH, 1.5 M EG, and 3.0 M PrOH at 25 degrees C) or not (1.5 M PrOH at 25 degrees C). After maturation and insemination in vitro, oocytes were cultured for 7 days to assess treatment influence on developmental competence. CPA exposure did not affect fertilization, but the high incidence of MII spindle abnormalities resulted in a low percentage of cleaved embryos. Blastocyst formation and quality were influenced by both CPA types (EG was more detrimental than PrOH) and concentration (3.0 M was more detrimental than 1.5 M). Overall, cat oocytes appear to be highly sensitive to CPA except after exposure to 1.5 M PrOH at 25 degrees C, a treatment that still allowed approximately 60% of the oocytes to reach MII and approximately 20% to form blastocysts.

摘要

本研究评估了各种冷冻保护剂(CPA)处理对未成熟猫卵母细胞核成熟和细胞质成熟的影响,以此作为制定成功冷冻保存方案的前提条件。在实验1中,未成熟卵母细胞在室温(25℃)或0℃下,分别暴露于0、0.75、1.5或3.0M的1,2 - 丙二醇(PrOH)或1,2 - 乙二醇(EG)中30分钟。去除CPA并进行体外成熟培养后,在0℃下暴露于3.0M PrOH或在两个温度下暴露于3.0M EG后,达到中期II(MII)的卵母细胞百分比降低。与对照组相比,所有CPA处理均增加了MII纺锤体异常,25℃下1.5M PrOH处理除外。在实验2和3中,未成熟卵母细胞暴露于能产生最佳核成熟的CPA条件下,这些条件有的会导致纺锤体损伤(25℃下0.75M PrOH、1.5M EG和3.0M PrOH),有的则不会(25℃下1.5M PrOH)。体外成熟和授精后,将卵母细胞培养7天以评估处理对发育能力的影响。CPA处理不影响受精,但MII纺锤体异常的高发生率导致分裂胚胎的百分比很低。囊胚形成和质量受到CPA类型(EG比PrOH更有害)和浓度(3.0M比1.5M更有害)的影响。总体而言,猫卵母细胞似乎对CPA高度敏感,25℃下暴露于1.5M PrOH除外,该处理仍能使约60%的卵母细胞达到MII,约20%形成囊胚。

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