Morató Roser, Mogas Teresa, Maddox-Hyttel Poul
Departament de Medicina i Cirurgia Animals. Facultat de Veterinària. Universitat Autònoma de Barcelona, Bellaterra, Spain.
Mol Reprod Dev. 2008 Aug;75(8):1318-26. doi: 10.1002/mrd.20873.
Our objective was to document potential subcellular consequences of treatment with the microtubule stabilizer Taxol with or without subsequent vitrification of cow and calf oocytes by the open pulled straw (OPS) method. Oocytes were divided into four experimental groups for cows and four groups for calves: (1) a control group fixed immediately after maturation; (2) an OPS group cryopreserved by conventional OPS; (3) a Taxol/CPA group exposed to 1 microM Taxol and cryoprotective agents (CPAs); and (4) a Taxol/OPS group vitrified by OPS including 1 microM Taxol to the vitrification solution. All oocytes were processed for light and transmission electron microscopy. The main injuries were observed on the metaphase plate and the spindle. In control oocytes, the metaphase appeared as condensed chromosomes arranged in a well-organized metaphase plate and the spindle showed well organized microtubules in both cow and calf oocytes. However, in cow OPS oocytes, the metaphase plate was disorganized into scattered chromosomes or the chromosomes were condensed into a single block of chromatin. In addition, microtubules were not organized as typical spindles. In contrast, cow Taxol/OPS oocytes as well as both cow and calf Taxol/CPAs oocytes showed well-organized metaphase plates and normal spindle morphology. All calf OPS and calf Taxol/OPS oocytes displayed a single block of chromatin and no microtubules could be observed around the chromosomes. In conclusion, treatment with 1 microM Taxol before and during vitrification did not induce adverse changes in the oocyte cytoplasm or metaphase spindles in adult bovine oocytes, but stabilized the metaphase and spindle morphology.
我们的目标是记录用微管稳定剂紫杉醇处理牛和小牛卵母细胞后，无论随后是否通过开放式拉长细管（OPS）法进行玻璃化处理，其潜在的亚细胞后果。卵母细胞被分为牛的四个实验组和小牛的四个实验组：（1）成熟后立即固定的对照组；（2）通过传统OPS法冷冻保存的OPS组；（3）暴露于1微摩尔紫杉醇和冷冻保护剂（CPA）的紫杉醇/CPA组；（4）通过包含1微摩尔紫杉醇的OPS法玻璃化处理至玻璃化溶液的紫杉醇/OPS组。所有卵母细胞均进行光镜和透射电镜处理。主要损伤出现在中期板和纺锤体上。在对照卵母细胞中，中期表现为浓缩染色体排列在组织良好的中期板上，并且纺锤体在牛和小牛卵母细胞中均显示出组织良好的微管。然而，在牛的OPS卵母细胞中，中期板紊乱成散在的染色体，或者染色体浓缩成单个染色质块。此外，微管没有组织成典型的纺锤体。相比之下，牛的紫杉醇/OPS卵母细胞以及牛和小牛的紫杉醇/CPA卵母细胞均显示出组织良好的中期板和正常的纺锤体形态。所有小牛的OPS和小牛的紫杉醇/OPS卵母细胞均显示单个染色质块，并且在染色体周围未观察到微管。总之，在玻璃化处理之前和期间用1微摩尔紫杉醇处理不会在成年牛卵母细胞的卵母细胞质或中期纺锤体中诱导不良变化，而是稳定了中期和纺锤体形态。
