Effects of cooling germinal vesicle–stage bovine oocytes on meiotic spindle formation following in vitro maturation
Effects of cooling germinal vesicle–stage bovine oocytes on meiotic spindle formation following in vitro maturation
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冷却生发囊泡阶段牛卵母细胞对体外成熟后减数分裂纺锤体形成的影响
DOI：
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发表时间：
1999
影响因子：
2.5
通讯作者：
S. P. Leibo
中科院分区：
文献类型：
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作者：
Bin Wu;Jiangen Tong;S. P. Leibo
由于牛卵母细胞对接近0°C的温度敏感，低温保存牛卵母细胞的存活率很低。本研究评估了低温萌发囊泡期（GV）卵母细胞对微管和减数分裂纺锤体形成的影响。在实验1中，五组GV期卵母细胞，每组由~ 90个卵母细胞组成，分别在39°C保存作为对照，或在31°C保存，或在24、4或0°C冷却10分钟。处理后，所有卵母细胞在39℃下培养24小时。与对照组相比，将卵母细胞在31或24°C下保存10分钟并没有显著改变正常纺锤体的形成，但将它们冷却到4或0°C则会改变。成熟24小时后，在对照组或在31°C或24°C保存的卵母细胞中含有正常减数分裂纺锤体的比例分别为69.8%、71.9%和69.4% （P < 0.05）。相比之下，卵母细胞在冷却至4°C或0°C后，纺锤体正常的百分比分别为44.0%和29.1%。在实验2中，将~ 90个卵母细胞/组冷却至4°C多次，然后加热培养。无论暴露时间如何，将卵母细胞冷却到4°C都会减少正常纺锤体的形成。卵母细胞在4℃下冷却10、20、30、45、60 min后纺锤体正常的比例分别为44.0%、38.4%、37.5%、34.5%和30.9%。在实验3中，每组约60个卵母细胞在31°C下保存或冷却至24、4或0°C 10分钟，使其成熟24小时，然后进行体外受精。不同低温处理的卵母细胞的卵裂率与正常纺锤体卵母细胞的卵裂率相同。也就是说，对照卵母细胞与放置在31°C或24°C的卵母细胞的卵裂率无显著差异（70.4%、71.8%和72.4%;P < 0.05）。然而，冷冻至4°C和0°C的卵母细胞在受精后分裂的比例分别为37.0%和30.4%。以上结果表明：(1)低温处理牛卵母细胞不低于24℃，不会减少正常减数分裂纺锤体的形成；(2)然而，将卵母细胞冷却至4°C或更低温度仅10分钟，就会大大减少正常减数分裂纺锤体的形成和受精；(3)受精率和最终受精卵的卵裂率与正常纺锤体的形成率相似。摩尔。天线转换开关。发展。54:38 - 395,1999。©1999 Wiley‐Liss, Inc。
Attempts to cryopreserve bovine oocytes result in low survival because of their sensitivity to temperatures near 0°C. This study evaluates the effects of chilling germinal vesicle‐stage (GV) oocytes on their formation of microtubules and the meiotic spindle. In experiment 1, five groups of GV‐stage oocytes, each consisting of ∼90 oocytes, were held at 39°C as controls, or at 31°C, or cooled to 24, 4 or 0°C for 10 min. After being treated, all oocytes were cultured at 39°C for 24 hr. Compared to the controls, holding oocytes for 10 min at 31 or 24°C did not significantly alter the formation of normal spindles, but chilling them to 4 or 0°C did. After 24 hr of maturation, the respective percentages of oocytes containing normal meiotic spindles observed in the controls or those held at 31 or 24°C were 69.8%, 71.9%, or 69.4% (P > 0.05). In contrast, the percentages of oocytes with normal spindles after they had been cooled to 4 or 0°C were 44.0% or 29.1%, respectively. In experiment 2, ∼90 oocytes/group were cooled to 4°C for various times before being warmed and cultured. Regardless of the time of exposure, cooling oocytes to 4°C reduced the formation of normal spindles. The percentages of oocytes cooled to 4°C for 10, 20, 30, 45, or 60 min with normal spindles were 44.0%, 38.4%, 37.5%, 34.5% and 30.9%, respectively. In experiment 3, ∼60 oocytes per group that had been held at 31°C or cooled to 24, 4 or 0°C for 10 min were allowed to mature for 24 hr before being subjected to in vitro fertilization. The cleavage rates of oocytes subjected to various chilling treatments exhibited the same pattern as that of oocytes with normal spindles. That is, there were no significant differences in cleavage rates among the control oocytes and those held at 31 or 24°C (70.4%, 71.8%, and 72.4%; P > 0.05). However, only 37.0% and 30.4% of oocytes chilled to 4 or 0°C cleaved after fertilization. These results suggest that: (1) chilling bovine oocytes no lower than 24°C does not reduce formation of normal meiotic spindles; (2) however, chilling oocytes to 4°C or lower for as little as 10 min drastically reduces the formation of normal meiotic spindles and of fertilization; (3) the rates of fertilization and cleavage of resultant zygotes mimic that of formation of normal spindles. Mol. Reprod. Dev. 54:388–395, 1999. © 1999 Wiley‐Liss, Inc.
影响因子：
3.6
作者：
Bin Wu;G. Ignotz;W. Bruce Currie;Xiangzhong Yang
通讯作者：
Bin Wu;G. Ignotz;W. Bruce Currie;Xiangzhong Yang
影响因子：
2.7
作者：
EPPIG, JJ;SCHULTZ, RM;CHESNEL, F
通讯作者：
CHESNEL, F
影响因子：
3.6
作者：
AMAN, RR;PARKS, JE
通讯作者：
PARKS, JE