Effects of chilling to 0 degrees C on the morphology of meiotic spindles in human metaphase II oocytes.
Effects of chilling to 0 degrees C on the morphology of meiotic spindles in human metaphase II oocytes.
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DOI：
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发表时间：
2001
影响因子：
6.7
通讯作者：
M. Zenzes;R. Bielecki;R. Casper;S. Leibo
中科院分区：
文献类型：
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作者：
M. Zenzes;R. Bielecki;R. Casper;S. Leibo
目的通过光学切片显微镜观察0 ℃冷冻对人Ⅱ期卵母细胞纺锤体的影响。设计实验室研究。单位：一所医学院的学术研究实验室。患者（S）72名接受不孕症治疗的妇女共捐献了108个卵母细胞。干预（S）将中期II卵母细胞剥离其卵丘细胞，直接冷却至0 ℃，并保持1 - 10分钟。然后将它们固定在37 ℃下，进行免疫荧光染色，并进行显微镜检查。主要观察指标（S）冷冻和对照卵母细胞减数分裂纺锤体的形态。结果（S）与9个对照卵母细胞相比，46个冷冻卵母细胞的显微镜评价显示微管的各种时间依赖性变化。在0 ℃下1分钟后，纺锤体损伤可以忽略不计，但在冷却2或3分钟的卵母细胞中，纺锤体明显缩短，极性丧失。冷却到0摄氏度4到9分钟，导致越来越剧烈的变化;到10分钟时，纺锤体完全消失。尽管微管蛋白在0 ℃解聚，染色体没有分散，但即使在没有纺锤体的情况下仍保持锚定。结论（S）即使人类卵母细胞短暂暴露于接近0 ℃的温度也会引起减数分裂纺锤体的深刻改变。
OBJECTIVE To determine the effects of chilling to 0 degrees C on the meiotic spindle of human metaphase II oocytes, as observed by optical sectioning microscopy. DESIGN Laboratory study. SETTING Academic research laboratory in a medical school. PATIENT(S) Seventy-two women undergoing infertility treatment donated a total of 108 oocytes. INTERVENTION(S) Metaphase II oocytes were stripped of their cumulus cells, cooled directly to 0 degrees C, and held for periods of 1 to 10 minutes. They were then fixed at 37 degrees C, stained for immunofluorescence, and examined microscopically. MAIN OUTCOME MEASURE(S) Morphology of the meiotic spindle in chilled and control oocytes. RESULT(S) Microscopic evaluations of 46 chilled oocytes revealed various time-dependent changes in microtubules compared to 9 control oocytes. After 1 minute at 0 degrees C, spindle damage was negligible, but in oocytes cooled for 2 or 3 minutes, there was obvious shortening of the spindle and loss of polarity. Cooling to 0 degrees C for 4 to 9 minutes resulted in increasingly more drastic changes; by 10 minutes the spindles had totally disappeared. Despite depolymerization of microtubular tubulin at 0 degrees C, the chromosomes did not become dispersed, but remained anchored even in the absence of spindles. CONCLUSION(S) Even brief exposure of human oocytes to temperatures near 0 degrees C causes profound alterations of the meiotic spindle.