EFFECTS OF COOLING AND REWARMING ON THE MEIOTIC SPINDLE AND CHROMOSOMES OF IN VITRO-MATURED BOVINE OOCYTES
EFFECTS OF COOLING AND REWARMING ON THE MEIOTIC SPINDLE AND CHROMOSOMES OF IN VITRO-MATURED BOVINE OOCYTES
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DOI：
10.1095/biolreprod50.1.103
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发表时间：
1994-01-01
影响因子：
3.6
通讯作者：
PARKS, JE
中科院分区：
文献类型：
--
作者：
AMAN, RR;PARKS, JE
本研究旨在探讨降温和复温对牛卵母细胞减数分裂纺锤体的影响。在实验1中，牛卵母细胞体外成熟后要么保持在39%，要么突然冷却到4℃或接近25℃。免疫组织化学和DNA染色分别显示牛卵母细胞内有倒星形的桶形纺锤形微管和染色体。在4℃下暴露10-20分钟，纺锤体完全消失。在4℃恒温处理30min后，90%的卵母细胞出现异常，出现纺锤体异常或无纺锤体。在实验2中，将卵母细胞冷却至类似于25℃或4℃的温度30分钟，然后直接或分步复温15或60分钟。无论降温或复温方案如何，大多数卵母细胞的纺锤体都不会恢复正常。分步复温并不比直接复温更有益。随着39℃复温时间的延长，更多的卵母细胞直接含有分散的染色体。
The purpose of this study was to evaluate effects of cooling and rewarming on the meiotic spindle apparatus of bovine oocytes. In experiment 1, in vitro-matured bovine oocytes were either maintained at 39% or cooled abruptly to 4 degrees C or similar to 25 degrees C. Immunohistochemical and DNA staining for visualization of microtubules and chromosomes, respectively revealed an anastral, barrel shaped spindle in bovine oocytes. Exposure to 4 degrees C for 10-20 min caused complete disappearance of the spindle. Some chromosome dispersion occurred after GO min at 4 degrees C. After exposure to similar to 25 degrees C for 30 min, 90% of oocytes appeared abnormal, having either an abnormal spindle or no spindle. In experiment 2, oocytes cooled to either similar to 25 degrees C or 4 degrees C for 30 min were rewarmed directly or in steps for 15 or 60 min. Spindles did not return to normal in most oocytes regardless of cooling temperature or rewarming scheme. Step-wise rewarming was no more beneficial than direct rewarming. More of the oocytes rewarmed directly contained dispersed chromosomes as time at 39 degrees C increased.