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1.6K Views
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10:09 min
September 13th, 2022
DOI :
10.3791/64459-v
Chapters
0:05
Introduction
0:51
Nocodazole Treatment and Live Imaging
2:41
Monitoring the Pattern of Securin-GFP Degradation During Meiotic Maturation
4:42
Recruitment of MAD2 at Kinetochores by Immunofluorescence During Meiotic Maturation
7:26
Results: Evaluating Spindle Assembly Checkpoint Integrity in Mouse Oocytes
9:33
Conclusion
Transcript
非整倍性是人类早期流产的主要遗传原因。染色体分离的大多数错误发生在卵母细胞减数分裂期间。因此,评估卵母细胞中的纺锤体组装检查点对于理解卵母细胞容易出错的原因至关重要。
在这里,我们描述了三种技术,通过使用实时成像和免疫荧光的组合在检查点检查不同的关键步骤来全面评估小鼠卵母细胞中的SAC完整性。演示该程序的将是我实验室的研究助理Cecilia Blengini博士。开始制备一毫升含有诺考达唑的培养基,并用DMSO作为对照进行逆转,如手稿中所述。
对于卵母细胞成熟和实时
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Summary
染色体分离错误是卵母细胞的常见特征。因此,研究纺锤体组装检查点为生产健康卵子所需的机制提供了重要线索。本协议描述了三种互补测定,以评估小鼠卵母细胞中的纺锤体组装检查点完整性。
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