利用ATAC-seq综合分析非包围核仁和包围核仁卵母细胞染色质可及性

IF 2.7 3区 生物学 Q3 BIOCHEMISTRY & MOLECULAR BIOLOGY Molecular Reproduction and Development Pub Date : 2023-01-04 DOI:10.1002/mrd.23668
Xiaofan Sun, Dayu Wang, Weijian Li, Qian Gao, Jingli Tao, Honglin Liu
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引用次数: 1

摘要

小鼠生发囊泡(GV)卵母细胞根据染色质形态分为包围核仁(SN)和非包围核仁(NSN)卵母细胞。NSN卵母细胞在积累足够的母体转录本后会自发转化为SN卵母细胞。SN卵母细胞表现出转录沉默。当卵母细胞成熟异常或发生在体外时,NSN卵母细胞在进入MII之前不经过SN期。小鼠卵母细胞发育迟缓,受精率低,在双细胞胚胎期发育停滞。本研究检测了GV卵母细胞的染色质结合核糖核酸聚合酶II (RNAP II)活性、新合成的RNA和染色质可及性。在SN卵母细胞中,RNAP II不与DNA结合,细胞核内不产生新rna, RNAP II的磷酸化状态不影响染色质结合活性。SN卵母细胞的可及基因数量明显低于NSN卵母细胞。不同功能基因的可及性在两种卵母细胞之间也存在差异。因此,低染色质可及性导致SN卵母细胞的转录沉默。
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Comprehensive analysis of nonsurrounded nucleolus and surrounded nucleolus oocytes on chromatin accessibility using ATAC-seq

Mouse germinal vesicle (GV) oocytes are divided into surrounded nucleolus (SN) and nonsurrounded nucleolus (NSN) oocytes based on chromatin morphology. NSN oocytes spontaneously transform into SN oocytes after accumulating enough maternal transcripts. SN oocytes show transcriptional silencing. When oocyte maturation is abnormal or takes place in vitro, NSN oocytes do not go through SN stage before proceeding to MII. Nontransitive oocytes show developmental retardation, a low fertilization rate, and arrest at the two-cell embryo stage in mice. Here, chromatin-binding ribonucleic acid polymerase II (RNAP II) activity, newly synthesized RNA, and chromatin accessibility in GV oocytes were examined. In SN oocytes, RNAP II did not bind to DNA, neo-RNA was not generated in nuclei, and the phosphorylation state of RNAP II did not affect the chromatin-binding activity. The number of accessible genes in SN oocytes was remarkably lower than that in NSN oocytes. The accessibility of different functional genes was also different between the two types of oocytes. Thus, low chromatin accessibility leads to transcriptional silencing in SN oocytes.

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来源期刊
CiteScore
5.20
自引率
0.00%
发文量
78
审稿时长
6-12 weeks
期刊介绍: Molecular Reproduction and Development takes an integrated, systems-biology approach to understand the dynamic continuum of cellular, reproductive, and developmental processes. This journal fosters dialogue among diverse disciplines through primary research communications and educational forums, with the philosophy that fundamental findings within the life sciences result from a convergence of disciplines. Increasingly, readers of the Journal need to be informed of diverse, yet integrated, topics impinging on their areas of interest. This requires an expansion in thinking towards non-traditional, interdisciplinary experimental design and data analysis.
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