模拟微重力效应破坏线粒体–纺锤体–染色体的协同互作诱发小鼠卵母细胞减数分裂异常
doi: 10.11728/cjss2025.05.2025-yg03 cstr: 32142.14.cjss.2025-yg03
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杨雨馨 女, 2002年1月出生于浙江省嘉兴市, 现就读于中国科学院大学材料科学与光电技术学院/中国科学院深圳先进技术研究院生物医药与技术研究所, 硕士研究生, 研究方向为空间生命科学, 空间胚胎发育. E-mail: yx.yang3@siat.ac.cn -
雷晓华 男, 1981年5月出生于江西省乐安县, 现为中国科学院深圳先进技术研究院研究员, 中国科学院大学博/硕士生导师, 主要研究方向为空间生命科学, 空间胚胎发育与干细胞研究等. E-mail: xh.lei@siat.ac.cn
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Simulated Microgravity Effects-induced Disruption of Mitochondria-spindle-chromosome Coordination Causes Meiosis Defects in Mouse Oocytes
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摘要: 采用随机定位仪模拟微重力效应, 研究重力方向连续随机改变对小鼠卵母细胞减数分裂的影响. 利用芯片装载GV期卵母细胞, 通过随机定位仪进行小鼠卵母细胞成熟培养, 分别对GV, GVBD, Pro-MⅠ,MⅠ,MⅡ各时期的线粒体、纺锤体和染色体进行荧光成像, 结果显示, 与正常重力条件相比, 模拟微重力效应导致卵母细胞成熟率显著下降32.75% (p<0.01). 荧光成像显示, 模拟微重力效应加剧了MⅠ期卵母细胞核周线粒体环状聚集现象, 同时使得MⅡ期细胞质中线粒体无规律聚集占比显著增多至71.88%. 此外, 模拟微重力效应引发MⅠ期纺锤体多极化, 导致染色体排列紊乱, 最终导致MⅡ期纺锤体组装和染色体中板宽度异常, 分别为57.58% (p<0.05) 和15.63 μm (p<0.0001). 本研究表明, 模拟微重力效应通过干扰线粒体–纺锤体–染色体的动态协调, 导致减数分裂异常和卵母细胞质量下降, 本研究对进一步研究重力条件变化对卵母细胞减数分裂的影响机制具有重要意义.Abstract: Meiosis is essential for oocyte maturation and embryonic development, representing a critical factor in mammalian reproduction. Consequently, assessing the impact of space microgravity on this process is paramount for evaluating reproductive health during long-term space missions. This article used a Random Positioning Machine (RPM) to simulate microgravity effects, examining how random changes in orientation relative to the gravity vector affect mouse oocyte meiosis. This study aims to provide critical biological references for further investigation into the damage mechanisms of space microgravity environments on oocyte meiotic division. Germinal vesicle (GV)-stage mouse oocytes were encapsulated in Polydimethylsiloxane (PDMS) chip chambers to ensure stable culture conditions and precise positional control during RPM operation. Oocytes were cultured under RPM (RPM group) and static normal gravity (NG group) conditions. Meiotic progression was tracked and quantitatively analyzed at five key developmental stages: GV (0 h), GV breakdown (GVBD, 2 h), pro-metaphase I (Pro-MⅠ, 5 h), metaphase I (MⅠ, 8 h), and metaphase II (MⅡ, 16 h). Mitochondrial distribution, spindle morphology, and chromosome alignment were quantified through confocal laser microscopy coupled with fluorescent probes. The results showed that RPM condition reduced oocyte maturation rates by 32.75% compared to normal gravity (NG) controls (p<0.01). Mitochondrial dynamics exhibited stage-specific perturbations: perinuclear clustering at MI-stage (70.00% vs. 41.18% in NG) and disorganized aggregation patterns in 71.88% of MII-stage oocytes. In addition, spindle assembly and chromosome alignment were also disrupted: multipolar spindles during MⅠ-stage caused disordered chromosome arrangement. At MⅡ, RPM group oocytes displayed exacerbated spindle defects (57.58% abnormality rate vs. 22.32% in NG, p<0.05) and widened equatorial plates (15.63 μm vs. 7.55 μm, p<0.0001). These findings suggest that the simulated microgravity effect leads to abnormal meiosis and the decline of oocyte quality through tripartite disruption of mitochondrial-spindle-chromosomal coordination. This study significantly contributes to understanding how gravity changes affect oocyte meiosis.
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Key words:
- Oocyte /
- Microgravity /
- Mitochondria /
- Spindle /
- Chromosome
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图 1 NG组和RPM组卵母细胞GVBD、Metaphase Ⅱ时期明场图像, 白色箭头表示排出极体的成熟卵母细胞 (a). NG组和RPM组卵母细胞GVBD率 (b) 与第一极体排出率 (c) 的定量分析
Figure 1. Representative bright field images in GVBD and metaphase Ⅱ oocytes under NG and RPM conditions, where the white arrows indicate mature oocytes with extruded polar bodies (a). Quantitative analysis of oocyte GVBD rates (b) and first polar body extrusion rates (c) in NG group and RPM group, respectively
图 2 NG组和RPM组卵母细胞GVBD, Prometaphase Ⅰ, Metaphase Ⅰ, Metaphase Ⅱ时期 线粒体分布代表性图像 (a) 和分布类型 (b)
Figure 2. Representative images (a) and distribution types (b) of mitochondria distribution in GVBD, Prometaphase Ⅰ, Metaphase Ⅰ, Metaphase Ⅱ of oocytes in NG and RPM groups
图 3 NG组和RPM组卵母细胞在GVBD (a), Pro-MⅠ (b), MⅠ (c), MⅡ (d) 时期的线粒体荧光强度定量; MⅡ时期卵母细胞线粒体均匀分布、核周聚集和无规律聚集的代表性图像 (e) 及其在NG和RPM条件下的比例 (f). 红色表示线粒体
Figure 3. Quantification of fluorescence intensity of mitochondria in GVBD (a), Pro-MⅠ (b), MⅠ (c), MⅡ (d) of oocytes in NG and RPM groups. Representative images of dispersive distribution, perinuclear aggregation and random aggregation of oocytes in MⅡ-stage (e) and proportions under NG and RPM conditions (f). Red represents mitochondria
图 4 NG组和RPM组卵母细胞GV, GVBD, Metaphase Ⅰ, Metaphase Ⅱ时期纺锤体形态 (a)以及染色体排列 (d) 的代表性图像; NG与RPM条件下MⅠ期卵母细胞纺锤体异常率 (b)和染色体中板宽度 (e) 定量; NG与RPM条件下MⅡ期卵母细胞纺锤体异常率 (c)和染色体中板宽度 (f) 定量. 绿色表示微管蛋白, 蓝色表示染色体
Figure 4. Representative images of spindle morphology (a) and chromosome alignment (d) in GV, GVBD, Metaphase Ⅰ and Metaphase Ⅱ stages under NG and RPM conditions, respectively. Quantification of spindle abnormalities (b) and chromosome plate width (e) in MⅠ-stage oocytes under NG and RPM conditions. Quantification of spindle abnormalities (c) and chromosome plate width (f) in MⅡ-stage oocytes under NG and RPM conditions. Green represents tubulin, and blue represents chromosome
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