Meiosis progression

  • 文章类型: Journal Article
    细胞质核糖核蛋白(RNP)颗粒是由各种RNA和蛋白质组成的无膜结构,在转录后调控中起着重要作用。虽然已知RNP颗粒在一些生物体中调节减数分裂进入,人们对它们在植物中的作用知之甚少。在这项研究中,我们观察了水稻RNA结合蛋白减数分裂的细胞质颗粒结构,这有助于控制减数分裂的进入时间,在叶原生质体和孢子母细胞中。我们与已知的细胞质RNP因子进行了共定位分析,和结构域缺失分析,以评估它们对颗粒形成和减数分裂进展的影响。还探索了跨植物物种的MEL2结构域的保护。我们的结果表明,MEL2颗粒与加工体和应激颗粒因子共定位。由LOTUS结构域和固有无序区(IDR)调节的颗粒特性的维持对于减数分裂进程中正确的MEL2功能至关重要。MEL2-like蛋白广泛存在于植物界保守的LOTUS结构域,其次是IDR,尽管它们的结构域结构不同,表明这些区域在植物物种中的功能保守性。这项研究强调了MEL2颗粒动力学的作用及其对减数分裂转变和进展的影响。
    Cytoplasmic ribonucleoprotein (RNP) granules are membraneless structures composed of various RNAs and proteins that play important roles in post-transcriptional regulation. While RNP granules are known to regulate the meiotic entry in some organisms, little is known about their roles in plants. In this study, we observed the cytoplasmic granular structures of rice RNA-binding protein MEIOSIS ARRESTED AT LEPTOTENE2 (MEL2), which contributes to the control of meiotic entry timing, in leaf protoplasts and spore mother cells. We performed colocalization analysis with known cytoplasmic RNP factors, and domain deletion analysis to assess their impact on granule formation and meiosis progression. Conservation of MEL2 domains across plant species was also explored. Our results indicated that MEL2 granules colocalized with processing body and stress granule factors. The maintenance of granule properties modulated by LOTUS domain and the intrinsically disordered region (IDR) is essential for proper MEL2 function in meiosis progression. MEL2-like proteins widely found in plant kingdom conserved LOTUS domain followed by the IDR despite their diverse domain structures, suggesting the functional conservation of these domains among plant species. This study highlights the role of MEL2 granule dynamics and its impact on meiotic transition and progression.
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  • 文章类型: Journal Article
    纺锤迁移和组装调节不对称卵母细胞分裂,这对生育至关重要。Fbxo28,作为SCF(Skp1-Cul1-F-box)泛素E3连接酶复合物的成员,在卵母细胞中特异性表达。然而,关于Fbxo28在卵母细胞减数分裂过程中的纺锤体组装和迁移中的功能知之甚少。在目前的研究中,吗啉代寡核苷酸和外源mRNA的显微注射用于敲除和挽救实验,免疫荧光染色,westernblot,利用延时共聚焦显微镜和染色体扩散来探索Fbxo28在减数分裂成熟过程中不对称分裂中的作用。我们的数据表明,Fbxo28主要位于染色体和子粒微管组织中心(aMTOC)。Fbxo28的耗尽不会影响极体挤出,但会导致纺锤体形态和迁移缺陷,表示不对称分裂的失败。此外,缺乏Fbxo28破坏了皮质和细胞质肌动蛋白的组装,并降低了ARPC2和ARP3的表达。这些缺陷可以通过外源性Fbxo28-mycmRNA补充来挽救。总的来说,这项研究表明,Fbxo28在小鼠卵母细胞减数分裂成熟过程中影响纺锤体形态和基于肌动蛋白的纺锤体迁移。
    Spindle migration and assembly regulates asymmetric oocyte division, which is essential for fertility. Fbxo28, as a member of SCF (Skp1-Cul1-F-box) ubiquitin E3 ligases complex, is specifically expressed in oocytes. However, little is known about the functions of Fbxo28 in spindle assembly and migration during oocyte meiosis I. In present study, microinjection with morpholino oligonucleotides and exogenous mRNA for knockdown and rescue experiments, and immunofluorescence staining, western blot, timelapse confocal microscopy and chromosome spreading were utilized to explore the roles of Fbxo28 in asymmetric division during meiotic maturation. Our data suggested that Fbxo28 mainly localized at chromosomes and acentriolar microtubule-organizing centers (aMTOCs). Depletion of Fbxo28 did not affect polar body extrusion but caused defects in spindle morphology and migration, indicative of the failure of asymmetric division. Moreover, absence of Fbxo28 disrupted both cortical and cytoplasmic actin assembly and decreased the expression of ARPC2 and ARP3. These defects could be rescued by exogenous Fbxo28-myc mRNA supplement. Collectively, this study demonstrated that Fbxo28 affects spindle morphology and actin-based spindle migration during mouse oocyte meiotic maturation.
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  • 文章类型: Journal Article
    长春新碱(VCR)是一种微管去稳定化疗剂,通常用于治疗患者的癌症,会引起严重的副作用,包括神经毒性。在对女性生育率的影响方面,已在VCR暴露后的患者和小鼠模型中发现卵巢毒性。然而,VCR暴露对卵母细胞质量的影响尚未阐明。我们建立了VCR暴露的体外和体内模型。结果表明,体外VCR暴露通过诱导纺锤体组装缺陷导致卵母细胞成熟失败,激活SAC,氧化应激,线粒体功能障碍,和早期凋亡,通过使用体内暴露模型证实了这一点。此外,体内VCR暴露导致非整倍性,卵母细胞-精子结合能力降低,和小鼠卵母细胞皮质中皮质颗粒的数量。一起来看,这项研究表明,VCR可导致减数分裂停滞和小鼠卵母细胞质量差。
    Vincristine (VCR) is a microtubule-destabilizing chemotherapeutic agent commonly administered for the treatment of cancers in patients, which can induce severe side effects including neurotoxicity. In context of the effects on female fertility, ovarian toxicity has been found in patients and mice model after VCR exposure. However, the influence of VCR exposure on oocyte quality has not been elucidated. We established VCR exposure in vitro and in vivo model. The results indicated in vitro VCR exposure contributed to failure of oocyte maturation through inducing defects in spindle assembly, activation of SAC, oxidative stress, mitochondrial dysfunction, and early apoptosis, which were confirmed by using in vivo exposure model. Moreover, in vivo VCR exposure caused aneuploidy, reduced oocyte-sperm binding ability, and the number of cortical granules in mouse oocyte cortex. Taken together, this study demonstrated that VCR could cause meiotic arrest and poor quality of mouse oocyte.
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  • 文章类型: Journal Article
    有机紫外线(UV)过滤器之一,八氯丙烯(OCL),主要用于各种化妆品,在土壤中经常被检测到,沉积物,水生系统和食物链。有证据证实了OCL在日本medaka中的生殖毒性。然而,关于OCL暴露对卵母细胞质量的影响知之甚少。这里,我们通过将卵母细胞暴露于体外浓度为8、22、30、40和50nM的OCL,研究了OCL对小鼠卵母细胞成熟和质量的影响。结果表明,OCL显着降低了50nM的小鼠卵母细胞胚泡破裂(GVBD)和40和50nM的极体挤压(PBE)速率。OCL暴露进一步破坏纺锤体组装和染色体排列,最后诱导非整倍体。OCL暴露也损害了线粒体功能,导致卵母细胞中ROS过量产生和凋亡。此外,OCL治疗损害了皮质颗粒的分布和卵母细胞的精子结合能力。总之,这些数据表明OCL可以干扰卵母细胞减数分裂成熟并降低卵母细胞质量。
    One of organic ultraviolet (UV) filters, Octocrylene (OCL), is mainly used in various cosmetic products, which is being frequently detected in soil, sediment, aquatic systems and food chain. There is evidence confirmed the reproductive toxicity of OCL in Japanese medaka. However, less was known about the effects of OCL exposure on oocyte quality. Here, we investigated the impacts of OCL on mouse oocyte maturation and quality by exposing oocytes to OCL in vitro at concentrations of 8, 22, 30, 40 and 50 nM. The results showed that OCL markedly reduced mouse oocyte germinal vesicle breakdown (GVBD) at 50 nM and polar body extrusion (PBE) rates at 40 and 50 nM. OCL exposure further disrupted spindle assembly and chromosome alignment, finally inducing aneuploid. Mitochondrial function was also damaged by OCL exposure, leading to ROS overproduction and apoptosis in oocytes. Moreover, OCL treatment impaired the distribution of cortical granules and sperm binding ability of oocytes. In summary, these data demonstrated that OCL could disturb the oocyte meiotic maturation and reduce oocyte quality.
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