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通过体外培养小鼠胚胎卵巢分析减数分裂进程。

Analysis of Meiotic Progression by Ex Vivo Culture of Mouse Embryonic Ovaries.

机构信息

Magee-Womens Research Institute, Department of Obstetrics, Gynecology & Reproductive Sciences, University of Pittsburgh, Pittsburgh, PA, USA.

Department of Cellular & Molecular Biology. Centro de Investigaciones Biológicas Margarita Salas (CIB-CSIC), Madrid, Spain.

出版信息

Methods Mol Biol. 2024;2818:133-145. doi: 10.1007/978-1-0716-3906-1_8.

Abstract

Oogenesis is the central process required to produce viable oocytes in female mammals. It is initiated during embryonic development, and it involves the specification of primordial germ cells (PGCs) and progresses through the activation of the meiotic program, reaching a crucial phase in prophase I before pausing at diplotene around the time of birth. The significance of meiosis, particularly the prophase I stage, cannot be overstated, as it plays a pivotal role in ensuring the formation of healthy gametes, a prerequisite for successful reproduction. While research has explored meiosis across various organisms, understanding how environmental factors, including radiation, drugs, endocrine disruptors, reproductive age, or diet, influence this complex developmental process remains incomplete. In this chapter, we describe an ex vivo culture method to investigate meiotic prophase I and beyond and the disruption of oogenesis by external factors. Using this methodology, it is possible to evaluate the effects of individual xenobiotics by administering chemicals at specific points during oogenesis. This culture technique was optimized to study the effects of two selected endocrine disruptors (vinclozolin and MEHP), demonstrating that vinclozolin exposure delayed meiotic differentiation and MEHP exposure reduced follicle size. This approach also opens avenues for future applications, involving the exploration of established or novel pharmaceutical substances and their influence on essential events during prophase I, such as homologous recombination and chromosome segregation. These processes collectively dictate the ultimate fitness of oocytes, with potential implications for factors relevant to the reproductive age and fertility.

摘要

卵子发生是雌性哺乳动物产生有活力卵子所必需的核心过程。它始于胚胎发育时期,涉及原始生殖细胞(PGCs)的特化,并通过减数分裂程序的激活进展,在前期 I 达到关键阶段,然后在出生前后的双线期暂停。减数分裂的重要性,特别是前期 I 阶段,怎么强调都不为过,因为它在确保形成健康的配子方面起着关键作用,而配子是成功繁殖的前提。尽管研究已经探索了各种生物体的减数分裂,但对于环境因素(包括辐射、药物、内分泌干扰物、生殖年龄或饮食)如何影响这个复杂的发育过程,我们的理解仍不完整。在本章中,我们描述了一种体外培养方法,用于研究减数分裂前期 I 及以后的阶段,以及外部因素对卵子发生的破坏。使用这种方法学,可以通过在卵子发生的特定时间点给予化学物质来评估单个外源性化学物质的影响。该培养技术经过优化,可用于研究两种选定的内分泌干扰物(伏马菌素 B1 和 MEHP)的影响,结果表明伏马菌素 B1 暴露延迟了减数分裂分化,而 MEHP 暴露则减少了卵泡大小。这种方法还为未来的应用开辟了途径,包括探索已建立或新型药物物质及其对前期 I 中同源重组和染色体分离等重要事件的影响。这些过程共同决定了卵子的最终适应性,对与生殖年龄和生育力相关的因素具有潜在影响。

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