Christians E, Boiani M, Garagna S, Dessy C, Redi C A, Renard J P, Zuccotti M
Faculté de Médecine Vétérinaire, Université de Liège, 20, Boulevard de Colonster, Liège, 4000, Belgium.
Dev Biol. 1999 Mar 1;207(1):76-85. doi: 10.1006/dbio.1998.9157.
Mouse oocytes can be classified according to their chromatin organization and the presence [surrounded nucleolus (SN) oocytes] or absence [nonsurrounded nucleolus (NSN) oocytes] of a ring of Hoechst-positive chromatin around the nucleolus. Following fertilization only SN oocytes are able to develop beyond the two-cell stage. These studies indicate a correlation between SN and NSN chromatin organization and the developmental competence of the female gamete, which may depend on gene expression. In the present study, we have used the HSP70.1Luc transgene (murine HSP70.1 promoter + reporter gene firefly luciferase) to analyze gene expression in oocytes isolated from ovaries of 2-day- to 13-week-old females. Luciferase was assayed on oocytes after classification as SN or NSN type. Our data show that SN oocytes always exhibit a higher level of luciferase activity, demonstrating a higher gene expression in this category. Only after meiotic resumption, metaphase II oocytes derived from NSN or SN oocytes acquire the same level of transgene expression. We suggest that the limited availability of transcripts and corresponding proteins, excluded from the cytoplasm until GVBD in NSN oocytes, could explain why these oocytes have a lower ability to sustain embryonic development beyond the two-cell stage at which major zygotic transcription occurs. With this study we have furthered our knowledge of epigenetic regulation of gene expression in oogenesis.
小鼠卵母细胞可根据其染色质组织以及核仁周围是否存在Hoechst阳性染色质环(有核仁周围核仁的卵母细胞)或不存在(无核仁周围核仁的卵母细胞)进行分类。受精后,只有有核仁周围核仁的卵母细胞能够发育到二细胞期之后。这些研究表明,有核仁周围核仁的卵母细胞和无核仁周围核仁的卵母细胞的染色质组织与雌配子的发育能力之间存在相关性,这可能取决于基因表达。在本研究中,我们使用了HSP70.1Luc转基因(小鼠HSP70.1启动子+萤火虫荧光素酶报告基因)来分析从2日龄至13周龄雌性小鼠卵巢中分离出的卵母细胞中的基因表达。在将卵母细胞分类为有核仁周围核仁型或无核仁周围核仁型后,对其进行荧光素酶检测。我们的数据表明,有核仁周围核仁的卵母细胞总是表现出较高水平的荧光素酶活性,表明该类别中基因表达较高。只有在减数分裂恢复后,源自无核仁周围核仁的卵母细胞或有核仁周围核仁的卵母细胞的中期II卵母细胞才获得相同水平的转基因表达。我们认为,在无核仁周围核仁的卵母细胞中,直到生发泡破裂前,转录本和相应蛋白质在细胞质中的可用性有限,这可以解释为什么这些卵母细胞在二细胞期之后维持胚胎发育的能力较低,而二细胞期会发生主要的合子转录。通过这项研究,我们进一步了解了卵子发生过程中基因表达的表观遗传调控。