Agnello Maria, Roccheri Maria Carmela, Morici Giovanni, Rinaldi Anna Maria
Dipartimento di Scienze e Tecnologie Biologiche,Chimiche e Farmaceutiche,Università degli Studi di Palermo,Viale delle Scienze,Ed. 16,Palermo 90128,Italy.
Dipartimento di Scienze e Tecnologie Biologiche,Chimiche e Farmaceutiche,Università degli Studi di Palermo,Viale delle Scienze,Ed. 16,90128 Palermo,Italy.
Zygote. 2017 Apr;25(2):205-214. doi: 10.1017/S0967199417000065. Epub 2017 Mar 9.
Sea urchin represents an ideal model for studies on fertilization and early development, but the achievement of egg competence and mitochondrial behaviour during oogenesis remain to be enlightened. Oocytes of echinoid, such as sea urchin, unlike other echinoderms and other systems, complete meiotic maturation before fertilization. Mitochondria, the powerhouse of eukaryotic cells, contain a multi-copy of the maternally inherited genome, and are involved directly at several levels in the reproductive processes, as their functional status influences the quality of oocytes and contributes to fertilization and embryogenesis. In the present paper, we report our latest data on mitochondrial distribution, content and activity during Paracentrotus lividus oogenesis. The analyses were carried out using confocal microscopy, in vivo incubating oocytes at different maturation stages with specific probes for mitochondria and mtDNA, and by immunodetection of Hsp56, a well known mitochondrial marker. Results show a parallel rise of mitochondrial mass and activity, and, especially in the larger oocytes, close to germinal vesicle (GV) breakdown, a considerable increase in organelle activity around the GV, undoubtedly for an energetic aim. In the mature eggs, mitochondrial activity decreases, in agreement with their basal metabolism. Further and significant information was achieved by studying the mitochondrial chaperonin Hsp56 and mtDNA. Results show a high increase of both Hsp56 and mtDNA. Taken together these results demonstrate that during oogenesis a parallel rise of different mitochondrial parameters, such as mass, activity, Hsp56 and mtDNA occurs, highlighting important tools in the establishment of developmental competence.
海胆是研究受精和早期发育的理想模型,但卵子在卵母细胞发生过程中的成熟能力以及线粒体行为仍有待阐明。与其他棘皮动物和其他系统不同,海胆等海胆类动物的卵母细胞在受精前完成减数分裂成熟。线粒体是真核细胞的动力源,含有多份母系遗传的基因组,并在多个层面直接参与生殖过程,因为它们的功能状态会影响卵母细胞的质量,并对受精和胚胎发育有贡献。在本文中,我们报告了关于地中海海胆卵母细胞发生过程中线粒体分布、含量和活性的最新数据。分析是使用共聚焦显微镜进行的,将处于不同成熟阶段的卵母细胞在体内与线粒体和线粒体DNA的特异性探针一起孵育,并通过免疫检测热休克蛋白56(一种著名的线粒体标记物)来进行。结果显示线粒体质量和活性平行上升,特别是在较大卵母细胞中,接近生发泡(GV)破裂时,生发泡周围的细胞器活性显著增加,无疑是出于能量目的。在成熟卵子中,线粒体活性下降,这与其基础代谢一致。通过研究线粒体伴侣蛋白热休克蛋白56和线粒体DNA获得了进一步的重要信息。结果显示热休克蛋白56和线粒体DNA都有高度增加。综合这些结果表明,在卵母细胞发生过程中,线粒体的不同参数,如质量、活性、热休克蛋白56和线粒体DNA会平行上升,突出了在建立发育能力方面的重要工具。