Motta C M, Tammaro S, Cicale A, Indolfi P, Iodice C, Spagnuolo M S, Filosa S
Department of Evolutive and Comparative Biology, University of Naples Federico II, via Mezzocannone 8, 80134 Naples, Italy.
Mol Reprod Dev. 2001 Aug;59(4):422-30. doi: 10.1002/mrd.1049.
The present work was carried out to clarify the nature and origin of the yolk DNA present in vitellogenic oocytes of the lizard Podarcis sicula. Morphological and biochemical evidences indicate that it has an intrafollicular origin, from the apoptotic bodies resulting from follicle cells regression at the end of previtellogenesis. This conclusion is reinforced by the observation that the oocyte membrane, in in vitro experiments, is unpermeable to exogenous DNA. Biochemical evidences reveal that the yolk DNA has a low (200bp) molecular weight and this suggests that it is produced by the endonucleases typically involved in apoptotic DNA laddering. Indeed, immunocytochemical analyses demonstrate that follicle cells contain significant amounts of DNAse I. In immunoblots, carried out during different periods of the ovarian cycle, the enzyme shows a MW of about 33, 66 or 100 kDa thus indicating that its activity in the follicle of Podarcis is modulated by dimerization and/or binding to regulatory factors. Mol. Reprod. Dev. 59: 422-430, 2001.
开展本研究是为了阐明意大利壁蜥(Podarcis sicula)卵黄生成期卵母细胞中卵黄DNA的性质和来源。形态学和生化证据表明,它起源于卵泡内,来自于卵黄发生前期末期卵泡细胞退化产生的凋亡小体。体外实验中卵母细胞膜对外源DNA不可渗透这一观察结果进一步支持了这一结论。生化证据显示,卵黄DNA的分子量较低(200bp),这表明它是由通常参与凋亡DNA梯状条带形成的核酸内切酶产生的。实际上,免疫细胞化学分析表明卵泡细胞含有大量的DNA酶I。在卵巢周期不同阶段进行的免疫印迹实验中,该酶的分子量约为33、66或100 kDa,这表明其在意大利壁蜥卵泡中的活性受二聚化和/或与调节因子结合的调控。《分子生殖与发育》,第59卷,第422 - 430页,2001年。