Bachvarova R, De Leon V, Johnson A, Kaplan G, Paynton B V
Dev Biol. 1985 Apr;108(2):325-31. doi: 10.1016/0012-1606(85)90036-3.
The total RNA content of mouse oocytes, as measured by ethidium bromide fluorescence, was found to decrease by 19% during meiotic maturation (ovulated eggs contain 19% less RNA than full-grown oocytes). Consistent with these results, prelabeled stable RNA of full-grown oocytes decreased by about 20% during in vitro maturation. Polyadenylated RNA represented 19% of total prelabeled RNA in full-grown oocytes and 10% in oocytes matured in vitro, confirming previous results on in vivo prepared material. To distinguish between deadenylation and degradation for one mRNA, the amount and state of adenylation of actin mRNA was examined using Northern blots of oocyte RNA probed with a nick-translated beta-actin cloned chicken cDNA. The results showed that the amount of actin mRNA remained similar during maturation, but its molecular weight decreased slightly. Experiments in which RNA was treated with oligo(dT) and RNase H demonstrated that the actin mRNA was deadenylated during maturation, when actin synthesis is known to decline. These results indicate that the previously defined loss of bulk RNA and changes in the state of adenylation of mRNA during the first 11/2 days of embryogenesis actually begin during the 12 hr of meiotic maturation preceding fertilization.
通过溴化乙锭荧光法测量发现,小鼠卵母细胞的总RNA含量在减数分裂成熟过程中减少了19%(排卵后的卵子所含RNA比完全成熟的卵母细胞少19%)。与这些结果一致,完全成熟卵母细胞预先标记的稳定RNA在体外成熟过程中减少了约20%。聚腺苷酸化RNA在完全成熟卵母细胞中占预先标记总RNA的19%,在体外成熟的卵母细胞中占10%,证实了先前对体内制备材料的研究结果。为了区分一种mRNA的去腺苷酸化和降解,使用用缺口平移的β-肌动蛋白克隆鸡cDNA探测的卵母细胞RNA的Northern印迹法检测肌动蛋白mRNA的腺苷酸化量和状态。结果表明,肌动蛋白mRNA的量在成熟过程中保持相似,但其分子量略有下降。用寡聚(dT)和RNase H处理RNA的实验表明,当已知肌动蛋白合成下降时,肌动蛋白mRNA在成熟过程中发生去腺苷酸化。这些结果表明,先前定义的在胚胎发育的前1.5天大量RNA的损失和mRNA腺苷酸化状态的变化实际上在受精前减数分裂成熟的12小时就开始了。