Tokmakov Alexander A, Iguchi Sho, Iwasaki Tetsushi, Fukami Yasuo, Sato Ken-Ichi
a Faculty of Life Sciences, Kyoto Sangyo University, Kamigamo Motoyama , Kita ku , Kyoto , Japan.
b Graduate School of Science, Kobe University, Rokko dai , Nada , Kobe , Japan.
RNA Biol. 2017 Mar 4;14(3):339-346. doi: 10.1080/15476286.2016.1276695. Epub 2017 Jan 3.
Cytoplasmic mRNAs are specifically degraded in somatic cells as a part of early apoptotic response. However, no reports have been presented so far concerning mRNA fate in apoptotic gametes. In the present study, we analyzed the content of various cytoplasmic mRNAs in aging oocytes and eggs of the African clawed frog, Xenopus laevis. To circumvent large gene expression variation among the individual oocytes and eggs, single-cell monitoring of transcript levels has been implemented, using multiple cytoplasmic collections and reverse transcriptase quantitative PCR. It was found that numerous cytoplasmic mRNAs, coding for proteins classified in different functional types, are robustly degraded in apoptotic Xenopus eggs, but not in aging oocytes. mRNA degradation becomes evident in the eggs after meiotic exit at the time of cytochrome c release. A strong correlation between the length of PCR amplicon and specific transcript content was observed, suggesting endonucleolytic cleavage of mRNA. In addition, it was found that mRNA deadenylation also contributes to apoptotic mRNA degradation. Altogether, these findings indicate that the global decay of mRNA represents a hallmark of apoptosis in aging Xenopus eggs. To our knowledge, this is the first description of mRNA degradation in apoptotic gamete cells.
细胞质mRNA在体细胞中作为早期凋亡反应的一部分被特异性降解。然而,迄今为止,尚未有关于凋亡配子中mRNA命运的报道。在本研究中,我们分析了非洲爪蟾(非洲爪蟾)衰老卵母细胞和卵子中各种细胞质mRNA的含量。为了规避单个卵母细胞和卵子之间较大的基因表达差异,我们采用多次细胞质采集和逆转录定量PCR技术对转录水平进行单细胞监测。结果发现,许多编码不同功能类型蛋白质的细胞质mRNA在凋亡的非洲爪蟾卵子中被大量降解,但在衰老的卵母细胞中未被降解。在细胞色素c释放时减数分裂退出后,卵子中的mRNA降解变得明显。观察到PCR扩增子长度与特定转录本含量之间存在强相关性,提示mRNA存在内切核酸酶切割。此外,还发现mRNA去腺苷酸化也有助于凋亡mRNA的降解。总之,这些发现表明mRNA的整体降解是衰老非洲爪蟾卵子凋亡的一个标志。据我们所知,这是首次对凋亡配子细胞中mRNA降解的描述。