Modulation of Autophagy Influences Development and Apoptosis in Mouse Embryos Developing In Vitro
Modulation of Autophagy Influences Development and Apoptosis in Mouse Embryos Developing In Vitro
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DOI：
10.1002/mrd.21331
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发表时间：
2011-07-01
影响因子：
2.5
通讯作者：
Kim, Nam-Hyung
中科院分区：
文献类型：
--
作者：
Lee, Seung-Eun;Hwang, Kyu-Chan;Kim, Nam-Hyung
自噬是蛋白质和细胞器的大量降解，对于细胞维持、细胞活力和发育至关重要，并且通常参与哺乳动物的 II 型程序性细胞死亡。本研究探讨了自噬相关基因的表达水平以及3-甲基腺嘌呤（3-MA，自噬抑制剂）或雷帕霉素（自噬诱导剂）对小鼠胚胎体外发育和凋亡的影响。 LC3对于自噬体的形成至关重要，在小鼠胚胎中广泛表达，并且在1至4个细胞中存在高水平的转录物，但在桑椹胚和囊胚阶段逐渐减少。 3-MA处理的胚胎表现出发育速率和细胞总数显着降低，但细胞凋亡率增加。此外，Lc3、Gabarap、Atg4A 和 Atg4B 的表达以及 LC3 的合成在囊胚阶段均显着减少。虽然雷帕霉素治疗不影响发育速率，但细胞数量减少，凋亡率增加。 Lc3、Gabarap、Atg4A 和 Atg4B 的表达以及 LC3 的合成也增加。使用 3-MA 或雷帕霉素调节 Lc3 mRNA 和 LC3 蛋白水平，通过破坏囊胚阶段的线粒体形态和减少 mtDNA 拷贝数，显着增加凋亡细胞死亡。有趣的是，在胚胎经 3-MA 或雷帕霉素处理后，通过 POU5F1 (OCT3/4) 免疫染色检测到的内细胞团与对照相比显着增加。这些结果表明自噬影响发育模式和细胞凋亡，并可能在早期小鼠胚胎发生中发挥作用。摩尔。重现。开发。 78: 498-509, 2011。(C) 2011 Wiley-Liss, Inc.
Autophagyis, the bulk degradation of proteins and organelles, is essential for cellular maintenance, cell viability, and development, and is often involved in type II programmed cell death in mammals. This study investigated the expression levels of autophagy-related genes and the effect of 3-methyladenine (3-MA, an autophagy inhibitor) or rapamycin (an autophagy inducer) on the in vitro development and apoptosis of mouse embryos. LC3, which is essential for the formation of autophagosomes, was widely expressed in mouse embryos, and high levels of transcript were present from 1 to 4 cells but gradually decreased through the morula and blastocyst stages. 3-MA-treated embryos exhibited significantly reduced developmental rates and total cell numbers, but increased rates of apoptosis. Furthermore, both the expression of Lc3, Gabarap, Atg4A, and Atg4B, and the synthesis of LC3 were significantly reduced at the blastocyst stage. Although rapamycin treatment did not affect developmental rates, cell numbers decreased, and the apoptosis rate increased. Expression of Lc3, Gabarap, Atg4A, and Atg4B, and synthesis of LC3 increased as well. Modulation of Lc3 mRNA and LC3 protein levels using 3-MA or rapamycin significantly increased apoptotic cell death through the disruption of mitochondrial morphology and reduction of mtDNA copy number at the blastocyst stage. Interestingly, the inner cell mass, detected by immunostaining with POU5F1 (OCT3/4) after 3-MA or rapamycin treatment of embryos, was significantly increased compared to controls. These results suggest that autophagy influences developmental patterning and apoptosis, and may play a role in early mouse embryogenesis. Mol. Reprod. Dev. 78: 498-509, 2011. (C) 2011 Wiley-Liss, Inc.