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孙小雯,沈伟,孙晓凤.2020.饥饿小鼠卵母细胞中自噬和凋亡的电镜研究.动物学杂志,55(4):485-490.
饥饿小鼠卵母细胞中自噬和凋亡的电镜研究
Morphological Observation of Autophagy and Apoptosis in Starved Mouse Oocytes Using Transmission Electron Microscopy
投稿时间:2020-03-10  修订日期:2020-06-30
DOI:10.13859/j.cjz.202004010
中文关键词:  饥饿  卵母细胞  自噬  凋亡  小鼠
英文关键词:Starvation  Oocytes  Autophagy  Apoptosis  Mice
基金项目:山东省自然科学基金项目(No. ZR2017MC033)
作者单位E-mail
孙小雯 青岛农业大学动物科技学院生殖细胞生物学实验室 青岛 266000 15192491186@163.com 
沈伟 青岛农业大学生命科学学院山东省高校动物生殖与种质创新重点实验室 青岛 266000 wshen@qau.edu.cn 
孙晓凤 青岛农业大学生命科学学院山东省高校动物生殖与种质创新重点实验室 青岛 266000 xfsun@qau.edu.cn 
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中文摘要:
      小鼠(Mus musculus domesticus)原始卵泡形成在出生后3 d内进行得最剧烈,此时有大量卵母细胞丢失。出生后不久原始卵泡库就建立起来,新生鼠都会经历一段时间饥饿再摄入母乳营养,对出生后的子鼠饥饿处理时,出现了自噬和凋亡的动态变化,自噬和凋亡都可以影响细胞的存活,这很可能与卵母细胞的大量丢失有关。在本项研究中,将对照组子鼠正常母乳喂养,处理组子鼠与母鼠分开,完全不给予母乳。分别收取饥饿1.5 d与2 d子鼠的卵巢制作电镜切片,每组3只子鼠,每只子鼠3张电镜切片,每组共统计9张切片。在电镜下观察其形态变化。通过观察发现,饥饿1.5 d的子鼠卵巢与正常1.5 d的子鼠卵巢相比,卵母细胞中的自噬小体数量显著增加。这表明,饥饿处理1.5 d促进了卵母细胞的自噬,这可能有助于维持卵母细胞的形态及存活。饥饿处理2 d的子鼠卵巢显示出不同的结果。饥饿2 d的子鼠处于生命的临界阶段,已出现小部分个体死亡。存活子鼠卵巢的电镜形态学观察发现,与正常哺乳2 d的子鼠卵母细胞相比,饥饿2 d子鼠卵母细胞中自噬小体的数量显著减少,并出现了多数卵母细胞凋亡的现象,出现许多凋亡小体。本实验研究结果显示,饥饿处理影响了原始卵泡形成过程中自噬和凋亡动态变化的过程。
英文摘要:
      The formation of primordial follicles in mice is most intense within 3 days after birth, and a large number of oocytes are lost during this period. The primordial follicle pool is established shortly after birth. Newborns will experience starvation for a period of time before ingesting breast milk nutrition. When mice are starved after birth, dynamic changes in autophagy and apoptosis occur. Both autophagy and apoptosis can affect the survival of cells, which is probably related to the loss of oocytes during the formation of primordial follicles. In this study, transmission electron microscopy (TEM) was used to research morphological alterations of ovaries starved for 1.5 d and 2 d, respectively. The ovaries of the newborn mice (Mus musculus domesticus) were collected from mice with different treatments for making electron microscope sections. Electron micrographs of ovarian sections were taken by TEM. Compared with normal control oocytes (Fig. 1a), most of the oocytes starved for 1.5 d (Fig. 1b) maintained normal cell morphology, while a large number of apoptotic or pre-apoptotic cells appeared in the ovaries of mice starved for 2 days (Fig. 1c), and almost no normal cells were observed. It was found that the number of autophagosomes in oocytes was significantly increased in mice starved for 1.5 days compared with the control (Fig. 2). All data were analyzed with GraphPad Prism software and were represent as the Mean ± SD of at least three independent experiments. Student's t-test was used to compare the difference between treatment group and the control. The ovaries of mice starved for 2 days showed evident damages to the ovary. Mice starved for 2 days were at the threshold of life, and a small number of individuals died. TEM morphological observation showed that compared with the control group, the number of autophagosomes in the oocytes of mice starved for 2 days was significantly reduced, and there were more signs of apoptosis in most oocytes (Fig. 3), which suggested that excessive starvation would cause reduced autophagy and apoptosis of a large number of oocytes. The results of this experiment show that starvation treatment affects the dynamic changes of autophagy and apoptosis during the formation of primordial follicles.
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