The effects of plasma-derived extracellular vesicles on cumulus expansion and oocyte maturation in mice
The effects of plasma-derived extracellular vesicles on cumulus expansion and oocyte maturation in mice
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DOI：
10.1016/j.repbio.2021.100593
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发表时间：
2021-12-11
影响因子：
2.1
通讯作者：
Roshangar, Leila
中科院分区：
文献类型：
--
作者：
Javadi, Maryam;Rad, Jafar Soleimani;Roshangar, Leila
卵丘细胞扩增是可受精卵母细胞排卵所必需的。细胞外囊泡（EV）是一种存在于多种体液中的双层脂质膜囊泡，在生物学过程中起着重要作用。本研究旨在研究血浆来源的EV对卵丘扩张和卵母细胞体外成熟（IVM）的影响。使用超离心法从雌性小鼠的血浆中分离EV。通过透射电子显微镜（TEM）和动态光散射（DLS）分析了EV的形态和尺寸。蛋白质印迹法使我们能够识别EV的CD 63、CD 81、CD 9和HSP 70蛋白标志物;使用实时聚合酶链反应评估与卵丘细胞扩增相关的基因的表达，包括透明质酸合酶2（Has 2）和胰十二烷内过氧化物合酶2（Ptgs 2）。用Dil染料标记的血浆衍生的EV在IVM期间成功地与卵丘细胞结合。血浆来源的EV显著诱导卵丘扩张和卵母细胞成熟。与其他组相比，EV治疗组中达到MII期的卵母细胞百分比显著更高。尽管表皮生长因子（EGF）治疗显着增加了卵丘-卵母细胞复合体（COCs）中的卵丘扩张，但其影响小于血浆来源的EV。此外，从血浆中产生的EV显著增强了卵丘-卵母细胞复合体中Has 2和Ptgs 2 mRNA的表达。这项研究表明，来自血浆的EV能够促进卵丘扩张和卵母细胞成熟。(c)2021年生殖生物学学会和波兰科学院动物生殖和食品研究所在奥尔斯廷。Elsevier B. V.出版，保留所有权利。
Cumulus cell expansion is required for the ovulation of a fertilizable oocyte. Extracellular vesicles (EVs) are bilayer-lipid membrane vesicles that may be found in a variety of bodily fluids and play an important role in biological processes. This study aimed to examine the effects of plasma-derived EVs on cumulus expansion and in vitro maturation (IVM) of the oocyte. EVswere isolated using ultracentrifugation from the plasma of female mice. The morphology and size of EVs were analyzed by transmission electron microscopy (TEM) and dynamic light scattering (DLS). Western blotting allowed us to identify CD63, CD81, CD9, and HSP70 protein markers of EVs; the expression of the genes related to cumulus cell expansion, including hyaluronan synthase 2 (Has2) and prostaglandinendoperoxide synthase 2 (Ptgs2), were assessed using real-time polymerase chain reaction. Plasma-derived EVs labeled with Dil dye were successfully incorporated with cumulus cells during IVM. Plasma-derived EVs significantly induced cumulus expansion and maturation of oocytes. The percentage of oocytes that reached the MII stage was significantly greater in the EVs treatment group compared with other groups. Although treatment with epidermal growth factor (EGF) significantly increased cumulus expansion in cumulus-oocyte complexes (COCs), the impact was less than that seen with plasma-derived EVs. Furthermore, EVs generated from plasma substantially enhanced Has2 and Ptgs2 mRNA expression in the cumulus-oocyte complex. This research indicates that EVs derived from plasma are capable of promoting cumulus expansion and oocyte maturation. (c) 2021 Society for Biology of Reproduction & the Institute of Animal Reproduction and Food Research of Polish Academy of Sciences in Olsztyn. Published by Elsevier B.V. All rights reserved.