外泌体样囊泡对猪体外积云扩张的影响

Yuta Matsuno, Asuka Onuma, Yoshie A Fujioka, K. Yasuhara, W. Fujii, K. Naito, K. Sugiura
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引用次数: 34

摘要

细胞分泌的囊泡,如外泌体,最近被认为是细胞通讯的介质。最近对牛的一项研究表明,外泌体样囊泡参与控制积云扩张,这是正常排卵的先决条件;然而,在其他哺乳动物物种中是否也是如此尚不清楚。因此,本研究旨在研究猪卵巢卵泡中外泌体样囊泡的存在及其对体外积云扩张的影响。通过透射电镜观察、标记蛋白检测和外泌体特异性RNA谱,证实猪卵泡液(pFF)中存在外泌体样囊泡。从pFF中分离的荧光标记的外泌体样囊泡被体外结合到积云和壁粒细胞中。外泌体样囊泡不能诱导积云扩张到与促卵泡激素(FSH)诱导的程度相当。此外,外泌体样囊泡对正常扩张过程所需转录本(HAS2、TNFAIP6和PTGS2)的表达水平没有显著影响。有趣的是,fsh诱导的HAS2和TNFAIP6 mRNA的表达,而不是PTGS2 mRNA的表达,由于外泌体样囊泡的存在而显著增加;但fsh诱导的膨胀程度不受影响。此外,猪外泌体样囊泡对小鼠卵丘-卵母细胞复合物的扩增无显著影响。总的来说,目前的结果表明pFF中存在外泌体样囊泡,但它们不能有效地诱导猪的积云扩张。
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Effects of exosome-like vesicles on cumulus expansion in pigs in vitro
Cell-secreted vesicles, such as exosomes, have recently been recognized as mediators of cell communication. A recent study in cattle showed the involvement of exosome-like vesicles in the control of cumulus expansion, a prerequisite process for normal ovulation; however, whether this is the case in other mammalian species is not known. Therefore, this study aimed to examine the presence of exosome-like vesicles in ovarian follicles and their effects on cumulus expansion in vitro in pigs. The presence of exosome-like vesicles in porcine follicular fluid (pFF) was confirmed by transmission electron microscopic observation, the detection of marker proteins, and RNA profiles specific to exosomes. Fluorescently labeled exosome-like vesicles isolated from pFF were incorporated into both cumulus and mural granulosa cells in vitro. Exosome-like vesicles were not capable of inducing cumulus expansion to a degree comparable to that induced by follicle-stimulating hormone (FSH). Moreover, exosome-like vesicles had no significant effects on the expression levels of transcripts required for the normal expansion process (HAS2, TNFAIP6, and PTGS2). Interestingly, FSH-induced expression of HAS2 and TNFAIP6 mRNA, but not of PTGS2 mRNA, was significantly increased by the presence of exosome-like vesicles; however, the degree of FSH-induced expansion was not affected. In addition, porcine exosome-like vesicles had no significant effects on the expansion of mouse cumulus-oocyte complexes. Collectively, the present results suggest that exosome-like vesicles are present in pFF, but they are not efficient in inducing cumulus expansion in pigs.
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