Seminal extracellular vesicles subsets modulate gene expression in cumulus cells of porcine in vitro matured oocytes.

Seminal extracellular vesicles subsets modulate gene expression in cumulus cells of porcine in vitro matured oocytes.
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DOI:
10.1038/s41598-022-22004-7
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发表时间:
2022-11-09
期刊:
影响因子:
4.6
通讯作者:
--
中科院分区:
综合性期刊3区
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--
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精浆(SP)是一种主要由副性腺分泌物组成的液体,含有异质的细胞外囊泡(EV)群体,参与多种生殖生理过程。研究发现精浆可以在激素分泌和免疫调节方面调节卵巢功能。本研究评估了 SP-EV 子集对体外成熟 (IVM) 过程中卵丘卵母细胞复合体 (COC) 生理学调节的潜在影响。通过尺寸排阻色谱法从猪 SP 中分离出两个 SP-EV 子集:小 EV(S-EV)和大 EV(L-EV)。接下来,在不存在(对照)或存在每个 SP-EV 子集的情况下对 COC 进行 IVM,以评估 COC(PKH67-EV 标记)对它们的摄取及其对卵母细胞和卵丘细胞 (CC)(基因表达以及孕酮和雌二醇-17β 水平)的影响。 S-EV 和 L-EV 能够结合 CC,但不能结合卵母细胞。补充 L-EV 会引起 CC 中卵母细胞成熟 (HAS2) 和类固醇生成相关基因 (CYP11A1 和 HSD3B1) 转录水平的变化 (P ≤ 0.05)。当 COC 与两个 SP-EV 亚组中的任何一个进行 IVM 时,未观察到对核卵母细胞成熟以及孕酮和雌二醇-17β 水平的影响。总之,虽然 SP-EV 子集可以在 IVM 过程中被 CC 整合,但它们不会影响卵母细胞成熟,并且只有 L-EV 能够调节 CC 功能,主要是修改类固醇生成相关基因的表达。
Seminal plasma (SP), a fluid composed mainly by secretions from accessory sex glands, contains a heterogenous population of extracellular vesicles (EVs), involved in several reproductive physiological processes. Seminal plasma has been found to modulate ovary function, in terms of hormone secretion and immune regulation. This study evaluated the potential effect of SP-EV-subsets on the modulation of cumulus-oocyte-complex (COCs) physiology during in vitro maturation (IVM). Two SP-EV-subsets, small-EVs (S-EVs) and large-EVs (L-EVs), were isolated from pig SP by size-exclusion-chromatography. Next, COCs were IVM in the absence (control) or presence of each SP-EV-subset to evaluate their uptake by COCs (PKH67-EVs labelling) and their effect on oocyte and cumulus cells (CCs) (gene expression, and progesterone and estradiol-17β levels). S-EVs and L-EVs were able to bind CCs but not oocytes. Supplementation with L-EVs induced changes (P ≤ 0.05) in the transcript levels of oocyte maturation- (HAS2) and steroidogenesis-related genes (CYP11A1 and HSD3B1) in CCs. No effect on nuclear oocyte maturation and progesterone and estradiol-17β levels was observed when COCs were IVM with any of the two SP-EV-subsets. In conclusion, while SP-EV-subsets can be integrated by CCs during IVM, they do not affect oocyte maturation and only L-EVs are able to modulate CCs function, mainly modifying the expression of steroidogenesis-related genes.
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