Lo Belinda K M, Archibong-Omon Agnes, Ploutarchou Panayiota, Day Anthony J, Milner Caroline M, Williams Suzannah A
Nuffield Department of Women's and Reproductive Health, University of Oxford, Women's Centre, Level 3, John Radcliffe Hospital, Oxford OX3 9DU, UK.
School of Biological Sciences, Faculty of Biology, Medicine & Health, University of Manchester, Michael Smith Building, Oxford Road, Manchester M13 9PT, UK.
Reprod Fertil Dev. 2019 Mar;31(3):529-537. doi: 10.1071/RD18209.
Cumulus-oocyte complex (COC) expansion is essential for ovulation and fertilisation and is linked to oocyte quality. Hyaluronan (HA), the major matrix constituent, is cross-linked via inter-α-inhibitor heavy chains (HCs), pentraxin 3 (PTX3) and tumour necrosis factor-stimulated gene 6 (TSG-6). All except HCs are secreted by cumulus cells in response to oocyte-secreted factors, which signal via SMAD pathways. The double mutant (DM) mouse generates oocytes lacking complex N- and O-glycans due to oocyte-specific deletion of core 1 β1,3-galactosyltransferase (C1galt1) and N-acetylglucosaminyltransferase I (Mgat1) and has modified cumulus expansion. We compared COCs before expansion (48 h-post-pregnant mare serum gonadotrophin (PMSG)) and at late-stage expansion (9 h-post-human chorionic gonadotrophin (hCG); control n=3 mice, DM n=3 per group). Using histochemistry the levels of HA, HCs, PTX3, TSG-6 and phosphorylated-SMAD1/5/8 and -SMAD2 (12-25 COCs per group) were assessed. DM COCs did not differ from Controls in cumulus size or cell density at 9 h-post-hCG; however, HA and HC levels and phosphorylated-SMAD1/5/8 were reduced. Furthermore, no correlations were found between the levels of matrix molecules and cumulus area in DM or Control samples. These data suggest that HA and HCs can support cumulus expansion provided that they are present above minimum threshold levels. We propose that oocyte-specific ablation of C1galt1 and Mgat1 may affect bone morphogenetic protein 15 synthesis or bioactivity, thereby reducing SMAD1/5/8 phosphorylation and HA production.
卵丘-卵母细胞复合体(COC)扩张对于排卵和受精至关重要,并且与卵母细胞质量相关。透明质酸(HA)是主要的基质成分,通过α-抑制因子重链(HCs)、五聚素3(PTX3)和肿瘤坏死因子刺激基因6(TSG-6)交联。除HCs外,其他成分均由卵丘细胞响应卵母细胞分泌的因子而分泌,这些因子通过SMAD途径发出信号。双突变(DM)小鼠由于核心1β1,3-半乳糖基转移酶(C1galt1)和N-乙酰葡糖胺转移酶I(Mgat1)的卵母细胞特异性缺失,产生缺乏复杂N-聚糖和O-聚糖的卵母细胞,并具有改变的卵丘扩张。我们比较了扩张前(孕马血清促性腺激素(PMSG)注射后48小时)和晚期扩张时(人绒毛膜促性腺激素(hCG)注射后9小时)的COC;对照组n = 3只小鼠,每组DM n = 3只。使用组织化学方法评估了HA、HCs、PTX3、TSG-6以及磷酸化SMAD1/5/8和-SMAD2的水平(每组12 - 25个COC)。在hCG注射后9小时,DM COC在卵丘大小或细胞密度方面与对照组没有差异;然而,HA和HCs水平以及磷酸化SMAD1/5/8降低。此外,在DM或对照样品中,未发现基质分子水平与卵丘面积之间存在相关性。这些数据表明,只要HA和HCs存在于最低阈值水平之上,就可以支持卵丘扩张。我们提出,C1galt1和Mgat1的卵母细胞特异性缺失可能会影响骨形态发生蛋白15的合成或生物活性,从而降低SMAD1/5/8磷酸化和HA产生。