The secretions of oviduct epithelial cells increase the equine in vitro fertilization rate: are osteopontin, atrial natriuretic peptide A and oviductin involved?
The secretions of oviduct epithelial cells increase the equine in vitro fertilization rate: are osteopontin, atrial natriuretic peptide A and oviductin involved?
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DOI：
10.1186/1477-7827-7-129
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发表时间：
2009-11-19
期刊：
影响因子：
--
通讯作者：
Goudet G
中科院分区：
文献类型：
--
作者：
Mugnier S;Kervella M;Douet C;Canepa S;Pascal G;Deleuze S;Duchamp G;Monget P;Goudet G
输卵管上皮细胞（OEC）共培养促进人类、牛和猪物种的体外受精（IVF），但没有马物种的数据。然而，尽管有许多尝试，马IVF率仍然很低。我们的第一个目标是验证OEC对马IVF的有益影响。在哺乳动物中，输卵管蛋白已被证明与配子相互作用并在受精中发挥作用。因此，我们的第二个目标是确定参与马受精的蛋白质。在第一个实验中，我们共孵育新鲜的马精子处理的钙离子载体和体外成熟的马卵母细胞有或没有猪OEC。我们发现OEC的存在增加了IVF率。在随后的实验中，我们将马配子与OEC共孵育，我们发现1）与马和猪OEC共孵育的配子，2）完整卵丘-卵母细胞复合体与裸露卵母细胞，3）先前用人绒毛膜促性腺激素、促黄体生成素和/或雌二醇刺激的OEC与未刺激的OEC，4）体内与体外成熟卵母细胞之间的IVF率没有显著差异。为了鉴定负责OEC的积极作用的蛋白质，我们首先在马基因组中搜索编码输卵管蛋白、骨桥蛋白和心房钠尿肽A（ANP A）的基因的存在。我们发现编码骨桥蛋白和ANP A的基因存在。但输卵管素基因在马基因组进化过程中要么成为假基因，要么在马基因组序列中没有得到很好的注释。然后，我们表明，骨桥蛋白和心钠素A蛋白存在于马输卵管使用表面等离子体共振生物传感器，我们分析了它们的表达在发情周期通过Western印迹。最后，我们将马配子与纯化的骨桥蛋白或合成的ANP A共孵育。骨桥蛋白或心钠素A对IVF率无明显影响，但骨桥蛋白可轻微增加IVF率。我们的研究显示了同源和异源输卵管细胞对马IVF率的有益影响，尽管率仍然很低。需要进一步的研究来鉴定所涉及的蛋白质。我们表明，表面等离子体共振技术是有效的和强大的分析受精过程中的分子相互作用。
Oviduct epithelial cells (OEC) co-culture promotes in vitro fertilization (IVF) in human, bovine and porcine species, but no data are available from equine species. Yet, despite numerous attempts, equine IVF rates remain low. Our first aim was to verify a beneficial effect of the OEC on equine IVF. In mammals, oviductal proteins have been shown to interact with gametes and play a role in fertilization. Thus, our second aim was to identify the proteins involved in fertilization in the horse. In the first experiment, we co-incubated fresh equine spermatozoa treated with calcium ionophore and in vitro matured equine oocytes with or without porcine OEC. We showed that the presence of OEC increases the IVF rates. In the subsequent experiments, we co-incubated equine gametes with OEC and we showed that the IVF rates were not significantly different between 1) gametes co-incubated with equine vs porcine OEC, 2) intact cumulus-oocyte complexes vs denuded oocytes, 3) OEC previously stimulated with human Chorionic Gonadotropin, Luteinizing Hormone and/or oestradiol vs non stimulated OEC, 4) in vivo vs in vitro matured oocytes. In order to identify the proteins responsible for the positive effect of OEC, we first searched for the presence of the genes encoding oviductin, osteopontin and atrial natriuretic peptide A (ANP A) in the equine genome. We showed that the genes coding for osteopontin and ANP A are present. But the one for oviductin either has become a pseudogene during evolution of horse genome or has been not well annotated in horse genome sequence. We then showed that osteopontin and ANP A proteins are present in the equine oviduct using a surface plasmon resonance biosensor, and we analyzed their expression during oestrus cycle by Western blot. Finally, we co-incubated equine gametes with or without purified osteopontin or synthesized ANP A. No significant effect of osteopontin or ANP A was observed, though osteopontin slightly increased the IVF rates. Our study shows a beneficial effect of homologous and heterologous oviduct cells on equine IVF rates, though the rates remain low. Furthers studies are necessary to identify the proteins involved. We showed that the surface plasmon resonance technique is efficient and powerful to analyze molecular interactions during fertilization.
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影响因子：
3.3
作者：
BROWN, LF;BERSE, B;SENGER, DR
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