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首页> 美国卫生研究院文献>Biology of Reproduction >Reduction of Mouse Egg Surface Integrin Alpha9 Subunit (ITGA9) Reduces the Eggs Ability to Support Sperm-Egg Binding and Fusion
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Reduction of Mouse Egg Surface Integrin Alpha9 Subunit (ITGA9) Reduces the Eggs Ability to Support Sperm-Egg Binding and Fusion

机译：减少小鼠卵表面整合素Alpha9亚基（ITGA9）降低了卵支持精卵结合和融合的能力

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The involvement of egg integrins in mammalian sperm-egg interactions has been controversial, with data from integrin inhibitor studies contrasting with evidence from knockouts showing that specific integrin subunits are not essential for fertility. An alpha4/alpha9 (ITGA4/ITGA9) integrin subfamily member has been implicated in fertilization but not extensively examined, so we tested the following three hypotheses: 1) an ITGA4/ITGA9 integrin participates in sperm-egg interactions, 2) short-term acute knockdown by RNA interference of integrin subunits would result in a fertilization phenotype differing from that of chronic depletion via knockout, and 3) detection of a fertilization phenotype is sensitive to in vitro fertilization (IVF) assay conditions. We show that mouse and human eggs express the alpha9 integrin subunit (ITGA9). RNA interference-mediated knockdown resulted in reduced levels of Itga9 mRNA and surface protein in mouse eggs. RNA interference attempts to knockdown ITGA9′s likely beta partner, beta1 (ITGB1), resulted in reduced Itgb1 mRNA but no reduction in ITGB1 surface protein. Therefore, studies using a function-blocking anti-ITGB1 antibody tested the hypothesis that ITGB1 participates in gamete interactions. Analyses of sperm-egg interactions with Itga9-knockdown eggs and anti-ITGB1 antibody-treated eggs in IVF assays using specific sperm:egg ratios revealed the following: 1) a reduction, but not complete loss, of sperm-egg binding and fusion was observed and 2) the reduction of sperm-egg binding and fusion was not detected in inseminations with high sperm:egg ratios. These data demonstrate that ITGA9 and ITGB1 participate in sperm-egg interactions but clearly are not the only molecules involved. This also shows that careful design of IVF parameters allows detection of deficiencies in gamete interactions.
机译：鸡蛋整联蛋白参与哺乳动物精子-蛋相互作用一直存在争议，整联蛋白抑制剂研究的数据与基因敲除的证据表明，特定的整联蛋白亚基对于生育不是必不可少的。 alpha4 / alpha9（ITGA4 / ITGA9）整合素亚家族成员已参与受精，但并未进行广泛检查，因此我们测试了以下三个假设：1）ITGA4 / ITGA9整合素参与精卵相互作用，2）短期急性RNA干扰整联蛋白亚基的敲除会导致受精表型不同于通过敲除引起的慢性耗竭，并且3）受精表型的检测对体外受精（IVF）分析条件敏感。我们显示，小鼠和人类的卵​​表达alpha9整合素亚基（ITGA9）。 RNA干扰介导的敲低导致小鼠卵中Itga9 mRNA和表面蛋白的水平降低。 RNA干扰试图敲低ITGA9可能的beta伴侣beta1（ITGB1），导致Itgb1 mRNA减少，但ITGB1表面蛋白没有减少。因此，使用功能阻断型抗ITGB1抗体进行的研究验证了ITGB1参与配子相互作用的假设。使用特定的精子/卵子比率在IVF分析中分析与Itga9敲除卵和抗ITGB1抗体处理的卵子的精子-卵相互作用，发现以下几点：1）精子与卵子的结合和融合减少但不完全丧失观察到和2）在高精子：卵子比率的授精中未检测到精子-卵子结合和融合的减少。这些数据表明，ITGA9和ITGB1参与了精卵相互作用，但显然不是唯一的分子。这也表明，精心设计IVF参数可以检测配子相互作用中的缺陷。

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