Osteopontin reduces polyspermy during in vitro fertilization of porcine oocytes
Osteopontin reduces polyspermy during in vitro fertilization of porcine oocytes
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DOI：
10.1095/biolreprod.106.052589
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发表时间：
2006-11-01
影响因子：
3.6
通讯作者：
Prather, Randall S.
中科院分区：
文献类型：
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作者：
Hao, Yanhong;Mathialagan, Nagappan;Prather, Randall S.
本研究旨在探讨骨桥蛋白(SPP1)在猪体外受精(IVF)中的作用。初步目的是评价不同浓度(0.001、0.5、0.01、0.1和1mUg/ml)的大豆分离蛋白对体外受精过程中精子和卵母细胞的影响。结果表明，SPP1以剂量依赖的方式降低多精受精率(P<；0.05)。与对照组相比，SPP1还减少了卵母细胞中的精子数量和与透明带(ZP)结合的精子数量(P<；0.05)。与对照组相比，高剂量的SPP1(1微克/毫升)可减少穿透和雄性原核的形成(P<；0.05)。有趣的是，与对照组相比，中等剂量的SPP1提高了受精率(42.6%和44.6%比31.6%；P<；0.05)，比0.1mU g/mlSPP1提高了41%。0.1mU g/mlSPP-1处理的卵母细胞对ZP的消化能力明显高于对照组(P<；0.05)。与0和0.1pg/mlSPP1相比，1.0mU g/mlSPP1处理4h后，顶体反应的精子带与ZP结合的百分率增加。SPP1对精子活力、前向活力和精子存活率均无影响。为了确认多精受精的减少是SPP1特有的，在试管受精方案中包括了一种与妊娠相关的糖蛋白混合物，结果显示对多精受精没有影响。此外，免疫印迹显示，妊娠和非妊娠母猪在第0、3、5天的输卵管中均存在50 kDa的SPP1，且第0天的SPP1浓度高于第3天和第5天。本研究首次证明SPP1在猪多精受精的调节中起重要作用，它在体外受精过程中减少多精受精，提高受精效率。
This study was designed to determine the role of osteopontin (SPP1) in in vitro fertilization (IVF) in swine. The initial objective was to evaluate the effect of various concentrations of SPP1 (0, 0.001, 0.01, 0.1 and 1 mu g/ml) on spermatozoa and oocytes during IVF. The results demonstrate that SPP1 reduced the rate of polyspermy in a dose-dependent manner (P < 0.05). SPP1 also reduced both the number of sperm in oocytes as compared to the control and the number of spermatozoa bound to the zona pellucida (ZP) (P < 0.05). High doses of SPP1 (1 mu g/ml) reduced penetration and male pronucleus formation as compared to the control (P < 0.05). Interestingly, compared to the control group, medium doses of SPP1 increased fertilization efficiency (42.6% and 44.6% vs. 31.6%; P < 0.05), representing a 41 % improvement for 0.1 mu g/ml SPP1). The ZP of 0.1 mu g/ml SPP-1 treated oocytes was more difficult to digest than control oocytes (P < 0.05). The percentage of acrosome-reacted spermato zona bound to the ZP during IVF increased after 4 h of 1.0 mu g/ml SPP1 treatment compared to 0 or 0.1 pg/ml SPP1. SPP1 did not have an effect on sperm motility, progressive motility, and sperm viability. To confirm that the reduction of polyspermy was specific to SPP1, a mixture of pregnancy-associated glycoproteins was included in the IVF protocol and shown to have no effect on polyspermy. Furthermore, Western blotting demonstrated that a 50-kDa SPP1 form was present in the oviducts on Days 0, 3, and 5 in pregnant and nonpregnant gilts, and the concentration of SPP1 on Day 0 was higher than on Days 3 and 5. The current study represents the first report to demonstrate that SPP1 plays an important role in the regulation of pig polyspermic fertilization; it decreases polyspermy and increases fertilization efficiency during IVF.