Pre-fertilization zona pellucida hardening by different cross-linkers affects IVF in pigs and cattle and improves embryo production in pigs
Pre-fertilization zona pellucida hardening by different cross-linkers affects IVF in pigs and cattle and improves embryo production in pigs
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DOI：
10.1530/rep-08-0340
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发表时间：
2009-05-01
期刊：
影响因子：
3.8
通讯作者：
Coy, Pilar
中科院分区：
文献类型：
--
作者：
Canovas, Sebastian;Romar, Raquel;Coy, Pilar
透明带（ZP）硬化（抗蛋白水解）已被经典地确定为受精后事件，有助于阻止多精受精。二-（N-琥珀酰亚胺基）-3，3 '-二硫代二丙酸酯（DSP）是一种具有渗透性的胺反应性交联剂，最近被证明可诱导受精前ZP硬化并提高猪IVF生产力。本研究的目的是研究i）DSP如何影响牛的受精前ZP硬化和IVF，ii）非渗透性胺反应性交联剂（如双（磺基琥珀酰亚胺基）辛二酸酯（BS 3））是否影响牛和猪的ZP硬化和IVF，iii）如果在任一种属中证明IVF生产率改善，DSP或BS 3是否影响体外胚胎发育。牛和猪体外成熟的卵母细胞与交联剂（0.06，0.3和0.6毫克/毫升）孵育30分钟。然后，他们进行ZP消化或WE在牛，DSP和BS 3诱导ZIP硬化和降低渗透率，虽然单精子，渗透，或男性原核形成没有受到影响。在猪中，BS 3处理诱导ZP硬化，降低渗透和雄性原核形成，并增加单精子。只有当猪卵母细胞暴露于DSP时，IVF生产率才有所提高。当猪受精卵进一步体外培养时，卵裂率和囊胚形成率增加。这些结果支持了这样的观点，即在防止牛和猪的多精受精中涉及的机制具有不同的效率，并且DSP交联剂诱导的ZP硬化可能有助于提高猪胚胎生产。生殖（2009年）137 803-812
Zona pellucida (ZP) hardening (resistance to proteolysis) has been classically identified as a post-fertilization event that contributes to the block to polyspermy. Di-(N-succinimidyl)-3,3'-dithiodipropionate (DSP), a permeable amine-reactive cross-linker, was recently shown to induce pre-fertilization ZP hardening and to improve porcine IVF productivity. The objectives of this study were to investigate i) how DSP affects pre-fertilization ZP hardening and IVF in cattle, ii) if a non-permeable amine-reactive cross-linker such as bis(sulfosuccinimidyl) suberate (BS3) affects ZP hardening and IVF in cattle and pigs, and iii) whether DSP or BS3, if improvement in IVF productivity was demonstrated in either species, affects in vitro embryo development. Bovine and porcine in vitro matured oocytes were incubated with the cross-linkers (0.06, 0.3, and 0.6 mg/ml) for 30 min. Then they were subjected to ZP digestion or WE In cattle, both DSP and BS3 induced ZIP hardening and decreased the penetration rate, although monospermy, penetration, or male pronuclear formation was not affected. In pigs, BS3 treatment induced ZP hardening, decreased penetration and male pronuclear formation, and increased monospermy. IVF productivity only improved when porcine oocytes were exposed to DSP. When porcine zygotes derived from this treatment were further cultured in vitro, the cleavage and blastocyst formation rates increased. These results support the idea that mechanisms involved in the prevention of polyspermic fertilization in cattle and pigs have different efficiencies, and ZP hardening induced by DSP cross-linker may be useful for improving porcine embryo production. Reproduction (2009) 137 803-812