Tae Jin Cheol, Kim Eun Young, Jeon Kilsoo, Lee Keum Sil, Lee Chang Hyun, Kim Yeon Ok, Park Se Pill, Kim Nam-Hyung
Mirae Biotech Co., Seoul, Korea.
Mol Reprod Dev. 2008 Aug;75(8):1300-6. doi: 10.1002/mrd.20777.
In order to understand the mechanism by which mitogen-activated protein kinase (MAPK) regulates fertilization, we examined the effect of the MAPK pathway inhibitor U0126 on polyspermy, cortical granule reaction and mitosis in bovine oocytes during and after fertilization. Oocytes were treated with 30 microM U0126 for 30 min prior to insemination, or from 15 to 27 hr following insemination. Western blotting with antibodies that detect active, phosphorylated MAPK revealed that MAPK activity was decreased in U0126 treated oocytes. Oocytes that were treated with U0126 before insemination displayed a significantly higher incidence of polyspermic penetration and incomplete cortical granule reaction than that observed in untreated oocytes (P < 0.05). Exposure of oocytes to 30microM U0126 15-27 hr after insemination induced aberrant microtubule assembly and cell division, often resulting in the formation of two or three daughter cells with altered shapes and sizes. These results suggest that an ERK-like cascade is part of a mechanism that controls cortical granule reaction and the formation of the mitotic spindle following sperm penetration in the bovine.
为了了解丝裂原活化蛋白激酶（MAPK）调节受精的机制，我们研究了MAPK通路抑制剂U0126对牛卵母细胞在受精期间及之后的多精受精、皮质颗粒反应和有丝分裂的影响。在授精前，用30微摩尔/升的U0126处理卵母细胞30分钟，或在授精后15至27小时进行处理。用检测活性磷酸化MAPK的抗体进行蛋白质免疫印迹分析表明，经U0126处理的卵母细胞中MAPK活性降低。授精前用U0126处理的卵母细胞，其多精穿透发生率和皮质颗粒反应不完全的发生率显著高于未处理的卵母细胞（P<0.05）。授精后15至27小时，将卵母细胞暴露于30微摩尔/升的U0126中会诱导异常的微管组装和细胞分裂，常常导致形成两个或三个形状和大小改变的子细胞。这些结果表明，在牛中，一种类似ERK的级联反应是控制皮质颗粒反应和精子穿透后有丝分裂纺锤体形成机制的一部分。
