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首页> 外文期刊>Cellular Physiology and Biochemistry >The Effects of Demecolcine, Alone or in Combination with Sucrose on Bovine Oocyte Protrusion Rate, MAPK1 Protein Level and C-Mos Gene Expression Level
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The Effects of Demecolcine, Alone or in Combination with Sucrose on Bovine Oocyte Protrusion Rate, MAPK1 Protein Level and C-Mos Gene Expression Level

机译：地美西辛单独或与蔗糖联合使用对牛卵母细胞突出率，MAPK1蛋白水平和C-Mos基因表达水平的影响

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Aims: Our study aims to clarify the effects of demecolcine, alone or in combination with sucrose on bovine oocyte protrusion rate, MAPK1 protein level and c-mos gene expression level. Methods: The effects of the demecolcine concentration, treatment duration, and synergistic effects with sucrose solution on the rate of membrane protrusions of bovine oocytes were investigated. Using real-time fluorescent quantitative PCR, western blot analysis, and immunofluorescence assays, the expression of the maternal c-mos gene, the protein level of mitogen-activated protein kinase 1 (MAPK1), and the change in the localization of spindles and nuclei during the demecolcine treatment were analyzed in bovine oocytes. Results: Treatment of bovine oocytes with both demecolcine (0.6 μg/mL) and sucrose (0.05 M) for 1 h led to the highest rate of membrane protrusions, and synergistic effects were also observed. Real-time fluorescent quantitative PCR analysis revealed that the demecolcine treatment up-regulated the expression of the maternal c-mos gene. Western blot analysis indicated that the demecolcine treatment enhanced the protein level of MAPK1 in bovine oocytes. Immunofluorescence analysis indicated that the spindles and nuclei were localized at the place of the membrane protrusions. Conclusions: The present results suggest that demecolcine might contribute to the activation of the Mos/MAPK pathway and affect spindle structure. These results provide a reference for more efficient generation of enucleated bovine oocytes.
机译：目的：我们的研究旨在阐明地美沙星单独或与蔗糖联用对牛卵母细胞突出率，MAPK1蛋白水平和c-mos基因表达水平的影响。方法：研究地美辛星浓度，处理时间和蔗糖溶液协同作用对牛卵母细胞膜突出率的影响。使用实时荧光定量PCR，western印迹分析和免疫荧光测定法，分析母体c-mos基因的表达，促分裂原活化蛋白激酶1（MAPK1）的蛋白水平以及纺锤体和核的定位变化牛卵母细胞分析了地美考辛治疗期间的作用。结果：地美高辛（0.6μg/ mL）和蔗糖（0.05 M）共同处理牛卵母细胞1 h导致最高的膜突出率，并且还观察到协同作用。实时荧光定量PCR分析表明，地美考辛治疗上调了母体c-mos基因的表达。 Western印迹分析表明，地美考辛处理增强了牛卵母细胞中MAPK1的蛋白水平。免疫荧光分析表明纺锤体和核位于膜突起的位置。结论：目前的结果表明，地美可辛可能有助于Mos / MAPK途径的激活并影响纺锤体的结构。这些结果为更有效地产生去核的牛卵母细胞提供了参考。

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