摘要:在哺乳动物的卵母细胞减数分裂过程中,纺锤体从卵母细胞中央到皮质层是建立皮质层极性的第一步,这也是连续两轮高度不对称的细胞分裂中必不可少的过程。因此纺锤体在不对称减数分裂过程中十分必要,它确保了卵子中遗传物质的稳定性并为受精做好准备。KIF4A 是新近发现的驱动蛋白家族成员之一,是染色体凝集及整列中发现的一个重要蛋白,在有丝分裂后期转运PRCI蛋白到纺锤体中央区参与微管捆绑作用。但是,KIF4在小鼠卵子减数分裂过程中的作用机制还不清楚。因此,在本实验中,为了确定KIF4a对小鼠卵子成熟过程的影响,我们采用免疫荧光技术检测到KIF4a主要在卵子细胞中的纺锤体附近表达;而激光共聚焦结果表明,KIF4a活性被抑制后将导致纺锤体和染色体异常;通过显微镜对注射KIF4a抗体的卵子细胞进行观察,发现KIF活性被抑制后将导致卵母细胞的极体无法正常排出。因此,KIF4a可能通过影响纺锤体的形成进而影响卵母细胞的减数分裂。29704
毕业论文关键词:KIF4a;纺锤体;卵母细胞成熟
Molecular regulation mechanism of KIF4 on mouse egg maturation
Abstract:During mammalian oocyte meiosis, the meiotic spindle from the central oocyte to subcortical localization is the first step for establishment of cortical polarity, which is essential for two consecutive rounds of unique asymmetric cell pisions. So the spindle is necessary in the process of asymmetric meiosis, which ensures the stability of the genetic material in the oocyte and is ready for fertilization. KIF4A is one of the newly discovered kinesin family members. It is an important protein founded in chromosome agglutination and sequestration. Transfection of PRCI protein into the central region of mitosis is involved in microtubule binding at the later stage of mitosis. However, the mechanism of KIF4 in the oocyte of mouse is not clear. Therefore, in order to determine the effect of KIF4a on the maturation process of oocyte of mouse , we used immunofluorescence to detect the expression of KIF4a in the vicinity of the spindle in the oocytes. And the laser confocal results showed that KIF4a activity was inhibited will lead to abnormalities of spindle and chromosome ; Through observe the oocytes which injected by KIF4a antibody by use microscope. We found KIF4a activity will be inhibited after the oocyte caused by the polar body cannot be properly discharged. Thus, KIF4a may affect the meiotic pision of oocytes by affecting the formation of the spindle.
Key words: KIF4a;spindle;oocyte meiosis
目 录
摘要1
关键词1
Abstract1
Key words1
引言1
1材料与方法2
1.1材料 2
1.1.1实验动物2
1.1.2抗体和试剂2
1.1.3仪器设备2
1.2方法 3
1.2.1卵母细胞的收集和体外培养3
1.2.2 显微注射KIF4a抑制剂处理卵母细胞3
1.2.3免疫荧光染色和激光共聚焦分析3
1.2.4统计分析3
2结果与分析3
2.1 KIF4a的定位与表达3
2.2 KIF4a影响小鼠卵母细胞成熟过程中的极体排出4
2.3 KIF4a影响纺锤体在卵母细胞减数分裂过程中的形成5
3讨论 6
致谢7
参考文献7
KIF4基因对小鼠卵子成熟的分子调控机制
引 言
随着现代畜牧业的的进步,使得我们对动物繁育和生殖调控技术的需求越来越高。体外受精技术越来越成熟,我们对卵母细胞发育机制的了解也越发的深入,解决了许多动物繁育甚至人类生殖障碍方面的问题。但试管动物以及试管婴儿的出生率依旧无法与自然分娩的相比,大约只有20% [1]。而试管动物出生率的决定因素之一即是卵母细胞质量的好坏和其成熟分裂的程度。因此卵母细胞成熟机制的研究对于畜牧发展和辅助生殖有着十分重要的意义。 KIF4基因对小鼠卵子成熟的分子调控机制:http://www.youerw.com/shengwu/lunwen_25045.html