Dnm1l mRNA在小鼠早期妊娠子宫中的表达研究

发布时间：2018-01-21 16:49

本文关键词： Dnmll 胚胎着床原位杂交实时荧光定量PCR　出处：《东北林业大学》2014年硕士论文　论文类型：学位论文

【摘要】：Dnmll属于动力蛋白GTP酶超家族,是线粒体分裂体系的重要组成成分。在不同物种中,Dnmll在与多种分子相互作用后,可以定位于线粒体并组装成高级结构,引起膜的收缩和分裂。Dnmll功能的消失会增强线粒体的融合和线粒体之间的连通性。近期研究发现,Dnmll在细胞凋亡过程和多种疾病中也具有重要作用。本研究应用原位杂交的方法分别检测了Dnmll mRNA在早期妊娠、假孕、人工蜕膜化、延迟着床及激活等模型的小鼠子宫中的表达情况,并利用实时荧光定量PCR的方法研究Dnmll mRNA在小鼠早期妊娠子宫中相对表达量的变化。原位杂交结果显示,Dnmll mRNA在妊娠第1-4天的子宫腔上皮和腺上皮中均有表达,第1天的信号强度较弱,第2-4天的子宫腔上皮和腺上皮中的信号强度相比第1天均有所增强；在妊娠第5天,除在腔上皮和腺上皮中表达外,在子宫基质中也呈现弱表达,且主要集中在初级蜕膜区；在假孕模型中,假孕第1天子宫的腔上皮叶1可观察到信号的出现,在假孕第2-5天,子宫腔上皮和腺上皮中均有信号表达,信号强度基本没有改变；在激活着床的子宫组织中,信号的表达情况与妊娠第5天相似,除在腔上皮和腺上皮中可观察到信号外,在初级蜕膜化区域中也检测到了少量信号的表达；在延迟着床子宫组织中,只在腔上皮和腺上皮中观察到信号。在妊娠第6-8天,信号主要集中于蜕膜区,且随着蜕膜化组织的发展,信号的范围也不断扩大,最后几乎涵盖大部分基质细胞。在人工诱导蜕膜化的子宫组织中,在蜕膜区可检测到信号的表达,且表达情况与妊娠第8天的情况相似,在对照组的子宫组织中,只在腔上皮和腺上皮中检测到信号,且信号强度较低。实时荧光定量PCR结果显示：Dnmll mRNA在妊娠第1-3天内,表达逐渐增强,在第三天是表达量最高,在妊娠第4到6天时,表达又逐渐减弱,在妊娠第6至8天时,Dnmll mRNA的表达量比较接近。综上所述,Dnmll mRNA在小鼠早期妊娠的子宫组织中发生时空特异性表达,并且相对表达量也发生变化,Dnmll mRNA在小鼠子宫基质中的表达变化受到胚胎着床的调节。
[Abstract]:Dnmll belongs to the superfamily of dynamic protein GTP, which is an important component of mitochondrial mitosis system. In different species, Dnmll interacts with a variety of molecules. It can be located in mitochondria and assembled into advanced structure, causing membrane contraction and fission. The disappearance of Dnmll function will enhance mitochondrial fusion and mitochondrial connectivity. Dnmll also plays an important role in the process of apoptosis and many diseases. In this study, in situ hybridization was used to detect the expression of Dnmll mRNA in mouse uterus of early pregnancy, pseudopregnancy, artificial decidualization, delayed implantation and activation. The changes of relative expression of Dnmll mRNA in early pregnancy uterus of mice were studied by real-time fluorescence quantitative PCR. Dnmll mRNA was expressed in uterine cavity epithelium and glandular epithelium on the 1st and 4th day of pregnancy, and the signal intensity was weak on the 1st day. The signal intensity of uterine cavity epithelium and glandular epithelium on day 2-4 was higher than that on day 1. On the 5th day of pregnancy, the expression was weak in the uterine matrix, and mainly in the primary decidua area, except in the luminal epithelium and glandular epithelium. In the model of pseudopregnancy, the signal was observed in the luminal epithelium of uterus on the first day of pseudopregnancy, and the signal expression was found in the epithelium of uterine cavity and glandular epithelium on day 2-5 of pseudopregnancy, and the signal intensity was not changed. The signal expression was similar to that in the 5th day of pregnancy in the uterine tissues activated by implantation. In addition to the signals observed in the luminal and glandular epithelium, a small number of signals were also detected in the primary decidualization region. The signal was observed only in the luminal and glandular epithelium in the delayed implantation uterine tissue. On the 6th to 8th day of pregnancy, the signal was mainly concentrated in decidua area, and the range of signal was enlarged with the development of decidualized tissue. Finally, most of the stromal cells were almost covered. In the artificial induced decidualization of uterus, signal expression could be detected in decidua, and the expression was similar to that on the 8th day of pregnancy. In the control group, the signal was detected only in the luminal and glandular epithelium, and the signal intensity was low. The results of real-time fluorescence quantitative PCR showed that the mRNA was within 1-3 days of gestation. The expression of Dnmll mRNA was higher on the third day, decreased gradually on the 4th to 6th day of gestation, and the expression of Dnmll mRNA was similar at the 6th to 8th day of gestation. To sum up, the expression of Dnmll mRNA in the uterus of early pregnancy in mice was spatio-temporal specific, and the relative expression of Dnmll mRNA was also changed. The expression of Dnmll mRNA in mouse uterine matrix was regulated by embryo implantation.
【学位授予单位】：东北林业大学
【学位级别】：硕士
【学位授予年份】：2014
【分类号】：R321

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