Mcph1 mRNA在小鼠早期妊娠子宫中的表达研究

发布时间：2018-11-09 10:42

【摘要】：MCPH1也称为BRIT1,编码Microcephalin蛋白。MCPH1基因突变可导致一种称作原发性小头畸形的常染色体隐性遗传疾病产生。MCPH1可以抑制端粒末端转移酶的表达,是一个潜在的肿瘤抑制基因。MCPH1在维持基因组稳定性中也具有重要的作用,MCPH1与细胞周期调控过程、DNA损伤应答、细胞凋亡过程以及肿瘤发生、发展等多种细胞活动相关。本研究利用原位杂交技术分别检测了各小鼠模型子宫中Mcph1 mRNA的表达情况,小鼠模型包括早期妊娠模型、假孕模型、人工诱导蜕膜化模型和延迟着床及激活模型,根据实验结果可以总结：在早期妊娠的小鼠子宫中,Mcph1 mRNA在妊娠第1天表达较弱,且信号仅出现在子宫腔上皮；在妊娠第2、4天的子宫腔上皮和腺上皮中均有Mcph1 mRNA表达；而妊娠第3天仅在腔上皮观察到信号的表达,第2-4天的Mcph1 mRNA信号强度相比第1天均有所增强,但2-4天这三天的信号强度几乎保持同水平；在小鼠妊娠第5天,Mcph1 mRNA信号除在腔上皮表达外,也在初级蜕膜区区域出现,信号强度较强；在妊娠第6天到妊娠第8天,信号的表达仍主要集中于初级蜕膜区,并从第7天开始逐渐扩散至蜕膜区,妊娠第8天时基本覆盖整个子宫组织。在假孕模型的小鼠子宫中,Mcph1 mRNA信号在假孕第1-4天的表达情况与妊娠第1-4天基本形同,除第1,3天仅出现在在腔上皮外,在第2,4,5天既出现在腔上皮也出现在腺上皮,且Mcph1 mRNA信号强度基本没有变化。在人工诱导蜕膜化的子宫组织中,可在蜕膜区域中检测到一定强度的信号的表达,与第8天的表达情况相似；在人工蜕膜化对照组中在子宫腔上皮和腺上皮中检测到信号。在延迟着床及激活模型的小鼠子宫中,除在激活着床的子宫腔上皮和腺上皮中可观察到信号外,在其初级蜕膜化区域中也检测到了信号的表达,表达情况与妊娠第5天较相似,而在延迟着床子宫中,仅在腔上皮和腺上皮中观察到信号。另外还使用实时定量PCR的方法检测了早期妊娠小鼠模型子宫中Mcph1 mRNA表达情况,结果显示：以妊娠第1天的表达量作为对照,在妊娠1-3天内,Mcph1 mRNA表达逐渐增强,在第3天相对表达量达到最高,而在第4天相对表达量减少,在第5天Mcph1 mRNA相对表达量升高,但相对表达量仍低于第3天,在妊娠第5-8天,相对表达量呈现逐渐降低。上述研究表明,Mcph1 mRNA的表在胚胎着床期间发生明显的时间空间变化,暗示了Mcph1在该过程中可能发挥一定作用。
[Abstract]:MCPH1 also known as BRIT1, encodes Microcephalin protein. MCPH1 mutations can lead to an autosomal recessive genetic disease called primary microcephaly. MCPH1 inhibits the expression of telomere terminal transferases. MCPH1 also plays an important role in maintaining genomic stability. MCPH1 is associated with cell cycle regulation, DNA damage response, apoptosis, tumorigenesis and development. In this study, in situ hybridization was used to detect the expression of Mcph1 mRNA in the uterus of various mouse models, including early pregnancy model, pseudo pregnancy model, artificial induced decidualization model and delayed implantation and activation model. According to the experimental results, the expression of Mcph1 mRNA was weak on the first day of pregnancy, and the signal only appeared in the uterine cavity epithelium. Mcph1 mRNA was expressed in uterine cavity epithelium and glandular epithelium on the 2nd day of pregnancy. On the third day of gestation, the signal expression was observed only in the luminal epithelium. The signal intensity of Mcph1 mRNA on day 2-4 was increased compared with that on day 1, but the signal intensity was almost the same on the 3rd day of gestation compared with that on day 1. On the 5th day of gestation, Mcph1 mRNA signal was expressed not only in the luminal epithelium, but also in the primary decidua region. From the 6th day of gestation to the 8th day of pregnancy, the signal expression was mainly concentrated in the primary decidua area, and gradually spread to the decidua area from the 7th day, and the whole uterine tissue was basically covered on the 8th day of gestation. The expression of Mcph1 mRNA signal on day 1-4 of pseudopregnancy was similar to that on day 1-4 of pregnancy. Except for day 1 and day 3, it appeared only in luminal epithelium, and on day 2, it appeared in both luminal epithelium and glandular epithelium. And the Mcph1 mRNA signal intensity has not changed. In the artificial decidualized uterine tissue, a certain intensity of signal expression could be detected in the decidua region, similar to the expression on the 8th day, and in the uterine cavity epithelium and glandular epithelium in the artificial decidualized control group. In the mouse uterus with delayed implantation and activation model, the signal was observed not only in the uterine cavity epithelium and glandular epithelium, but also in the primary decidualization region, which was similar to that on the 5th day of pregnancy. In delayed implantation, signals were observed only in luminal and glandular epithelium. In addition, real-time quantitative PCR was used to detect the expression of Mcph1 mRNA in the uterus of mouse model of early pregnancy. The results showed that the expression of Mcph1 mRNA was gradually increased during the 1-3 days of gestation, compared with that of the first day of pregnancy. The relative expression reached the highest level on the 3rd day, decreased on the 4th day, and increased on the 5th day, but the relative expression level was still lower than that on the 3rd day, and decreased gradually on the 5-8 day of gestation. These results suggest that Mcph1 mRNA's surface changes in time and space during embryo implantation, suggesting that Mcph1 may play a role in this process.
【学位授予单位】：东北林业大学
【学位级别】：硕士
【学位授予年份】：2016
【分类号】：R742

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