LUMAN和JAB1在怀孕期小鼠中的表达定位

发布时间：2018-05-18 07:33

本文选题：Luman + Jab1　；参考：《西北农林科技大学》2016年博士论文

【摘要】：胚胎的植入是哺乳动物受精、胚胎卵裂、致密化、囊胚形成等过程之后动物生殖过程中非常重要的一个过程。胚胎植入是活跃的囊胚和容受性子宫之间的相互作用,这种作用对胚胎着床成功至关重要。此外,这种相互作用受到卵巢激素的调节,特别是雌激素和孕激素。研究报道,多种因素在胚胎植入过程中起着重要作用,包括激素、血管活性的基因、细胞因子、生长因子、细胞粘附和发育调节因子。然而,在胚胎植入过程中涉及胚胎植入前的发育与胚胎-子宫相互作用的分子途径仍不清楚。因此,本研究旨在探讨:1)Luman/CREB3蛋白质和m RNA在小鼠胚胎植入前胚胎的时空表达,和2)小鼠怀孕子宫和胚胎内Jab1/CSN5的表达和定位。1.Luman/CREB3在附植前小鼠胚胎中的表达定位Luman/CREB3是基本亮氨酸拉链c AMP反应元件结合蛋白家族内的一种转录因子。Luman/CREB3蛋白与宿主细胞相互作用因子1和单纯疱疹病毒蛋白质VP16相互作用进而诱导单纯疱疹病毒的转录。目前,Luman/CREB3蛋白在生殖过程的生理功能尚不清楚。在此项的研究中,我们利用定量实时PCR和免疫荧光方法检测了Luman/CREB3在小鼠卵母细胞和早期胚胎发育过程中的表达与定位。研究结果证实,Luman/CREB3蛋白主要在卵泡和有丝分裂中期II阶段的卵母细胞内表达,并且主要分布在卵母细胞的细胞质,细胞核和极体上。然而,Luman/CREB3蛋白质和m RNA表达水平在合子基因组激活前有显著提高(P0.05),并在4细胞胚胎内表达水平最高。从8细胞胚胎开始直到囊胚阶段,Luman/CREB3蛋白质和m RNA表达水平逐渐下降(P0.05)。Luman/CREB3蛋白同样分布在早期胚胎的细胞核和细胞质内并且从2细胞胚胎到8细胞胚胎的细胞核内高度表达。Luman/CREB3在小鼠卵母细胞和早期不同发育阶段胚胎内表达与定位的差异表明Luman/CREB3在卵母细胞成熟和早期胚胎发育过程中起着重要作用。2.Jab1/CSN5在小鼠胚胎和孕期小鼠子宫中的表达定位Jun activation domain-binding protein-1(JAB1)又叫CSN5,它与COP9信号转导体的5蛋白亚基密切相关,是动植物体中保守存在的多蛋白复合体。目前,Jab1/CSN5在小鼠生殖系统发育过程中的作用仍不清楚。在目前的研究中,我们利用q RT-PCR,免疫组织化学和免疫荧光的方法检测了Jab1/CSN5在胚胎和妊娠小鼠子宫内的表达与定位。研究结果表明,Jab1/CSN5蛋白主要在子宫基质细胞内表达,然而,在子宫腺体,子宫腔上皮细胞和蜕膜细胞内没有显著表达。在怀孕5到8天的附植位点内Jab1/CSN5 m RNA的表达显著高于植入位点(P0.05)。从第1天开始,Jab1/CSN5 m RNA整体的表达逐渐增高并且在第5天到达到最高。之后,从怀孕6到8天开始逐渐下降。在小鼠胚胎和小鼠胚胎植入前和植入后(5-12天)都没有检测到Jab1/CSN5蛋白的表达。在怀孕6到12天卵黄囊内脏内胚层内检测到了Jab1/CSN5蛋白的明显表达。总之,我们的结果证实Jab1/CSN5在基质细胞的增殖和卵黄囊的形成过程中发挥了关键作用。
[Abstract]:Embryo implantation is a very important process in animal reproduction after mammalian fertilization, embryo cleavage, densification, and blastocyst formation. Embryo implantation is an interaction between the active blastocyst and the receptive uterus. This effect is crucial to the success of embryo implantation. In addition, this interaction is affected by ovarian hormones. Regulation, especially estrogen and progestin. Studies have reported that many factors play an important role in embryo implantation, including hormones, vasoactive genes, cytokines, growth factors, cell adhesion and developmental regulators. However, during embryo implantation, the development of the embryo and the interaction of embryo and uterus before implantation is involved. The approach is still not clear. Therefore, the aim of this study is to explore: 1) the expression of Luman/CREB3 protein and m RNA in the preimplantation embryos of mouse embryos, and 2) the expression and localization of Jab1/CSN5 in the pregnant womb and embryo of mice. The expression of.1.Luman/CREB3 in the mouse embryos in pre implantation embryos is the basic leucine zipper C AMP reaction element. A transcription factor.Luman/CREB3 protein in the protein family interacts with host cell interaction factor 1 and herpes simplex virus protein VP16 to induce herpes simplex virus transcription. At present, the physiological function of Luman/CREB3 protein in the reproductive process is not clear. In this study, we use quantitative real-time PCR and exemptions. The expression and localization of Luman/CREB3 in the development of mouse oocytes and early embryos was detected by the immunofluorescence method. The results showed that the Luman/CREB3 protein was mainly expressed in oocytes of the follicles and in the II stage of mitosis, and mainly distributed in the cells of oocyte, the nucleus and the polar body. However, the Luman/CREB3 eggs were found. The expression level of white matter and m RNA was significantly increased before the activation of the zygote genome (P0.05), and the highest expression level in the 4 cell embryos. From the beginning of the 8 cell embryo to the blastocyst stage, the expression level of Luman/CREB3 protein and m RNA decreased gradually (P0.05).Luman/CREB3 protein was also distributed in the nucleus and cytoplasm of the early embryos and from 2 The difference between the expression and localization of.Luman/CREB3 in the mouse oocyte and the early different developmental stages indicates that the Luman/CREB3 plays an important role in the maturation of oocyte and the development of the early embryo, and the expression of.2.Jab1/CSN5 in the mouse embryo and the uterus of the mice in the pregnant mice is determined by the high expression of the cell embryo to the nucleus of the embryo in the mouse. Jun activation domain-binding protein-1 (JAB1), also called CSN5, is closely related to the 5 protein subunit of the COP9 signal transfer conductor. It is a conserved multi protein complex in the animal and plant body. At present, the role of Jab1/CSN5 in the development of the reproductive system of mice is still unclear. In the previous study, we used Q RT-PCR, immunohistochemistry. The expression and localization of Jab1/CSN5 in the uterus of embryos and pregnant mice was detected by the methods of immunofluorescence and immunofluorescence. The results showed that the Jab1/CSN5 protein was mainly expressed in the uterine stromal cells. However, there was no significant expression in the uterine glands, the uterine cavity epithelial cells and the decidua cells. The Jab1/CSN5 m RNA was within the 5 to 8 day of pregnancy. The expression was significantly higher than the implantation site (P0.05). From first days, the expression of Jab1/CSN5 m RNA increased gradually and reached the highest at fifth days. Then, it began to decline gradually from 6 to 8 days of pregnancy. The expression of Jab1/CSN5 protein was not detected before and after implantation of mouse embryos and mouse embryos (5-12 days). The eggs were pregnant from 6 to 12 days. The obvious expression of Jab1/CSN5 protein was detected in the visceral endoderm of the cysts. In conclusion, our results confirm that Jab1/CSN5 plays a key role in the proliferation of stromal cells and the formation of the yolk sac.
【学位授予单位】：西北农林科技大学
【学位级别】：博士
【学位授予年份】：2016
【分类号】：S814

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