發(fā)布時(shí)間：2018-05-18 07:33

  本文選題：Luman + Jab1　； 參考：《西北農(nóng)林科技大學(xué)》2016年博士論文

【摘要】：胚胎的植入是哺乳動(dòng)物受精、胚胎卵裂、致密化、囊胚形成等過(guò)程之后動(dòng)物生殖過(guò)程中非常重要的一個(gè)過(guò)程。胚胎植入是活躍的囊胚和容受性子宮之間的相互作用,這種作用對(duì)胚胎著床成功至關(guān)重要。此外,這種相互作用受到卵巢激素的調(diào)節(jié),特別是雌激素和孕激素。研究報(bào)道,多種因素在胚胎植入過(guò)程中起著重要作用,包括激素、血管活性的基因、細(xì)胞因子、生長(zhǎng)因子、細(xì)胞粘附和發(fā)育調(diào)節(jié)因子。然而,在胚胎植入過(guò)程中涉及胚胎植入前的發(fā)育與胚胎-子宮相互作用的分子途徑仍不清楚。因此,本研究旨在探討:1)Luman/CREB3蛋白質(zhì)和m RNA在小鼠胚胎植入前胚胎的時(shí)空表達(dá),和2)小鼠懷孕子宮和胚胎內(nèi)Jab1/CSN5的表達(dá)和定位。1.Luman/CREB3在附植前小鼠胚胎中的表達(dá)定位Luman/CREB3是基本亮氨酸拉鏈c AMP反應(yīng)元件結(jié)合蛋白家族內(nèi)的一種轉(zhuǎn)錄因子。Luman/CREB3蛋白與宿主細(xì)胞相互作用因子1和單純皰疹病毒蛋白質(zhì)VP16相互作用進(jìn)而誘導(dǎo)單純皰疹病毒的轉(zhuǎn)錄。目前,Luman/CREB3蛋白在生殖過(guò)程的生理功能尚不清楚。在此項(xiàng)的研究中,我們利用定量實(shí)時(shí)PCR和免疫熒光方法檢測(cè)了Luman/CREB3在小鼠卵母細(xì)胞和早期胚胎發(fā)育過(guò)程中的表達(dá)與定位。研究結(jié)果證實(shí),Luman/CREB3蛋白主要在卵泡和有絲分裂中期II階段的卵母細(xì)胞內(nèi)表達(dá),并且主要分布在卵母細(xì)胞的細(xì)胞質(zhì),細(xì)胞核和極體上。然而,Luman/CREB3蛋白質(zhì)和m RNA表達(dá)水平在合子基因組激活前有顯著提高(P0.05),并在4細(xì)胞胚胎內(nèi)表達(dá)水平最高。從8細(xì)胞胚胎開(kāi)始直到囊胚階段,Luman/CREB3蛋白質(zhì)和m RNA表達(dá)水平逐漸下降(P0.05)。Luman/CREB3蛋白同樣分布在早期胚胎的細(xì)胞核和細(xì)胞質(zhì)內(nèi)并且從2細(xì)胞胚胎到8細(xì)胞胚胎的細(xì)胞核內(nèi)高度表達(dá)。Luman/CREB3在小鼠卵母細(xì)胞和早期不同發(fā)育階段胚胎內(nèi)表達(dá)與定位的差異表明Luman/CREB3在卵母細(xì)胞成熟和早期胚胎發(fā)育過(guò)程中起著重要作用。2.Jab1/CSN5在小鼠胚胎和孕期小鼠子宮中的表達(dá)定位Jun activation domain-binding protein-1(JAB1)又叫CSN5,它與COP9信號(hào)轉(zhuǎn)導(dǎo)體的5蛋白亞基密切相關(guān),是動(dòng)植物體中保守存在的多蛋白復(fù)合體。目前,Jab1/CSN5在小鼠生殖系統(tǒng)發(fā)育過(guò)程中的作用仍不清楚。在目前的研究中,我們利用q RT-PCR,免疫組織化學(xué)和免疫熒光的方法檢測(cè)了Jab1/CSN5在胚胎和妊娠小鼠子宮內(nèi)的表達(dá)與定位。研究結(jié)果表明,Jab1/CSN5蛋白主要在子宮基質(zhì)細(xì)胞內(nèi)表達(dá),然而,在子宮腺體,子宮腔上皮細(xì)胞和蛻膜細(xì)胞內(nèi)沒(méi)有顯著表達(dá)。在懷孕5到8天的附植位點(diǎn)內(nèi)Jab1/CSN5 m RNA的表達(dá)顯著高于植入位點(diǎn)(P0.05)。從第1天開(kāi)始,Jab1/CSN5 m RNA整體的表達(dá)逐漸增高并且在第5天到達(dá)到最高。之后,從懷孕6到8天開(kāi)始逐漸下降。在小鼠胚胎和小鼠胚胎植入前和植入后(5-12天)都沒(méi)有檢測(cè)到Jab1/CSN5蛋白的表達(dá)。在懷孕6到12天卵黃囊內(nèi)臟內(nèi)胚層內(nèi)檢測(cè)到了Jab1/CSN5蛋白的明顯表達(dá)�？傊�,我們的結(jié)果證實(shí)Jab1/CSN5在基質(zhì)細(xì)胞的增殖和卵黃囊的形成過(guò)程中發(fā)揮了關(guān)鍵作用。
[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.
【學(xué)位授予單位】：西北農(nóng)林科技大學(xué)
【學(xué)位級(jí)別】：博士
【學(xué)位授予年份】：2016
【分類(lèi)號(hào)】：S814

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