【摘要】：第一部分小鼠胚胎心臟房室管發(fā)育及Lef1的表達(dá)規(guī)律目的:探討小鼠胚胎心臟房室管分隔、重塑過(guò)程以及Lef1在房室管發(fā)育過(guò)程中的表達(dá)規(guī)律。方法:25只胚齡10~15d小鼠胚胎心臟連續(xù)石蠟切片,用抗心肌肌球蛋白輕鏈Ⅱa單克隆抗體(MLC2a)、抗心肌肌球蛋白輕鏈Ⅱ多克隆抗體(MLC-2)、抗Tbx3多克隆抗體(Tbx3)、抗淋巴增強(qiáng)因子1單克隆抗體(Lef1)進(jìn)行免疫組織化學(xué)染色和免疫熒光化學(xué)染色。結(jié)果:胚齡10~15d,房室管心肌呈MLC2a陽(yáng)性、MLC-2陰性,同時(shí)表達(dá)Tbx3、Lef1。胚齡10d,心內(nèi)膜墊形成。胚齡11~12d,房室管心內(nèi)膜墊內(nèi)細(xì)胞Lef1強(qiáng)表達(dá),心外膜形成。胚齡12~13d,背腹側(cè)房室管心內(nèi)膜墊相互靠近且融合形成房室瓣,心外膜來(lái)源間充質(zhì)細(xì)胞數(shù)量增加,部分表達(dá)Lef1。胚齡13d開(kāi)始,部分心外膜來(lái)源間充質(zhì)細(xì)胞穿過(guò)心肌延伸入壁側(cè)房室瓣。胚齡15d,房室瓣膜基部直接與MLC2a陽(yáng)性的房室管心肌相連。結(jié)論:小鼠胚胎房室管心肌發(fā)育為成體心臟房室環(huán)瓣膜基部的心肌;在小鼠胚胎心內(nèi)膜墊融合、房室瓣形成過(guò)程中,Lef1在心肌和心外膜來(lái)源間充質(zhì)細(xì)胞均有表達(dá)。第二部分轉(zhuǎn)錄因子Tbx3與小鼠胚胎第二生心區(qū)發(fā)育目的:探討轉(zhuǎn)錄因子Tbx3在小鼠胚胎SHF的時(shí)空表達(dá)模式及其對(duì)SHF的調(diào)節(jié)作用。方法:30只胚齡9~15d小鼠胚胎心臟連續(xù)石蠟切片,用抗胰島素增強(qiáng)子結(jié)合蛋白1多克隆抗體(isl1)、抗心肌肌球蛋白重鏈單克隆抗體(MHC)、抗心肌肌球蛋白輕鏈Ⅱa單克隆抗體(MLC2a)、抗Tbx3多克隆抗體(Tbx3)進(jìn)行免疫組織化學(xué)染色和免疫熒光化學(xué)染色。結(jié)果:胚齡10~12d,咽腹側(cè)間充質(zhì)細(xì)胞isl1陽(yáng)性表達(dá)逐漸增多,此期Tbx3的表達(dá)在咽腹側(cè)內(nèi)胚層減弱,在鄰近腹側(cè)內(nèi)胚層的間充質(zhì)細(xì)胞逐漸開(kāi)始表達(dá)。胚齡9~11d,咽腹側(cè)間充質(zhì)isl1陽(yáng)性細(xì)胞經(jīng)動(dòng)脈囊壁、心包腔背側(cè)壁延續(xù)至流出道遠(yuǎn)端壁,Tbx3在這些部位均呈陰性。而流出道遠(yuǎn)端壁MHC呈弱陽(yáng)性表達(dá)。在靜脈端,咽腹側(cè)的isl1陽(yáng)性細(xì)胞經(jīng)心背系膜延伸至DMP內(nèi),DMP內(nèi)可見(jiàn)Tbx3散在表達(dá)。胚齡12~13d,SHF isl1呈陽(yáng)性表達(dá)的細(xì)胞在動(dòng)脈端延伸進(jìn)入分隔后的動(dòng)脈壁,該部位缺乏Tbx3表達(dá);在靜脈端,DMP逐漸開(kāi)始心肌化,同時(shí)可見(jiàn)Tbx3陽(yáng)性細(xì)胞分布。胚齡15d,DMP心肌化完成,Tbx3陽(yáng)性細(xì)胞與室間隔頂部的房室束的陽(yáng)性細(xì)胞相延續(xù)。結(jié)論:Tbx3并未對(duì)SHF前體細(xì)胞的遷移與分化進(jìn)行直接的調(diào)節(jié),但可能對(duì)其存在與增殖具有調(diào)節(jié)作用,且在p SHF的調(diào)節(jié)作用與動(dòng)脈端SHF不同。
[Abstract]:The development of cardiac atrioventricular tube and the expression of Lef1 in the first part of mouse embryos: To explore the separation of atrioventricular tube, remodeling process and the expression of Lef1 in the development of atrioventricular tube in mouse embryos. Methods: 25 embryo age 10~15d mouse embryonic hearts were sectioning with continuous paraffin section and monoclonal antibodies against myosin light chain II a (MLC2a) were used. Anti myosin light chain II polyclonal antibody (MLC-2), anti Tbx3 polyclonal antibody (Tbx3) and anti lymphatic enhancement factor 1 monoclonal antibody (Lef1) were used for immunohistochemical staining and immunofluorescence staining. Results: embryo age 10~15d, MLC2a positive in atrioventricular tube, MLC-2 negative, Tbx3, Lef1. embryo age 10d, endocardial pad formation. At 11~12d, the Lef1 expression of cells in the endocardium of atrioventricular endocardium was strongly expressed and the epicardium was formed. The embryonic age was 12~13d. The endocardial pad of the dorsal ventral atrioventricular endocardium was close to each other and formed the atrioventricular valve. The number of mesenchymal cells in the epicardial source increased and the Lef1. embryo age 13D began. Some epicardial mesenchymal cells were extended through the myocardium into the wall atrioventricular flap through the myocardium. Embryo age 15d, the atrioventricular valve base is directly connected with the MLC2a positive atrioventricular canal myocardium. Conclusion: the myocardium of the mouse atrioventricular atrioventricular canal is developed into the heart of the adult atrioventricular atrioventricular ring valve base; the Lef1 is expressed in the myocardium and the epicardial mesenchymal cells in the process of the atrioventricular valve formation. The second part of the transcriptional cause is in the process of atrioventricular valve formation. Sub Tbx3 and mouse embryonic second cardiac region development aim: To explore the spatio-temporal expression pattern of transcription factor Tbx3 in mouse embryo SHF and its regulating effect on SHF. Methods: 30 embryo age 9~15d mouse embryo heart continuous paraffin section, anti insulin enhancer binding protein 1 polyclonal anti body (Isl1), anti myocardial myosin heavy chain monoclonal anti Body (MHC), anti myosin light chain light chain II a monoclonal antibody (MLC2a), anti Tbx3 polyclonal antibody (Tbx3) for immunohistochemical staining and immunofluorescence staining. Results: embryo age 10~12d, Isl1 positive expression in pharynx ventral mesodermal cells increased gradually, the expression of Tbx3 in the pharynx ventral endoderm weakened in the adjacent ventral endoderm. Isl1 positive cells of the pharynx ventral mesenchyme were 9~11d, and the pharynx ventral mesenchyme positive cells were extended to the distal end wall of the pericardial cavity, and the Tbx3 was negative in these sites. The distal end wall MHC of the outflow tract was weak positive. At the venous end, the Isl1 positive cells of the pharynx and ventral side extended to the DMP by the dorsal mesenteric membrane, and T in DMP. BX3 was expressed. The embryonic age 12~13d, SHF Isl1 positive cells extended into the wall of the separated arteries at the end of the artery, and there was a lack of Tbx3 expression in this site; at the end of the vein, DMP gradually began to cardiomyocyte, and the distribution of Tbx3 positive cells. Embryo age 15d, DMP cardiomyocyte completion, Tbx3 positive cells and positive cells of atrioventricular bundles at the top of the ventricular septum Conclusion: Tbx3 does not directly regulate the migration and differentiation of SHF precursor cells, but may regulate its existence and proliferation, and the regulatory role of P SHF is different from that of SHF in the arterial end.
【學(xué)位授予單位】：山西醫(yī)科大學(xué)
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2017
【分類號(hào)】：R329.1

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