小鼠胚胎心脏房室管发育及Lef1的表达规律
发布时间：2018-07-29 09:26
【摘要】：第一部分小鼠胚胎心脏房室管发育及Lef1的表达规律目的:探讨小鼠胚胎心脏房室管分隔、重塑过程以及Lef1在房室管发育过程中的表达规律。方法:25只胚龄10~15d小鼠胚胎心脏连续石蜡切片,用抗心肌肌球蛋白轻链Ⅱa单克隆抗体(MLC2a)、抗心肌肌球蛋白轻链Ⅱ多克隆抗体(MLC-2)、抗Tbx3多克隆抗体(Tbx3)、抗淋巴增强因子1单克隆抗体(Lef1)进行免疫组织化学染色和免疫荧光化学染色。结果:胚龄10~15d,房室管心肌呈MLC2a阳性、MLC-2阴性,同时表达Tbx3、Lef1。胚龄10d,心内膜垫形成。胚龄11~12d,房室管心内膜垫内细胞Lef1强表达,心外膜形成。胚龄12~13d,背腹侧房室管心内膜垫相互靠近且融合形成房室瓣,心外膜来源间充质细胞数量增加,部分表达Lef1。胚龄13d开始,部分心外膜来源间充质细胞穿过心肌延伸入壁侧房室瓣。胚龄15d,房室瓣膜基部直接与MLC2a阳性的房室管心肌相连。结论:小鼠胚胎房室管心肌发育为成体心脏房室环瓣膜基部的心肌;在小鼠胚胎心内膜垫融合、房室瓣形成过程中,Lef1在心肌和心外膜来源间充质细胞均有表达。第二部分转录因子Tbx3与小鼠胚胎第二生心区发育目的:探讨转录因子Tbx3在小鼠胚胎SHF的时空表达模式及其对SHF的调节作用。方法:30只胚龄9~15d小鼠胚胎心脏连续石蜡切片,用抗胰岛素增强子结合蛋白1多克隆抗体(isl1)、抗心肌肌球蛋白重链单克隆抗体(MHC)、抗心肌肌球蛋白轻链Ⅱa单克隆抗体(MLC2a)、抗Tbx3多克隆抗体(Tbx3)进行免疫组织化学染色和免疫荧光化学染色。结果:胚龄10~12d,咽腹侧间充质细胞isl1阳性表达逐渐增多,此期Tbx3的表达在咽腹侧内胚层减弱,在邻近腹侧内胚层的间充质细胞逐渐开始表达。胚龄9~11d,咽腹侧间充质isl1阳性细胞经动脉囊壁、心包腔背侧壁延续至流出道远端壁,Tbx3在这些部位均呈阴性。而流出道远端壁MHC呈弱阳性表达。在静脉端,咽腹侧的isl1阳性细胞经心背系膜延伸至DMP内,DMP内可见Tbx3散在表达。胚龄12~13d,SHF isl1呈阳性表达的细胞在动脉端延伸进入分隔后的动脉壁,该部位缺乏Tbx3表达;在静脉端,DMP逐渐开始心肌化,同时可见Tbx3阳性细胞分布。胚龄15d,DMP心肌化完成,Tbx3阳性细胞与室间隔顶部的房室束的阳性细胞相延续。结论:Tbx3并未对SHF前体细胞的迁移与分化进行直接的调节,但可能对其存在与增殖具有调节作用,且在p SHF的调节作用与动脉端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.
【学位授予单位】：山西医科大学
【学位级别】：硕士
【学位授予年份】：2017
【分类号】：R329.1
[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.
【学位授予单位】：山西医科大学
【学位级别】：硕士
【学位授予年份】：2017
【分类号】：R329.1
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