神经嵴细胞和ISL-1阳性细胞在小鼠胚胎心动脉端发育过程中的分布特点

发布时间：2018-06-17 22:30

本文选题：激活蛋白-2α + 胰岛因子-1　；参考：《山西医科大学》2012年硕士论文

【摘要】：研究背景：心神经嵴细胞和第二生心区来源的胰岛因子-1（ISL-1）阳性心前体细胞共同参与了脊椎动物弓动脉和心流出道的形态发生。转录因子ISL-1是第二生心区的主要标记蛋白，促进第二生心区心前体细胞增殖、存活和迁移。第二生心区ISL-1阳性细胞是多潜能细胞，在胚胎发育的不同时空阶段，受不同转录因子及信号系统调节，可以分化为心肌细胞、平滑肌细胞以及其它类型的细胞。第一生心区来源的原始心管形成后，第二生心区的ISL-1阳性间充质细胞以及起自听板和第三体节间的心神经嵴细胞向原始心管动脉端迁移，参与弓动脉及原始心管流出道的形态发生。去除心神经嵴，或干扰第二生心区的发育，产生相同或类似的弓动脉、流出道发育畸形，包括动脉干永存、肺动脉狭窄、右心室双流出道等，但有关第二生心区ISl-1阳性细胞与心神经嵴细胞在胚胎心流出道形态发生过程中的时空分布关系研究较少。转录因子激活蛋白-2α（AP-2α）是迁移前及迁移中的神经嵴细胞的良好标记蛋白，尚未见迁移过程中的AP-2α阳性神经嵴细胞与第二生心区前体细胞时空分布关系的报道。本实验用免疫组化方法观察了不同发育时期ISL-1阳性心前体细胞和AP-2α阳性心神经嵴细胞在小鼠胚胎弓动脉、心流出道发育过程中的时空分布特点，为进一步探讨因流出道发育异常导致的先心病发病机制提供了实验依据。研究目的：探讨迁移中的神经嵴细胞、ISL-1阳性心前体细胞在小鼠胚胎心动脉端发育过程中的分布特点。研究方法：胚龄8～12d小鼠胚胎心脏连续石蜡切片，选用抗α-平滑肌肌动蛋白（α-SMA）抗体，抗心肌肌球蛋白重链（MHC）抗体，抗转录因子ISL-1抗体，抗激活蛋白AP-2α抗体，进行免疫组织化学染色。研究结果：胚龄8d，AP-2α阳性神经嵴细胞主要表达于神经褶外胚层及邻近耳板外胚层。ISL-1阳性心前体细胞分布于前肠及神经沟两侧中胚层，并沿原始心包腔背侧脏壁中胚层延伸至发育中的心管背侧壁，构成第二生心区，与MHC、α-SMA阳性心肌细胞相延续。胚龄9～10d， AP-2α阳性神经嵴细胞在前肠两侧沿弓动脉逐渐迁移至流出道头端心内膜垫，并形成主肺动脉隔雏形。胚龄11d，AP-2α阳性神经嵴细胞由前肠两侧沿第四对弓动脉向腹侧延伸，包绕前肠腹侧及外侧壁并突向动脉囊腔，，形成AP-2α阳性的主肺动脉隔。胚龄9d，ISL-1阳性细胞分布于前肠两侧及鳃弓核心间充质，向流出道添加心肌细胞。胚龄10～11d，ISL-1阳性细胞在前肠腹侧正中间充质中聚集、与AP-2α阳性细胞共同参与主肺动脉隔形成，但并未扩展至前肠侧壁与弓动脉之间的间充质。心包腔背侧脏壁中胚层及鳃弓核心间充质的ISL-1阳性细胞与流出道α-SMA或MHC阳性心肌相延续。胚龄12d，主肺动脉隔及流出道心内膜垫失去AP-2α及ISL-1阳性表达，转变为α-SMA阳性表达结构。前肠腹侧ISL-1阳性细胞与升主动脉和肺动脉干壁ISL-1强阳性细胞延续，升主动脉和肺动脉干开始表达α-SMA。研究结论: AP-2α是心神经嵴细胞的特殊标记物，AP-2α阳性心神经嵴细胞和第二生心区来源的ISL-1阳性心前体细胞共同参与了小鼠心动脉端的形态发生，是发育中的心流出道所必需的，AP-2α缺失会导致小鼠胚胎心流出道的发育异常。胚龄10～11d，ISL-1阳性心前体细胞与AP-2α阳性细胞共同参与了主肺动脉隔及心内膜垫的形成。胚龄11d后，AP-2α阳性神经嵴细胞仅存在于第六对弓动脉周围，ISL-1阳性主肺动脉隔逐渐与流出道嵴融合，并转变为α-SMA阳性；ISL-1强阳性的主肺动脉干壁开始表达α-SMA，表明主肺动脉隔和主肺动脉干壁均开始向平滑肌方向分化。
[Abstract]:Background: cardiac neural crest cells and islet factor -1 (ISL-1) positive cardiac precursor cells derived from the second cardiac region jointly participate in the morphogenesis of the arch and cardiac outflow of vertebrates. The transcription factor ISL-1 is the main marker protein in the second cardiac region, promoting the proliferation, survival and migration of the second cardiac precardiac cells. Second heart ISL-1 positive cells are pluripotent cells. They are regulated by different transcription factors and signal systems in different temporal and spatial stages of embryonic development. They can be differentiated into cardiomyocytes, smooth muscle cells and other types of cells. After the formation of the original cardiac tube from the first cardiac region, the ISL-1 positive mesenchymal cells in the second heart region and from the auditory plate are from the first cardiac region. The crest cells of the internode and the third body migrate to the end of the original canal artery, participate in the morphogenesis of the arch artery and the original cardiac canal flow, remove the crest of the heart, or interfere with the development of the second heart region, produce the same or similar arch arteries, and the deformity of the outflow tract, including the immortality of the arteries, the stenosis of the pulmonary artery, and the double flow of the right ventricle, etc. However, there are few studies on the spatial and temporal distribution of ISl-1 positive cells in the second cardiac region and the cardiac neural crest cells during the embryonic cardiac outflow formation. The transcriptional factor activating protein -2 alpha (AP-2 a) is a good marker for the neural crest cells in the migration and migration, and the AP-2 - positive neural crest cells in the migration process have not yet been seen. The temporal and spatial distribution of the precursor cells in two cardiac region was reported. In this experiment, the temporal and spatial distribution characteristics of ISL-1 positive cardiac precursor cells and AP-2 alpha positive crest cells at different developmental stages were observed in the development process of the heart flow path. The pathogenesis of the disease provides an experimental basis.
Objective: To investigate the distribution of ISL-1 positive cardiac progenitor cells in the developing process of embryonic heart vein.
Methods: the continuous paraffin section of embryo heart of 8 ~ 12D mouse embryo was studied. Anti alpha smooth muscle actin (alpha -SMA) antibody, anti myosin heavy chain (MHC) antibody, anti transcription factor ISL-1 antibody and anti activator protein AP-2 alpha antibody were used for immunohistochemical staining.
The results showed that the embryo age 8D, AP-2 alpha positive neural crest cells were mainly expressed in the nerve fold ectoderm and the adjacent ectoderm.ISL-1 positive cardiac precursor cells distributed in the mesoderm on both the foregut and the nerve trench, and extended to the dorsal wall of the developing heart tube along the dirty wall of the original pericardial cavity, forming the second heart region, with MHC and alpha -SMA positive. The phase of the cardiac myocytes was 9 ~ 10d, and the AP-2 alpha positive neural crest cells migrated to the endocardium on both sides of the foregut to the end endocardial pad of the outflow tract, and formed the embryonic form of the main pulmonary artery septum. The embryo was 11d, and the AP-2 a positive neural crest cells extended from both sides of the foregut along the fourth pairs of arch arteries to the ventral side of the foregut and the lateral wall of the foregut and to the artery inrush to the artery. AP-2 alpha positive main pulmonary artery septum was formed in the cyst cavity. The embryo age was 9D. The ISL-1 positive cells were distributed on the two sides of the foregut and the core of the branchial arch, and the cardiomyocytes were added to the outflow tract. The embryo was 10 to 11d. The ISL-1 positive cells gathered in the mesenchyme in the ventral front of the foregut, and were involved in the formation of the main pulmonary septum with the AP-2 alpha positive cells, but did not extend to the foregut. The intermesenchyme between the lateral wall and the arch artery. The ISL-1 positive cells of the mesoderm and the core of the branchial arch in the dorsum of the pericardial cavity continued with the positive myocardium of the outflow channel alpha -SMA or MHC. The embryonic age was 12D. The main pulmonary artery septum and the outflow tract endocardium mats lost the positive expression of AP-2 alpha and ISL-1, and changed to the positive expression of alpha -SMA. The ventral ISL-1 positive of the foregut was thin. ISL-1 positive cells in the ascending aorta and pulmonary artery trunk were extended, and the ascending aorta and pulmonary artery began to express alpha -SMA..
Conclusions: AP-2 alpha is a special marker for crest cells of the heart. The AP-2 alpha positive crest cells and the ISL-1 positive cardiac precursor cells derived from the second cardiac region are involved in the morphogenesis of the cardiac pulse end of the mice, which are necessary for the developing cardiac flow out of the heart. The absence of AP-2 a may lead to abnormal development of the embryonic cardiac outflow in mice. 10 ~ 11d, ISL-1 positive cardiac precursor cells and AP-2 alpha positive cells co participated in the formation of the main pulmonary artery septum and endocardial pad. After 11d, AP-2 positive neural crest cells only existed around sixth pairs of arch arteries, and the ISL-1 positive main pulmonary septum gradually fused with the outflow ridge, and transformed into alpha -SMA positive; ISL-1 that was strongly positive for the main pulmonary artery. The dry wall began to express alpha -SMA, indicating that the main pulmonary artery septum and main pulmonary artery wall began to differentiate into smooth muscle.
【学位授予单位】：山西医科大学
【学位级别】：硕士
【学位授予年份】：2012
【分类号】：R321

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