EVC2在早期胚胎发育中的作用研究

发布时间：2018-05-19 14:50

本文选题：EVC2 + 胚胎　；参考：《广东药学院》2011年硕士论文

【摘要】：Ellis-van Creveld综合症(Ellis-van Creveld Syndrome、EvC)又称为软骨外胚层发育不良症(chondroectodermal dysplasia),是一种常染色体隐性遗传病,临床主要表现为软骨营养障碍,多指(趾),外胚层发育不良,心脏发育异常。60%的EvC患者患有先天性心脏病。EVC2(limbin,LBN)是一种整合蛋白,它在正常的生长和发育过程中起着极其重要的作用。研究表明:出生前,它在心脏、肺、肝脏、肾脏、胰腺、运动肌(骨骼肌)等多种组织器官中表达。EVC2基因或EVC基因任何一个突变都会引起EvC综合症和Weyers颅面骨发育不全(Weyers acrodental dysostosis,WAD)。WAD又称为Curry-Hall综合症(Curry-Hall Syndrome),是一种常染色体显性遗传病,临床主要表现为指甲发育异常、轴后多指、颅面骨发育不全、短肢、身材矮小。WAD与EvC综合症临床表现极其相似,但是症状比EvC综合症稍轻。目前,EVC2基因突变引起疾病的机制还不十分清楚。鸡胚作为经典的胚胎生物学模型被用来研究发育生物学现象已有多年历史,具有周期短、成本低等优势,尤其方便进行显微手术操作。本课题拟利用鸡胚模型研究EVC2基因在早期胚胎发育中的作用及其机制,从而阐明EVC2基因突变的致病机理,为治疗EvC综合征和WAD提供理论依据。首先,我们通过原位杂交检测了EVC2在原肠期的时空表达谱;而后我们通过荧光原位杂交和免疫荧光,在后脑背侧发现了一团共表达EVC2和AP-2α的细胞,AP-2α是颅面神经嵴细胞的Marker,可见EVC2作用于颅面神经嵴细胞;为了进一步研究EVC2对颅面神经嵴细胞的作用,我们利用电穿孔转染的方法过表达和下调EVC2观察其对颅面神经嵴细胞迁移的影响。最后,为了研究EVC2的作用机制,我们表达了EVC2蛋白,拟用CO-IP找出EVC2蛋白作用的靶分子。综上所述,我们通过鸡胚模型,发现EVC2通过作用于早期胚胎中的颅面神经嵴细胞而发挥功能。该发现有助于深入探讨EVC2基因突变的致病机理,从而利于人类对EvC综合征和WAD进行有效预防和治疗。结论:EVC2对早期胚胎发育起重要的作用,它通过作用于颅面神经嵴细胞而发挥功能。
[Abstract]:Ellis-van Creveld syndrome (Ellis-van Creveld Syndrome-EvCor), also known as chondroectodermal dysplasia (chondroectodermal dysplasia), is an autosomal recessive hereditary disease. Cardiac dysplasia. 60% of EvC patients have congenital heart disease. EVC2limbinus LBN) is a kind of integrin, which plays an extremely important role in normal growth and development. Research shows that before birth, it is in the heart, lung, liver, kidney, pancreas, Any mutation in the expression of the .EVC2 gene or EVC gene in a variety of tissues and organs, such as the motor muscle (skeletal muscle), can cause EvC syndrome and Weyers craniofacial dysplasia. Wad, also known as Curry-Hall syndrome Curry-Hall Syndromeg, is an autosomal dominant hereditary disease. The main clinical manifestations were nail dysplasia, posterior fingers, craniofacial dysplasia, short limbs and short stature. WAD was similar to EvC syndrome, but the symptoms were slightly lighter than that of EvC syndrome. At present, the mechanism of EVC2 gene mutation is not well understood. Chicken embryo, as a classical embryonic biological model, has been used to study developmental biological phenomena for many years. It has the advantages of short cycle and low cost, especially for microsurgery. The purpose of this study is to study the role and mechanism of EVC2 gene in early embryonic development by using chicken embryo model, so as to elucidate the pathogenic mechanism of EVC2 gene mutation and provide theoretical basis for the treatment of EvC syndrome and WAD. First, we detected the spatiotemporal expression profiles of EVC2 in the proto intestinal phase by in situ hybridization, and then we used fluorescence in situ hybridization and immunofluorescence. A group of cells co-expressing EVC2 and AP-2 伪 was found in the dorsal side of the posterior brain. AP-2 伪 is the Marker of craniofacial nerve crest cells, and EVC2 acts on the craniofacial nerve crest cells, in order to further study the effect of EVC2 on craniofacial nerve crest cells. We used electroporation transfection method to overexpression and down-regulate EVC2 to observe its effect on craniofacial crest cell migration. Finally, in order to study the mechanism of EVC2, we express the EVC2 protein and use CO-IP to find the target molecule of EVC2 protein. In conclusion, we found that EVC2 functions by acting on craniofacial nerve crest cells in early embryos through chicken embryo model. The findings are helpful to further study the pathogenetic mechanism of EVC2 gene mutation and to effectively prevent and treat EvC syndrome and WAD. ConclusionEVC2 plays an important role in early embryonic development by acting on craniofacial crest cells.
【学位授予单位】：广东药学院
【学位级别】：硕士
【学位授予年份】：2011
【分类号】：R321

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