本文選題：EVC2 + 胚胎��； 參考：《廣東藥學(xué)院》2011年碩士論文

【摘要】：Ellis-van Creveld綜合癥(Ellis-van Creveld Syndrome、EvC)又稱(chēng)為軟骨外胚層發(fā)育不良癥(chondroectodermal dysplasia),是一種常染色體隱性遺傳病,臨床主要表現(xiàn)為軟骨營(yíng)養(yǎng)障礙,多指(趾),外胚層發(fā)育不良,心臟發(fā)育異常。60%的EvC患者患有先天性心臟病。EVC2(limbin,LBN)是一種整合蛋白,它在正常的生長(zhǎng)和發(fā)育過(guò)程中起著極其重要的作用。研究表明:出生前,它在心臟、肺、肝臟、腎臟、胰腺、運(yùn)動(dòng)肌(骨骼肌)等多種組織器官中表達(dá)。EVC2基因或EVC基因任何一個(gè)突變都會(huì)引起EvC綜合癥和Weyers顱面骨發(fā)育不全(Weyers acrodental dysostosis,WAD)。WAD又稱(chēng)為Curry-Hall綜合癥(Curry-Hall Syndrome),是一種常染色體顯性遺傳病,臨床主要表現(xiàn)為指甲發(fā)育異常、軸后多指、顱面骨發(fā)育不全、短肢、身材矮小。WAD與EvC綜合癥臨床表現(xiàn)極其相似,但是癥狀比EvC綜合癥稍輕。目前,EVC2基因突變引起疾病的機(jī)制還不十分清楚。 雞胚作為經(jīng)典的胚胎生物學(xué)模型被用來(lái)研究發(fā)育生物學(xué)現(xiàn)象已有多年歷史,具有周期短、成本低等優(yōu)勢(shì),尤其方便進(jìn)行顯微手術(shù)操作。本課題擬利用雞胚模型研究EVC2基因在早期胚胎發(fā)育中的作用及其機(jī)制,從而闡明EVC2基因突變的致病機(jī)理,為治療EvC綜合征和WAD提供理論依據(jù)。首先,我們通過(guò)原位雜交檢測(cè)了EVC2在原腸期的時(shí)空表達(dá)譜;而后我們通過(guò)熒光原位雜交和免疫熒光,在后腦背側(cè)發(fā)現(xiàn)了一團(tuán)共表達(dá)EVC2和AP-2α的細(xì)胞,AP-2α是顱面神經(jīng)嵴細(xì)胞的Marker,可見(jiàn)EVC2作用于顱面神經(jīng)嵴細(xì)胞;為了進(jìn)一步研究EVC2對(duì)顱面神經(jīng)嵴細(xì)胞的作用,我們利用電穿孔轉(zhuǎn)染的方法過(guò)表達(dá)和下調(diào)EVC2觀察其對(duì)顱面神經(jīng)嵴細(xì)胞遷移的影響。最后,為了研究EVC2的作用機(jī)制,我們表達(dá)了EVC2蛋白,擬用CO-IP找出EVC2蛋白作用的靶分子。 綜上所述,我們通過(guò)雞胚模型,發(fā)現(xiàn)EVC2通過(guò)作用于早期胚胎中的顱面神經(jīng)嵴細(xì)胞而發(fā)揮功能。該發(fā)現(xiàn)有助于深入探討EVC2基因突變的致病機(jī)理,從而利于人類(lèi)對(duì)EvC綜合征和WAD進(jìn)行有效預(yù)防和治療。結(jié)論:EVC2對(duì)早期胚胎發(fā)育起重要的作用,它通過(guò)作用于顱面神經(jīng)嵴細(xì)胞而發(fā)揮功能。
[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 偽 was found in the dorsal side of the posterior brain. AP-2 偽 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.
【學(xué)位授予單位】：廣東藥學(xué)院
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2011
【分類(lèi)號(hào)】：R321

【共引文獻(xiàn)】

相關(guān)博士學(xué)位論文 前1條

1 李海燕;短肢矮小患兒的臨床特征及CTSK和IFT80基因的突變研究[D];中國(guó)醫(yī)科大學(xué);2009年

相關(guān)碩士學(xué)位論文 前1條

1 徐高連;單核苷酸多態(tài)性核酸試紙條檢測(cè)技術(shù)的建立[D];佳木斯大學(xué);2007年

，

本文編號(hào)：1910520