miR218抑制皮层神经元的辐射迁和胼胝体投射.docxVIP

簿温州医学院硕士学位论文mill一218抑制皮层神经元的辐射迁移和胼胝体投射 簿温州医学院硕士学位论文 mill一218抑制皮层神经元的辐射迁移和胼胝体投射 摘要 目标：microRNAs(miRNAs)是一类非编码的小RNA，uJ．以通过抑制靶基因的翻 译或者引起靶基因mRNA的降解参而与调控许多发育进程。近来许多报道称 miRNA在神经系统发育和神经系统疾病的发生发展过程都承担着重要角色。在 之前的研究中发现，肿瘤生长抑制因子miR一218存神经退行性疾病Huntington 舞蹈病中高表达，并且随着病程的进展表达量进一步增高。所以本文将重点研究 miR．218在大脑皮层投射神经元中的表达及其功能。 方法：首先，我们通过Q．PCR的方法，检测miR一218在小鼠大脑皮层不同发育 期的表达情况；然后，通过原位杂交的方法，确定miR一218在皮层的具体表达位 置；接下来，利用胚胎内基冈转染技术，在体过表达或者抑制miR一218的表达， 研究miR一218在皮层神经元发育过程的功能；另外，通过免疫荧光、RT—PCR的 方法，探讨miR．21 8 uJ．能作用的分子机制；最后，利用双荧光素报告检测的方法， 在体外研究miR一21 8能否调控靶基因Mdgal的翻泽。 结果：miR。218主要表达在小鼠大脑第五皮层。尽管miR一218对神经元的产生没 有影响，在体过表达miR．218后，可以抑制皮层神经冗的迁移和胼胝体投射。而 抑制miR一218的表达，对第五皮层神经元的迁移没有影响。同时，体外实验表明 miR．218叮以抑制Mdgal 3UTR报告载体的荧光素酶活性。 结论：miR．21 8可能通过调控Mdgal的翻译，抑制神经兀的辐射迁移和胼胝体 投射。 关键词：miR一218； 迁移； 胼胝体投射； Mdgal ————————————望堕型韭麴±堂丝．坠窒m．R-218 ————————————望堕型韭麴±堂丝．坠窒 m．R-218 Inhibits Radial Migration and Callosal ProjectiOn of Cortica J Neurons Abstract Purp嗍MjcroRNAs(miRNAs)are small noncoding RNAs that regulate muItipIe deVeIopmenta J processes by repressing translation or triggering degradatiOn 0f the target genes·Recent studies demonstrate that miRNAs play prominent r0Ies in neuraI deVeIOpment and diseases·A tumor suppressor miR一218 was reported to increase in e×pressIon with disease grade in Huntington’s disease of neurodegeneration．In this study，we a．med to elucidate the expression and function of miR．218 in the developing projection neurons of the cerebral cortex Methods：First，the expression of miR一218 in mouse developing cerebral cortex was dete‘m。ned by Q_PCR and／n situ hybridization(ISH)．Second，to characterize the roIe of mIR-218 in cortical development，miR一218 was ectopically expressed or inhibited ‘n the cerebral coRex by／n utero electroporation．Third，Q-PCR and immunohistoche ‘。。。istry were applied to examine the underlying molecular mechanisms of miR一218． At last，in vitro studies with cell cultures were performed to look for miR一218 targets and the targeted sequences． Results：miR一218 was found to be