F-actin结合蛋白在维甲酸诱导的舌肌发育不良中的作用及机制研究

项目名称： F-actin结合蛋白在维甲酸诱导的舌肌发育不良中的作用及机制研究

项目编号： No.81500827

项目类型： 青年科学基金项目

立项/批准年度： 2016

项目学科： 医药、卫生

项目作者： 丛蔚

作者单位： 大连医科大学

项目金额： 18万元

中文摘要： 舌的正常发育对口颌系统的发育至关重要，舌发育异常与多种口颌畸形密切相关，但目前舌发育异常的病因和机制不清。本课题组前期研究发现维甲酸可导致舌肌发育不良；基因芯片结果显示，与肌肉发育及发育异常密切相关的细胞质骨架F-actin结合蛋白Rai14、DTNA及DMD在维甲酸诱导的舌畸形中表达异常，提示其与舌发育异常相关。本课题拟利用维甲酸诱导的小鼠舌畸形模型，采用免疫荧光染色等方法研究DMD，DTNA和Rai14时空表达及与增殖和分化的相关性；体外采用抑制剂、siRNA干扰或重组蛋白在原代培养细胞及体外培养的器官进行丧失性及获得性功能研究，通过检测蛋白亚细胞定位、细胞超微结构、F-actin动力学等揭示RA与DMD、DTNA、Rai14对舌肌发育的影响及相互作用的分子机制。该研究将有助于揭示舌发育及发育异常的病因及分子机制，为舌发育异常等口颌畸形的早期诊断、预防和治疗提供理论依据。

中文关键词： 舌肌发育不良；维甲酸；DTNA；DMD；；F-actin

英文摘要： Proper tongue development is crucial to the maxillofacial system, and tongue abnormalities are correlated with maxillofacial malformations of human, until now, the pathogenesis and molecular details of abnormal development in tongue remain poorly understood. In our current research, we find that pregnant mice treated with retinoic acid (RA) produce embryos with tongue dysplasia; the analysis of DNA microarray showed that F-actin binding proteins Rai14, DMD and DTNA that are related to myogenesis and myogenic abnormality, are all dysregulated in RA induced tongue malformations, this indicated that Rai14, DMD and DTNA are involved in RA induced tongue dysplasia. On this basis, the project will firstly determine the spatial-temporal dynamic changes of DTNA, DMD and Rai14, and their correlation with proliferation and differentiation during tongue development using immunofluorescence staining. Furthermore, by the loss of function and gain of function studies using inhibitors, siRNA interference or recombinant proteins in primary cell culture of tongue myoblasts and tongue organ culture system, the changes in subcellular localization of protein, F-actin density and distribution, as well as ultrastructural analysis of tongue myoblasts will be explored. Then the interaction of RA, DMD，DTNA and Rai14, and how this network regulates tongue myogenesis will be well illustrated. This research provides a further realization to the pathogenesis and mechanisms of tongue abnormal development and tongue myogenesis, and provides a theoretical basis for the early diagnosis, prevention and clinical treatment of ralated diseases.

英文关键词： Tongue dysplasia;Retinoic acid ;DTNA;DMD;F-actin