摘要
目的:应用蛋白质组学方法分析间充质细胞在牙齿发育早期不同阶段的蛋白质差异表达情况,从蛋白水平进一步了解牙齿发育早期的分子机制。方法:制备C57胎鼠起始期(E11.5)、蕾状期(E13.5)冰冻切片,通过激光捕获显微切割技术(LCM)获取两个时期下颌第一磨牙牙胚间充质细胞,采用双向凝胶电泳方法分离蛋白质,比较分析凝胶图像,通过质谱及生物信息学分析鉴定差异蛋白。结果:激光捕获了较纯的目的细胞,得到双向凝胶电泳图谱,比较分析发现E13.5组8个蛋白点明显上调,E11.5组5个蛋白点发生明显上调。质谱分析初步鉴定了蛋白质种类。结论:LCM技术能较好地获取较为均一的目的细胞,并发现牙胚发育起始期与蕾状期间充质细胞蛋白质表达存在差异,为牙胚早期发生的分子机制研究提供一种新的方法。
Objective：To investigate the differential expression of proteins in the mesenchymal cells at the early stages of tooth development by proteomics, so as to learn about the mechanisms of tooth development on the protein level. Method：The frozen sections of tooth germs at initiation stage （E11.5）, budding stage （E13.5） in C57 mouse embryos were prepared and the mesenehymal cells of the mandibular first molar in such stages were obtained by LCM. Proteins were separated by Two dimensional polyaerylamide gel electrophoresis （2-DE） technique,and the images of polyacrylamide gels were analyzed. Mass-spectrum identification and bioinformatics analysis was used to study the different proteins. Result： The pure mesenchymal cells were obtained by LCM. Analyzed the image of 2-DE gels, there were 8 prorein spots that were increased markedly at the budding stage （E13.5） ： there were 5 prorein spots that were increased markedly at the initiation stage （E11.5）. They were identified by mass spectrographic analysis. Conclusion： Laser capture microdissection can be applied to successfully obtain pure objective ceils and the differences we have found can provide a new method to study the mechanisms of tooth development.
出处
《临床口腔医学杂志》
2008年第5期263-266,共4页
Journal of Clinical Stomatology
基金
国家自然科学基金资助项目(30400501)
