细胞间隙连接蛋白43基因在斑马鱼牙齿早期发育中的表达

发布时间：2018-01-03 21:13

本文关键词：细胞间隙连接蛋白43基因在斑马鱼牙齿早期发育中的表达　出处：《华西口腔医学杂志》2015年04期 　论文类型：期刊论文

【摘要】：目的探讨细胞间隙连接蛋白43(cx43)基因在斑马鱼早期胚胎中的表达模式,为进一步研究其在牙齿发育过程中的功能奠定基础。方法提取受精后72 h的斑马鱼胚胎总RNA,逆转录合成c DNA,特异性扩增cx43基因片段,连接到PGEMT载体并判断连接方向,选择相应的RNA聚合酶体外转录,合成地高辛标记的cx43基因反义m RNA探针。运用全时相胚胎原位杂交技术检测斑马鱼胚胎发育早期cx43基因在牙齿发育部位的表达分布,茜素红染色法比较cx43原位杂交表达区域与咽齿解剖部位的关系。结果获得了cx43基因的反义m RNA探针。cx43基因在斑马鱼受精后各个时期咽弓牙齿相应部位均可见到强棕褐色阳性信号。受精后9 d时cx43原位杂交表达部位与咽齿解剖部位基本重叠一致。结论 cx43基因参与牙胚发育和矿化过程,且主要在矿化晚期发挥关键调控作用。
[Abstract]:Objective to investigate the expression pattern of intercellular connexin 43cx43) gene in zebrafish early embryos. Methods Total RNAs of zebrafish embryos at 72 hours after fertilization were extracted, c DNAs were synthesized by reverse transcription, and cx43 gene fragments were amplified specifically. Ligation to the PGEMT vector and determine the orientation of the ligation, select the corresponding RNA polymerase transcription in vitro. Digoxigenin-labeled antisense m RNA probe of cx43 gene was synthesized and the expression distribution of cx43 gene in the early stage of embryonic development of zebrafish was detected by in situ hybridization. The relationship between the expression region of cx43 in situ hybridization and the anatomical location of pharynx teeth was compared by alizarin red staining. Results the antisense m of cx43 gene was obtained. RNA probe. Cx43 gene can be seen in the corresponding parts of pharyngeal arch teeth of zebrafish at all stages after fertilization. The expression site of cx43 in situ hybridization was consistent with the anatomic site of pharynx tooth. Conclusion cx43 gene is involved in the development and mineralization of tooth germ. And mainly in the late mineralization play a key regulatory role.
【作者单位】：重庆医科大学附属口腔医院牙体牙髓科·口腔疾病与生物医学重庆市重点实验室;
【基金】：国家自然科学基金资助项目(31371473) 重庆市医学重点学科建设基金资助项目(《牙体牙髓病学》)
【分类号】：Q786;R78
【正文快照】： 牙齿发育是一种复杂的生理过程,主要通过上皮和间充质间的相互作用完成,期间涉及多种生长因子和信号途径,在特定的时空调控下共同作用最终成形。细胞间隙连接蛋白43(connexin 43,cx43)直接调控细胞间的连接通讯,进行信息和能量的传递,进而调控细胞的新陈代谢和增殖分化等一系

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