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  

Expression of connexin 43 gene during early dental development in zebra fish

作　　者：徐智云[1] 刘鑫[1] 黄兴[1] 杨德琴[1] 

机构地区：[1]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,重庆400017

出　　处：《华西口腔医学杂志》2015年第4期347-351,共5页West China Journal of Stomatology

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>)

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。Objective This study aims to investigate the expression of connexin 43（cx43） gene during early development in zebra fish and provide a foundation for further research of cx43 gene in tooth development. Methods Total RNA was extracted within 72 h after fertilization of zebra fish embryos and then reversed transcribed to generate the c DNA library. The specific fragments of the cx43 gene were then cloned and connected to the PGEMT vector. After confirming the constructed plasmid, the corresponding RNA polymerase was chosen, and the digoxin-labeled anti-sense m RNA probe of cx43 was synthesized in vitro. The cx43 gene expression of zebra fish indifferent stages was carried out by in situ hybridization. The relationship of the cx43 gene expression and anatomy of the pharyngeal teeth were compared by alizarin red staining. Results The m RNA antisense probe of cx43 was acquired. The positive signal of sepia was observed in the different stages of zebra fish pharyngeal teeth after fertilization. After fertilization for 9 days, the expression site of cx43 in situ hybridization was overlapped in accordance with the anatomical site of the pharyngeal teeth. Conclusion cx43 gene participates in tooth development and mineralization process and plays a crucial role in later mineralization.

关 键 词：细胞间隙连接蛋白43基因 原位杂交 斑马鱼 

分 类 号：Q786[生物学—分子生物学]