作　　者： (魏安瑶）; (赵鹏辉）; (夏景兰）; (王强）; (杜兴华）;

机构地区： 河北安国中学,河北安国071200

出　　处： 《发育医学电子杂志》 2017年第1期7-13,共7页

摘　　要： 目的探讨miRNA-23a对斑马鱼颅面软骨,尤其是咽区软骨发育过程中神经嵴细胞诱导、迁移和分化的影响。方法通过在胚胎中注射吗啉环修饰的反义核苷酸(antisense morpholino oligonucleotide,MO),抑制胚胎内源性miRNA-23a的活性。阿尔新蓝染色观察敲低mi RNA-23a对斑马鱼颅部软骨发育的影响。整胚原位杂交检测不同发育时期神经嵴细胞特异基因的表达情况。分别在Tg(sox10:EGFP)和Tg(col2a1:h2a-mCherry)转基因胚胎中敲低mi RNA-23a,激光共聚焦显微镜活体检测咽区软骨形成和神经嵴细胞向软骨分化过程中存在的缺陷。结果受精后96小时的敲低miRNA-23a胚胎头部背侧颅骨部分的索旁软骨后段缺失,且头部腹侧咽区软骨中的麦氏软骨、腭方软骨和角舌骨明显变短,基舌骨和角鳃骨缺失。后续的原位杂交实验及共聚焦显微镜拍摄结果表明,敲低miRNA-23a没有影响神经嵴细胞的形成及向头部迁移,但从后脑迁移到咽区的神经嵴细胞明显减少,而且不能分化为成熟的软骨细胞,致使角鳃骨缺失。结论 miRNA-23a对颅神经嵴细胞向咽区迁移并分化为软骨细胞具有重要作用。 Objective To investigate the role of microRNA-23a （miRNA-23a） in the migration and differentiation of cranial neural crest （CNC） cells during craniofacial cartilage development. Methods First, antisense morpholino oligonucleotide against miRNA-23a （23a-MO） was injected into zebrafish embryos to inactivate endogenous miRNA-23a, and alcian blue staining was applied to analyze the developmental defects of craniofacial cartilages in miRNA-23a morphants. Next, the induction and migration of CNC cells in 23aMO injected embryos were examined by whole mount in situ hybridization methods. Furthermore, the effect of miRNA-23a on the pharyngeal migration and chondrogenic differentiation of CNC cells was analyzed in Tg（soxlO：EGFP）;Tg（col2al：h2a-mCherry） double transgenic embryos using confocal laser scanning microscope. Results Knockdown of miRNA-23a led to a severe loss of the posterior part of parachordal cartilages located in dorsal neurocranium. Moreover, ventral pharyngeal skeletal elements, including the Meckel＇ cartilage, ceratohyal cartilage and palatoquadrate, were obviously shortened, and the basihyal and ceratobranchial cartilages were absent in miRNA-23a morphants. The results of subsequent experiments showed that CNC cells were normally developed and migrated into the head regions, but could not migrate from hindbrain to pharyngeal regions and lost their ability to undergo chondrogenic differentiation in miRNA- 23a morphants. Conclusion miRNA-23a is indispensable for the migration and differentiation of CNC cells.

关 键 词： 斑马鱼 神经嵴细胞 颅面软骨