维甲酸诱导腭裂小鼠舌发育异常研究

Research of abnormal tongue development in retinoic acid induced cleft palate mouse

关键词:  维甲酸  舌肌发育  生肌调控因子  腭裂

Key words:  [Key words] 　retinoic acid  tongue myogenesis  myogenic regulatory factors  cleft palate

摘要:

目的　通过检测生肌决定因子基因家族成员Myf5和MyoD在维甲酸诱导腭裂小鼠舌肌发育中的表达，探究维甲酸是否在舌肌发育早期影响其肌向分化。方法　建立维甲酸诱导腭裂小鼠模型，利用免疫组化染色法和实时定量PCR法，检测胚胎13天即舌肌发育早期实验组和对照组小鼠舌肌中Myf5和MyoD的分布和表达水平。结果　免疫组化结果显示Myf5和MyoD在舌肌细胞的细胞核中表达，实验组Myf5和MyoD的蛋白表达均低于对照组,且差异有显著性意义（P<0.01）。实时定量PCR结果显示实验组Myf5和MyoD的mRNA表达水平均低于对照组，且差异有显著性意义（P<0.01）。结论　Myf5和MyoD的异常低表达提示了维甲酸可能直接导致舌肌发育和分化的延迟，进而导致腭裂的发生或使其表型加重。

Abstract:

[Abstract] Objective　To investigate the expressions of the myogenic regulation factors (MRFs) Myf5 and MyoD in tongue myogenesis of cleft palate mice induced by RA, and to explore whether the retinoic acid (RA) affect the myogenic differentiation of tongue in early development stage.Methods　Mouse model of RA-induced cleft palate was set up. Immunohistochemistry and real-time PCR were used to detect the distribution and expression levels of Myf5 and MyoD in tongue of RA induced cleft palate group and wild type at embryonic day13 (E13), the early stage of tongue myogenesis. Results Immunohistochemical results showed that Myf5 and MyoD expressed in the nucleus of tongue muscle cells, and the protein levels of them were down regulated in RA-treated group (P<0.01). Real-time PCR results showed that the mRNA expression level of Myf5 and MyoD also decreased significantly in RA-induced cleft palate group at E13 compared to the wild type (P<0.01). Conclusion　The aberrant low expression of Myf5 and MyoD suggests that the delay development and differentiation of tongue muscle may be induced directly by RA. And this could lead to cleft palate or exacerbate its phenotype of cleft palate.