Odaph对牙釉质发育影响的初步研究

目的 利用Runx2条件性敲除小鼠的转基因动物模型初步研究Odaph对牙釉质发育的影响。方法 PCR鉴定转基因小鼠的基因型,实时定量PCR实验方法检测Runx2loxp/loxp鼠和K14-cre-Runx2loxp/loxp基因敲除小鼠下颌磨牙区中Amelx,Ambn,Enam,Amtn,Odam,Scpppq1及Odaph基因mRNA的相对表达含量的改变,最后用免疫组化技术检测Runx2及Odaph在基因敲除小鼠下颌磨牙区的表达情况。结果 实时荧光定量PCR结果显示:与野生型小鼠相比,在出生后10 d基因敲除小鼠中Amelx,Ambn,Enam基因mRNA的相对表达量均上调,Amtn,Odam,Scpppq1及Odaph的表达显著下降;免疫组化染色结果显示:Anti-Runx2在出生后10 d敲除型小鼠成釉细胞胞核中呈阴性表达,Anti-Odaph在出生后10 d野生型小鼠成釉细胞远端呈强阳性表达,然而在敲除型小鼠中表达不明显。结论 Odaph在牙釉质发育过程中具有重要作用。

Preliminary study on the effect of Odaph on enamel development

Objective To investigate the effects of Odaph on enamel development using a transgenic animal model of Runx2 conditional knockout mice.Methods The genotypes of transgenic mice were identified by PCR,and real-time quantitative PCR was used to detect the relative expression of Amelx,Ambn,Enam,Amtn,Odam,Scpppq1 and Odaph mRNA in the mandibular molar region of Runx2loxp/loxp and K14-cre-Runx2loxp/loxp knockout mice.Finally, the expression of Runx2 and Odaph in the mandibular molar of knockout mice was detected by immunohistochemistry.Results Real-time quantitative PCR showed that compared with WT mice,the relative expression levels of Amelx,Ambn,Enam mRNA in PN10 cKO mice were up-regulated, and the expressions of Amtn,Odam,Scpppq1 and Odaph were significantly decreased.The results of immunohistochemical staining showed that anti-Runx2 was negatively expressed in the nucleus of ameloblasts of PN10 null mice,and anti-Odaph was strongly detected in the distal end of ameloblasts of PN10 WT mice.However,in cKO mice, the staining was not very obvious.Conclusion Odaph plays an important role in enamel development.