ADAM10在小鼠颅面部膜内成骨过程中的表达

目的观察解聚素样金属蛋白酶10(A disintegrin and metalloprotease 10,ADAM10)在小鼠颅面部骨骼膜发育过程中的表达变化。方法以野生型C57BL/6小鼠为实验对象,阿辛蓝-茜素红染色显示小鼠膜内成骨区域,结合免疫组织荧光,检测ADAM10在骨组织中的表达。三标免疫荧光观察ADAM10在MC3T3细胞系中亚细胞定位。蛋白免疫印迹检测ADAM10在小鼠出生前后的表达量变化。结果组织阿辛蓝茜素红染色显示小鼠前颅骨、鼻旁软骨、上颌骨、腭板和下颌骨成骨活跃,并结合免疫组织荧光检测,发现ADAM10在小鼠前颌骨、鼻旁软骨、上颌骨、腭板和下颌骨广泛表达,且主要表达在成骨活跃的区域。MC3T3细胞三标免疫荧光检测进一步定位ADAM10广泛分布在胞浆中,在质膜和细胞核附近表达高;其胞核附近的高表达信号与高尔基体共标,提示其可能在高尔基体中加工后至细胞膜发挥功能。同时,蛋白免疫印迹结果证实,ADAM10在小鼠出生前后表达高,成年后表达明显降低。结论 ADAM10广泛表达于小鼠早期颅面部膜内成骨活跃区域,提示其可能调控小鼠颅面部膜内成骨过程。

Expression of ADAM10 During Craniomaxillofacial Intramembranous Ossification of Mice

Objective To investigate the expression of ADAM10 during craniomaxillofacial ossification of mice. Methods Alcian blue and Alizarin red staining were used to detect the intramembranous ossification areas during craniomaxillofacial development of wild-type C57BL/6 mice. Expression of ADAM10 in mice craniomaxillofacial bone was analyzed by immunohistochemistry. The subcellular localization of ADAM10 in MC3T3 was observed by double immunofluorescent staining and the expression of ADAM10 in mice mandibular was determined by western blot. Results Skull, nasal cartilage, maxilla, palate plate and mandibular of mice displayed intramembranous ossification and expression of ADAM10 showed strong singal in these regions, especially the regions undergoing intramembranous ossification. In MC3T3, ADAM10 distributed in cytoplasm, especially in cell membrance and cell nucleus, and the high expression signal of ADAM10 near cell nucleus was co-localized with GM130, a marker of Golgi, indicating ADAM10 decorated in Golgi before transported to the cell membrance. The expression of ADAM10 in mice mandibular was significantly down-regulated when grown up. Conclusion ADAM10 is widely expressed in mice intramembranous ossification regions, providing the evidence of its regulation in intramembranous ossification of mice.