维甲酸诱导鼠磨牙牙胚midkine基因表达的研究

The effects of retinoic acid on the expression of midkine gene in mouse tooth germ tissues in vitro

目的 :探讨RA与midkine (MK)基因表达在牙胚发育过程中的相互关系。方法 :将 16d胎龄的鼠胎下颌第一磨牙牙胚分别置于含不同浓度的外源性RA(分别为 5× 10 -8,5× 10 -7和 5× 10 -6mol/L)的RPMI 16 40半固态培养基表面。培养 2d后分别提取总RNA ,经RT -PCR扩增 2 5个循环 ,采用Southern印迹法检测MKmRNA在牙胚组织中的表达变化。结果 :体外培养 2d后 ,对照组 (无外源性RA )牙胚MKmRNA的表达为弱阳性 ;随培养基中外源性RA浓度的增加 ,牙胚组织中MKmRNA的表达逐渐增强。结论 :MK基因在牙胚发育中的作用可能受RA调控。

AIM: To investigate the effects of RA on the expression of midkine (MK) gene in mouse molar germs. METHODS: The first mandibular molar germs of the 16-day old mouse embryos were separated and cultured on RPMI-1640 semisolid medium containing different concentration of exogenous RA(5×10 -8 , 5×10 -7 and 5×10 -6 mol/L ) for 2 days repectively. After the end of culture, isolating total RNA from cultured tooth germs. Amplication reaction was going on for 25 cycles by RT-PCR. The expression of MK gene in mouse molar germs was investigated by Southern blot. RESULTS: In control medium not containing RA, the expression of MK in mouse tooth germs was weak. With the increase of RA concentration in medium, the expression of MK gene in tooth germs gradully increased. CONCLUSION: This study suggested that the function of MK gene in tooth germ development may be regulated by RA .