7-脱氢胆固醇还原酶在腭突发育中的表达

7-dehydrocholestererol reductase expression in the development stages of palatal shelves

目的:观察7-脱氢胆固醇还原酶(Dhcr7)基因在小鼠胚胎腭突发育中表达量的变化。方法:在体视显微镜下取E13.5、E14.5和E15.5的小鼠胚胎的腭突,免疫组织化学(IHC)观察Dhcr7蛋白在腭突中的表达定位,逆转录式聚合酶链式反应(RT-PCR)法检测Dhcr7 mRNA的表达变化,Western blot法检测蛋白的表达变化。结果:Dhcr7主要表达在胚胎腭突间充质细胞(EPM);E13.5、E14.5和E15.5 mRNA相对表达量(Dhcr7/β-actin)分别是0.692±0.084、0.595±0.051和0.369±0.061;E13.5与E14.5比较,P>0.05;E13.5、E14.5分别与E15.5比较,P<0.01;蛋白相对表达量(Dhcr7/GAPDH)分别是0.857±0.008、0.840±0.005和0.300±0.007,E13.5与E14.5相比,P>0.05;E15.5分别与E13.5和E14.5相比P<0.01。结论:Dhcr7在腭突发育中的关键阶段有表达,并且表达量与其生长发育有密切关系。

Objective： To study Dher7 gene expression in the development stages of mouse embryo palate. Methods： Palate shelves of mouse embryos were obtained on the day of El3.5, El4.5 and El5.5 respectively under stereoscope, the location of Dh- cr7 protein expression in the palatal shelf was observed by immunohistochemistry（ IHC）, the quantification of Dhcr7 mRNA and pro- tein expression was examined by PCR and Western blot respectively. Results： Dhcr7 mainly expressed in the embryo palatal mesen- chymal cells. The relative mRNA expression（Dhcr7/β-actin） at El3.5, El4.5 and El5.5 was O. 692± 0. 084, 0. 595 ±O. 051 and O. 369 ± O. 061 respectively, El5.5 vs El3.5 or El4.5, P 〈 O. 01, El4.5 vs El3.5, P 〉 0.05. The relative expression of protein （Dhcr7/GAPDH） at El3.5, El4.5 and El5.5 was O. 857 ±0. 008, 0. 840 ± 0. 005 and 0. 300 ± O. 007 respectively, El5.5 vs El3.5 or El4.5 P 〈0.01, El4.5 vs El3.5 P 〉0.05. Conclusion： Dhcr7 is expressed in the important stages of the palatal shelf development, and the expression quantity was closely related to the development of palate.