TERE1基因在小鼠胚胎双前肢正常发育和异常发生中的表达
Expression of TERE1 in Normal Forelimbs Development and Retinoic Acid-induced Short Limb Malformations in Mouse Embryogenesis
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摘要:
目的 观察移行上皮反应基因(TERE1)在小鼠胚胎双前肢正常发育和异常发生中的表达,探讨TERE1在短肢发生中的作用。 方法 ICR小鼠受孕后,将其随机分为实验、对照两组,各64只。于孕第10天(GD10),经口灌胃一次给予实验组孕鼠80 mg/kg的全反式视黄酸(atRA)、对照组孕鼠给予等体积的大豆油,并分别于GD11~GD18取两组胎鼠的双前肢。于GD12取下小鼠胚胎双前肢,以atRA诱导,培养72 h后收获肢。利用实时荧光定量聚合酶链反应方法检测TERE1在各样本中的表达情况。 结果 在整体动物试验中, atRA可诱致小鼠胚胎明显的短肢畸形;在体外试验中, atRA可诱致小鼠胚胎前肢多种骨骼发育异常。TERE1在正常、异常肢及培养肢芽中均有表达。在正常和异常肢中,TERE1的表达模式一致;除GD15外,实验肢中TERE1的表达均低于相同时点正常肢的表达水平。在体外培养肢,各组的表达量随培养时间的延长而增加。实验组在培养24h, atRA各剂量组的表达量均低于对照组(P<0.05);培养48h和72h,在2.5& #215;10-6mol/L~1.0& #215;10-5mol/L剂量范围有随剂量增加表达量增加的趋势,但在高剂量组(2.0& #215;10-5mol/L)表达又下降。 结论 TERE1与小鼠胚胎前肢的发育过程有关, atRA在诱致小鼠前肢异常发生中可引起TERE1 mRNA的表达改变。 Abstract:Objective To observe transitional epithelial response gene1 (TERE1) in normal forelimbs development and retinoic acid-induced short limb malformations in mouse embryogenesis, and to explore the relationship between TERE1 and the short limbs. Methods At gestational day 10 (GD10), the gestational mice of the treatment group were administered with 80 mg/kg all-trans retinoic acid (atRA), and those of the control group were administered with the same volume soybean oil. The forelimbs of all embryos were harvested during GD11-GD18. Mice embryonic forelimbs were explanted on GD12 and harvested after cultured for 24 h, 48 h and 72 h induced by atRA in various concentrations. The expressional abundance of TERE1 in all samples was measured by quantitative real-time polymerase chain reaction (QRT-PCR). Results During the whole animal experiments, short limb malformations of mice embryos were induced apparently by atRA, and a variety of skeletal abnormalities of mice embryo forelimbs were also induced by atRA. TERE1 was expressed in all samples. The expression of TERE1 in the short limbs was lower than that in normal limbs on GD11-GD14 and GD16-GD18, but showed no difference between each other on GD15. In the limbs cultured in vitro, the expression of TERE1 was increased with the cultured times. In the treatment group, the expression that affected by different dose atRA was lower than that of the control group at cultured for 24 h (P < 0.05). At cultured for 24 h and 48 h, the expression of this gene increased with the increasing dose of atRA form 2.5& #215;10-6 mol/L to 1.0& #215;10-5 mol/L, but decreased in the highest dose of atRA (2.0& #215;10-5 mol/L) group. Conclusion TERE1 may play a role during the development of the mice embryo forelimbs, and the mRNA expression of TERE1 changed in the forelimb malformations induced by atRA during mouse embryogenesis.
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