Perturbation of retinoic acid (RA)-mediated limb development suggests a role for diminished RA signaling in the teratogenesis of ethanol
Perturbation of retinoic acid (RA)-mediated limb development suggests a role for diminished RA signaling in the teratogenesis of ethanol
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DOI：
10.1002/bdra.20385
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发表时间：
2007-09-01
影响因子：
--
通讯作者：
Sulik, Kathleen K.
中科院分区：
文献类型：
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作者：
Johnson, Corey S.;Zucker, Robert M.;Sulik, Kathleen K.
背景：乙醇致畸的机制是乙醇介导的维甲酸（RA）的减少。以肢体缺损为致畸终点的小鼠模型对这一前提进行了研究。方法：C57BL/6J小鼠在妊娠第9天给予乙醇、双硫仑或BMS-189453。妊娠第18天采用阿利新蓝和茜素红染色评估前肢形态。硫酸尼罗蓝或LysoTracker Red （LTR）活体染色鉴定了肢体芽中的细胞死亡。通过共给药和激光扫描共聚焦显微镜ltr染色检查来评估RA预防乙醇诱导细胞死亡的能力。采用原位杂交和qPCR检测处理后的肢体芽的基因表达。结果：乙醇、双硫仑和BMS-189453可导致轴后指畸形、中间指畸形和其他指畸形。在三次暴露后的推定AER中，过量的尼罗河蓝硫酸盐染色是明显的。添加RA可阻止乙醇诱导的LTR染色。原位杂交和qPCR均显示，与对照组相比，暴露于乙醇的胚胎中Shh和Tbx5的表达减少。结论：与长期类风湿性关节炎缺乏的研究相反，急性暴露于功能性类风湿性关节炎拮抗剂导致的肢体缺陷在形态学上与乙醇引起的肢体缺陷相似。类风湿关节炎对乙醇诱导的细胞死亡的拯救以及Shh转录的类似变化进一步表明类风湿关节炎参与了乙醇诱导的肢体畸形。此外，乙醇暴露后肢体发育关键介质的抑制增加了乙醇致病作用的现有知识。
BACKGROUND: A proposed mechanism for ethanol teratogenicity entails ethanol-mediated reductions in retinoic acid (RA). This premise was investigated utilizing a mouse model, with limb reduction defects as the teratogenic end point. METHODS: Ethanol, Disulfiram, or BMS-189453 was administered to C57BL/6J mice on the 9(th) day of pregnancy. Forelimb morphology was assessed on gestation day 18 using Alcian blue and Alizarin red staining. Nile blue sulfate or LysoTracker Red (LTR) vital staining identified cell death in the limb bud. The ability of RA to prevent ethanol-induced cell death was assessed by coadministration followed by laser scanning confocal microscopic examination of LTR-staining. In situ hybridization and qPCR were used to examine gene expression in treated limb buds. RESULTS: Ethanol, Disulfiram, and BMS-189453 resulted in postaxial ectrodactyly, intermediate ectrodactyly, and other digital defects. Excessive Nile blue sulfate staining was evident in the presumptive AER following each of the three exposures. Ethanol-induced LTR staining was prevented by RA supplementation. Both in situ hybridization and qPCR illustrated decreases in Shh and Tbx5 in ethanol-exposed embryos as compared to control. CONCLUSIONS: Contrary to studies of prolonged RA deficiency, acute exposure to functional antagonists of RA results in limb defects that are morphologically similar to those caused by ethanol. The rescue of ethanol-induced cell death by RA and similar changes in Shh transcription further suggest that RA contributes to ethanol-induced limb dysmorphology. Moreover, the repression of key mediators of limb development soon after ethanol exposure adds to the existing knowledge of the pathogenic effects of ethanol.