• 主题
高级搜索  >
历史搜索 清空历史
首页> 外文期刊>Reproductive toxicology >14)C methanol incorporation into DNA and proteins of organogenesis stage mouse embryos in vitro.
【24h】

14)C methanol incorporation into DNA and proteins of organogenesis stage mouse embryos in vitro.

机译：14）C甲醇体外掺入器官形成阶段小鼠胚胎的DNA和蛋白质中。

获取原文
获取原文并翻译 | 示例
           
页面导航
  • 摘要
  • 著录项
  • 相似文献
  • 相关主题

摘要

Methanol (MeOH), a widely used industrial solvent and alternative motor fuel, has been shown to be mutagenic and teratogenic. We have demonstrated that methanol is teratogenic in mice in vivo and causes dysmorphogenesis in cultured organogenesis stage mouse embryos. Although MeOH is a product of endogenous metabolism in the gut and can be found in humans following consumption of various foods, elevated levels of methanol could lead to methylation of cellular macromolecules. DNA methylation has been demonstrated to suppress transcription of fetal genes and may also play an important role in genetic imprinting. Embryonal proteins are also potential targets for methanol-induced methylation. We investigated the potential of administered methanol to incorporate into and/or alter the methylation of embryonal DNA or to affect specific protein methylation. Gestational day 8 CD-1 mouse embryos were grown for 24 h in culture medium (CM) with 0, 4, or 8 mg MeOH + 20 microCi (14)C-MeOH/mL. At the end of the culture period, yolk sacs and embryos were separated for each treatment group. The DNA was purified by cesium chloride gradient centrifugation in the presence of ethidium bromide and (14)C incorporation was determined. Methylation of a selected gene, Hoxc-8, was assessed by using methylation-specific restriction enzymes. The (14)C activity was found superimposed over the DNA-containing fraction, indicating incorporation. DNA from embryos treated with 4 mg MeOH/mL CM gave the highest incorporation of (14)C-MeOH (8 mg/mL was growth inhibiting). Methylation of Hoxc-8 appeared to be increased in embryos treated with 4 mg MeOH/mL CM, but not in embryos treated with 8 mg MeOH/mL. Lack of incorporation of methylation at the higher concentration may be due to the failure of embryos to grow at this concentration of MeOH. The incorporation of (14)C-MeOH into embryo proteins was investigated by polyacrylamide gel electrophoresis (PAGE) and autoradiography. Incorporation of (14)C-MeOH into specific proteins was observed but the labeling specificity was not methanol dose-related. These results indicate that methyl groups from (14)C-MeOH are incorporated into mouse embryo DNA and protein. Our results further suggest that methanol exposure may increase genomic methylation under certain conditions which could lead to altered gene expression.
机译：甲醇（MeOH）是一种广泛使用的工业溶剂和替代发动机燃料，已被证明具有致突变性和致畸性。我们已经证明，甲醇在小鼠体内会致畸，并在培养的器官发生阶段的小鼠胚胎中引起畸形发生。尽管甲醇是肠道内源性代谢的产物，可以在食用各种食物后在人体中发现，但甲醇含量升高可能导致细胞大分子甲基化。 DNA甲基化已被证明可以抑制胎儿基因的转录，并且在基因印记中也可能起重要作用。胚胎蛋白也是甲醇诱导的甲基化的潜在靶标。我们研究了甲醇掺入和/或改变胚胎DNA甲基化或影响特定蛋白质甲基化的潜力。妊娠第8天CD-1小鼠胚胎在含有0、4或8 mg MeOH + 20 microCi（14）C-MeOH / mL的培养基（CM）中生长24小时。培养期结束时，将每个处理组的卵黄囊和胚胎分开。在溴化乙锭存在下，通过氯化铯梯度离心纯化DNA，并确定（14）C掺入。通过使用甲基化特异性限制酶评估所选基因Hoxc-8的甲基化。发现（14）C活性重叠在含DNA的级分上，表明掺入。用4 mg MeOH / mL CM处理的胚胎的DNA掺入最高的（14）C-MeOH（8 mg / mL具有生长抑制作用）。 Hoxc-8的甲基化在用4 mg MeOH / mL CM处理的胚胎中似乎增加，但在用8 mg MeOH / mL CM处理的胚胎中却没有增加。缺乏较高浓度的甲基化可能是由于胚胎无法在此浓度的MeOH下生长所致。通过聚丙烯酰胺凝胶电泳（PAGE）和放射自显影研究（14）C-MeOH掺入胚胎蛋白。观察到将（14）C-MeOH掺入特定蛋白中，但标记特异性与甲醇剂量无关。这些结果表明来自（14）C-MeOH的甲基被掺入小鼠胚胎DNA和蛋白质中。我们的结果进一步表明，在某些条件下，甲醇暴露可能会增加基因组甲基化，这可能导致基因表达改变。

著录项

相似文献

  • 外文文献
  • 中文文献
  • 专利
获取原文

客服邮箱：kefu@zhangqiaokeyan.com

京公网安备：11010802029741号 ICP备案号：京ICP备15016152号-6 六维联合信息科技 (北京) 有限公司©版权所有

  • 客服微信

  • 服务号