EFFECT OF MCPA 2,4 D AND BROMOXYNIL ON LUNG DEVELOPMENT

该子项目是利用NIH/NCRR资助的中心赠款提供的资源的许多研究子项目之一。子项目和研究者（PI）可能从另一个NIH来源获得主要资金，因此可以在其他CRISP条目中表示。所列机构为中心，不一定是研究者所在机构。提出的研究结果将增加我们对氯苯氧基除草剂的毒性效应的理解，并将提供深入了解在使用果蝇对人类健康的研究。过去一年的大部分时间都在购买设备和用品，并建立一个可行的实验室空间。实验于2005年1月开始。我们在NDSU的合作者玛丽斯图尔特向我们展示了果蝇培养的基础知识，以及用于将苍蝇和胚胎暴露于除草剂以及收获苍蝇和胚胎的技术。对于具体目标1，我们将开发一个模型来研究果蝇的气管发育。实验1将确定2，4-D、MCPA和溴苯腈的LC 50。用2，4-D进行的第一次实验并不成功，因为所有的苍蝇都因为使用乙醇作为溶剂而死亡。目前正在进行使用不同溶剂（3%乙醇、1%吐温80水溶液）的实验。此外，还在测试滤纸上的替代暴露。MCPA和溴苯腈的LC 50实验正在进行中。对于实验2，我们研究了模型化合物对果蝇生命周期的影响。目前，我们正在实验室进行实验，以建立正常苍蝇的详细生命周期。在这些实验中使用保持湿度和温度以及昼夜循环的培养箱。具体目标2，我们已经开始实验，以优化mRNA和蛋白质提取果蝇胚胎和培养大量的果蝇。下一个供资期的目标是完成具体目标1的实验并开始目标2的实验。目前正在招募学生参加2005年夏季的研究。与部落学院建立了联系，目的是使美洲原住民学生从两年制学院轻松过渡到四年制学院，并增加美洲原住民对生物医学研究的参与。

This subproject is one of many research subprojects utilizing the resources provided by a Center grant funded by NIH/NCRR. The subproject and investigator (PI) may have received primary funding from another NIH source, and thus could be represented in other CRISP entries. The institution listed is for the Center, which is not necessarily the institution for the investigator. Results of studies proposed will increase our understanding about the toxic effects chlorophenoxy herbicides and will provide insight in the use of Drosophila for studies on human health. Most of the past year was occupied with buying equipment and supplies and building a workable laboratory space. Experiments were begun in January 2005. Our collaborator Mary Stewart at NDSU showed us the basics for Drosophila culture and techniques used for exposure of flies and embryos to herbicides, and harvesting flies and embryos. For Specific Aim 1 we will develop a model to study tracheal development of the Drosophila. Experiment 1 will establish the LC 50 of 2,4-D, MCPA and Bromoxynil. The first experiments done with 2,4-D were not successful because all the flies died due to the ethanol used as a solvent. Experiments using a different solvent (3% ethanol, 1 % Tween 80 in water) are currently underway. Also alternative exposure on filter papers is being tested. LC 50 experiments for MCPA and Bromoxynil are underway. For Experiment 2 we study the effect of the model compounds on the life cycle of the flies. Currently experiments are being done to establish a detailed life cycle in our laboratory for normal flies. An incubator that maintains humidity and temperature and day and night cycle is used in these experiments. For Specific aim 2 we have started experiments to optimize mRNA and protein extraction from Drosophila embryos and of culturing large quantities of Drosophilas. The goals for the next funding period are to finish the experiments of Specific Aim 1 and start the experiments of Aim 2. Currently students are recruited for the research in the summer of 2005. Connections with the tribal college have been made with the goal to easy transition of Native American students from a 2-year college to a 4-year college and increase the participation of Native Americans in biomedical research.