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

本子项目是利用由NIH/NCRR资助的中心赠款提供的资源的众多研究子项目之一。子项目和研究者（PI）可能已经从另一个NIH来源获得了主要资金，因此可以在其他CRISP条目中表示。列出的机构是中心的，不一定是研究者的机构。提出的研究结果将增加我们对氯氧基除草剂毒性作用的了解，并将为果蝇用于人类健康研究提供见解。过去一年的大部分时间都在购买设备和用品，建造一个可行的实验室空间。实验于2005年1月开始。我们在NDSU的合作伙伴Mary Stewart向我们展示了果蝇培养的基础知识，以及将果蝇和胚胎暴露于除草剂和收获果蝇和胚胎的技术。在Specific Aim 1中，我们将建立一个模型来研究果蝇的气管发育。实验1将建立2,4- d、MCPA和溴硝西尼的LC 50。第一次用2,4- d做的实验并不成功，因为所有的苍蝇都死于乙醇作为溶剂。目前正在进行使用不同溶剂（3%乙醇，1%吐温80在水中）的实验。另外，正在测试滤纸上的其他暴露方式。MCPA和溴硝西尼的lc50实验正在进行中。在实验二中，我们研究了模型化合物对果蝇生命周期的影响。目前，我们的实验室正在进行实验，以建立正常苍蝇的详细生命周期。在这些实验中使用了一个保持湿度和温度和昼夜循环的培养箱。为了实现特定目标2，我们已经开始了从果蝇胚胎中提取mRNA和蛋白质的实验，并进行了大量培养果蝇的实验。下一个资助期的目标是完成Specific Aim 1的实验，并开始Aim 2的实验。目前正在招募学生参加2005年夏季的研究。与部落学院建立联系的目的是方便印第安学生从两年制大学过渡到四年制大学，并增加印第安人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.