EAGER: Optical Control and Reporting of Cystosolic and Organellar pH in situ

所有形式的生命都依赖于生物化学过程，这些过程根据其附近质子的浓度（pH值）而加速或抑制。因此，预测神经细胞内发生的pH变化对神经细胞传递和存储信息的能力具有深远的影响。然而，很难测量或控制活体动物神经细胞中的pH值，因此很难测试pH值变化对神经系统的影响。在这里，研究人员建议使用光学技术，结合遗传编码的蛋白质，来测量和控制果蝇（Drosophila melanogaster）活神经细胞的pH值。荧光蛋白将用于测量pH值，光激活质子泵将用于控制pH值。当部署在同一个细胞中时，这些蛋白质将提供建立pH值动态控制的手段。这些工具对于研究神经细胞在自然发生的活动中pH值波动的程度将是非常宝贵的，并用于测试这些pH变化在维持神经细胞传递和存储信息的能力方面的作用。此外，果蝇神经细胞的这些工具的开发将促进它们在生物科学的其他领域的应用，从真菌学和植物生理学到肾生理学和肿瘤学。通过该项目产生的转基因动物和质粒将直接提供给科学研究界。有关这些工具的性能和应用的信息将通过在科学期刊上发表文章和在科学会议上作专题介绍的方式提供。该项目将直接为MD/PhD研究生提供研究培训，并为博士研究生，本科生，高中生和K-12教师提供培训机会。

All forms of life rely on biochemical processes and these processes are either accelerated or inhibited according to the concentration of protons (pH) in their immediate vicinity. The pH changes that occur within nerve cells are therefore predicted to have a profound influence on the ability of nerve cells to transfer and store information. However, it is very difficult to measure or control pH in the nerve cells of live animals, and as a result the influence of pH changes on the nervous system, is difficult to test. Here, the investigators propose to use optical techniques, combined with genetically-encoded proteins, to measure and control the pH of live nerve cells in fruit flies (Drosophila melanogaster). Fluorescent proteins will be used to measure pH, and light-activated proton pumps will be used to control pH. When deployed in the same cell, these proteins will provide the means for establishing dynamic control of pH. These tools will be invaluable for investigating the extent to which the pH fluctuates in nerve cells during naturally occurring activity, and for testing the role of these pH changes in maintaining the capacity of the nerve cells to transfer and store information. Further, the development of these tools for fruit fly nerve cells will facilitate their application in other fields of the biological sciences, from mycology and plant physiology to renal physiology and oncology. Transgenic animals and plasmids generated through this project will be made available directly to the scientific research community. Information regarding the performance and application of the tools will be made available through publications in scientific journals and presentations at scientific meetings. This project will directly provide research training for an MD/PhD graduate student and training opportunities for PhD graduate students, undergraduate students, high-school students and K-12 teachers.