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，而这些过程的加速或抑制取决于它们附近的质子浓度（pH）。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有深远的影响。然而，测量或控制活体动物神经细胞中的pH值是非常困难的，因此pH值变化对神经系统的影响是难以测试的。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。荧光蛋白将被用于测量pH值,和光驱动质子泵将用于控制博士当部署在同一单元格中,这些蛋白质会提供博士的方法建立动态控制这些工具对于调查的程度都是无价的pH值波动在神经细胞自然发生的活动,和测试这些pH值变化的角色在维持神经细胞的容量,传输和存储信息。此外，这些果蝇神经细胞工具的发展将促进它们在其他生物科学领域的应用，从真菌学和植物生理学到肾脏生理学和肿瘤学。通过这个项目产生的转基因动物和质粒将直接提供给科学研究界。将通过科学期刊上的出版物和在科学会议上的介绍提供有关这些工具的性能和应用的信息。该项目将直接为一名博士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.