EAGER: Development of a Fluorescent Reporter for Protein-Membrane Interactions

脑细胞中蛋白质与细胞膜的相互作用是大脑工作的基础。这些相互作用也是帕金森氏症等各种神经系统疾病的一个促成因素。该项目涉及开发一种成像方法来研究脑细胞中的蛋白质-膜相互作用。这项研究将有助于研究帕金森氏症的潜在病因，帕金森氏症可能是由一种叫做α -突触核蛋白的蛋白质聚集引起的。然后，成团的蛋白质在整个大脑中从一个脑细胞扩散到另一个脑细胞。尽管α -突触核蛋白聚集过程被认为在帕金森病中起作用，但这种聚集如何发生尚不完全清楚。一种可能的机制涉及-突触核蛋白与脑细胞某些膜的相互作用。随着所提出的成像方法的发展，研究活脑细胞的整个过程将成为可能，从而进一步了解帕金森病。该项目涉及两名研究生。此外，针对七年级数学学生的互动学习模块将在当地一所学校展示。该项目正在为使用系留荧光共振能量转移报告器的成像方案奠定基础，该方案允许对活的、培养的神经元中膜诱导的α -突触核蛋白聚集进行询问。最终目标是研究参与内溶酶体运输的细胞内脂质囊泡，以及突触囊泡，以解决帕金森病活神经元模型中哪些类型的膜结合实体作为α -突触核蛋白结合和聚集位点的关键神经科学问题。为了实现这一研究方向，该项目有两个目标：(i)实现一种监测活神经元中α -突触核蛋白-脂质膜关联的方法，以及（ii）开发一种监测活神经元中膜介导的α -突触核蛋白聚集的技术。这种监测α -突触核蛋白聚集和朊病毒样扩散的光学传感方法的成功开发将使人们能够理解帕金森病，并将其扩展到动物模型研究中。在更普遍的层面上，这个项目可能对理解各种人类疾病、蛋白质-膜相互作用和脑功能的分子基础产生广泛的影响。该奖项反映了美国国家科学基金会的法定使命，并通过使用基金会的知识价值和更广泛的影响审查标准进行评估，被认为值得支持。

The interaction of proteins with membranes in brain cells is fundamental to how the brain works. These interactions are also a contributing factor to various neurologic diseases such as Parkinson’s disease. This project involves the development of an imaging method to study protein-membrane interactions in brain cells. This research will enable studies of the underlying cause of Parkinson’s disease, which may result from the clumping of a protein called alpha-synuclein. The clumped protein then spreads from brain cell to brain cell throughout the brain. Although the alpha-synuclein clumping process is believed to play a role in Parkinson’s disease, how this clumping occurs is not fully understood. One possible mechanism involves interactions of alpha-synuclein with certain membranes in brain cells. With the development of the proposed imaging method, it will become possible to study the entire process in live brain cells and hence develop further understanding of Parkinson’s disease. The project involves two graduate students. In addition, an interactive learning module for seventh grade math students will be presented at a local school.This project is laying the groundwork for an imaging scheme using tethered fluorescence resonance energy transfer reporters to allow interrogation of membrane-induced alpha-synuclein aggregation in live, cultured neurons. The ultimate goal is to study intracellular lipid vesicles involved in endolysosomal transport, as well as synaptic vesicles, to address the key neuroscience question of which types of membrane-bound entities serve as sites of alpha-synuclein binding and aggregation in live neuron models of Parkinson’s disease. To enable this line of research, this project has two objectives: (i) realizing a method to monitor alpha-synuclein -lipid membrane association in live neurons, and (ii) developing a technique to monitor membrane-mediated alpha-synuclein aggregation in live neurons. Success in the development of this optical sensing method to monitor alpha-synuclein aggregation and prion-like spread will enable understanding of Parkinson’s disease that could be extended to animal model investigations. At a more general level, this project could have a broad impact on the understanding of a variety of human diseases, protein-membrane interactions, and the molecular underpinnings of brain function.This award reflects NSF's statutory mission and has been deemed worthy of support through evaluation using the Foundation's intellectual merit and broader impacts review criteria.