RII Track-2 FEC: The Creation of Next-Generation Tools for Neuroscience - Noninvasive Radioluminescence Approaches to Optogenetics

非技术描述这项以研究基础设施改善Track-2为重点的EPSCoR协作(RII Track-2 FEC)建议是南卡罗来纳州、阿拉巴马州和新墨西哥州的四个机构之间的合作，它们是克莱姆森大学、阿拉巴马州伯明翰大学、新墨西哥大学和南卡罗来纳州大学。该项目的目的是扩大光遗传学实验方法的使用，该方法自2005年引入以来，对神经生物学产生了革命性的影响。这种方法允许实验者通过闪光来激活单个或一组神经元，并具有高水平的空间和时间控制。标准光遗传学的主要局限性之一是可见光无法穿透活组织的深处。在这个项目中，将开发一个系统，允许使用低剂量X射线而不是可见光作为激活信号。该项目包括让学生参与的多种机会，特别是代表人数不足的少数群体的成员。双方已经达成协议，在现有的研究密集型大学的暑期研究项目中，接待来自温斯罗普大学和新墨西哥北部学院的学生，这两所大学为高度多样化的学生群体提供服务。该项目还包括指导初级教师的计划，特别是在提案开发方面。技术描述这个多学科项目包括逐步开发新的实验方法。首先，将产生辐射发光纳米颗粒(RLP)，当暴露在X射线下时会发出光。RLP将经过化学修饰，允许特定的共价结合到在神经元中表达的基因工程膜结合视蛋白上，确保两种成分紧密接近，从而有效地转移光刺激。该系统将在培养细胞中进行测试(有效性和没有不良副作用)，然后是脑切片准备，最后是完整的动物(大鼠和小鼠)。RLP将通过注射到脑脊液中进入动物体内，这些颗粒可以通过脑脊液扩散到大脑中。然后将测试X射线照射对被固定和自由移动的动物行为的影响，以验证运动和听觉皮质神经细胞的成功激活。

Non-Technical DescriptionThis Research Infrastructure Improvement Track-2 Focused EPSCoR Collaboration (RII Track-2 FEC) proposal is a collaboration between four institutions in South Carolina, Alabama, and New Mexico, namely Clemson University, the University of Alabama Birmingham, the University of New Mexico, and the University of South Carolina. The aim of the project is to extend the uses of the experimental method of optogenetics, which, since its introduction in 2005, has had a transformative impact on neurobiology. This method allows experimenters to activate individual neurons or groups of neurons, with high levels of spatial and temporal control, by flashing light on them. One of the main limitations of standard optogenetics is the inability of visible light to penetrate deep within living tissues. In this project, a system will be developed to allow the use of low-dosage X-rays, rather than visible light, as the activating signal. The project includes multiple opportunities to involve students, especially members of under-represented minority groups. Agreements are in place to host students from Winthrop University and Northern New Mexico College, which serve highly diverse student populations, in existing summer research programs at the research-intensive universities. The project also includes plans for mentoring junior faculty, especially in proposal development.Technical DescriptionThis multi-disciplinary project involves step-wise development of novel experimental methods. First, radioluminescent nanoparticles (RLPs) will be produced that emit light when exposed to X-rays. The RLPs will be chemically modified to allow specific covalent attachment to a genetically engineered membrane-bound opsin protein expressed in neurons, ensuring close proximity of the two components for efficient transfer of the light stimulus. The system will be tested (for efficacy and the absence of undesired side effects) in cultured cells, then brain slice preparations, and finally in intact animals (rats and mice). RLPs will be introduced into animals by injection into the cerebrospinal fluid, through which the particles may diffuse into the brain. The effects of X-ray exposure on the behavior of immobilized and freely moving animals will then be tested to verify successful activation of neural cells in the motor and auditory cortexes.