All optical control and monitoring of neural activity

全光学控制和神经活动监测

• 批准号: 8638689
• 负责人: Samarendra Kumar Mohanty
• 金额: $ 17.61万
• 依托单位: UNIVERSITY OF TEXAS ARLINGTON
• 依托单位国家: 美国
• 财政年份: 2013
• 资助国家: 美国
• 起止时间: 2013-09-15 至 2015-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8638689
• 关键词: Animal Model; Cell Culture Techniques; Cells; Clinical Treatment; Controlled Study; Detection; Development; Electric Stimulation; Electrophysiology (science); Electroporation; Embryo; Frequencies; Ganglion Cell Layer; Gene Delivery; Gene Transfer; Genes; Genetic; Goals; Human Activities; Image; In Vitro; Inflammatory Response; Interferometry; Intervention; Label; Lasers; Length; Light; Location; Measures; Methods; Molecular; Monitor; Neurons; Opsin; Optical Coherence Tomography; Optics; Patients; Pattern; Peripheral; Phase; Photoreceptors; Physiologic pulse; Plague; Plasmids; Proteins; Rattus; Reaction; Resolution; Retina; Retinal Ganglion Cells; Retinitis Pigmentosa; Signal Transduction; Source; Staging; System; Techniques; Technology; Tissues; Transfection; Viral; Viral Vector; Width; base; blind; cell type; human disease; in vivo; kidney cell; minimally invasive; nano; nanometer; novel; optogenetics; promoter; public health relevance; relating to nervous system; success

Project Summary: Success of optogenetic intervention of neural activity requires optimization of delivery of genes encoding light sensitive proteins (opsins) to specific cells, and to record the changes in cells and tissue during optogenetic stimulation. The most-commonly used method for delivering opsin(s) is use of viral vector, which is prone to cause unexpected inflammatory responses, immunological reactions, and improper gene integration. Further, the viral methods limit the size of plasmid that can be packaged and delivered and therefore cannot carry multiple opsin-encoding genes or large promoters. Further, in several cases of human diseases such as retinitis pigmentosa (RP) where progressive loss of photoreceptors happens in peripheral retina, it will be useful to localize the expression of the opsins not only in specific cell types, but in a restricted spatial region (peripheral in RP). The first aim of the proposal is to optimize a non-viral delivery method to mitigate the challenges posed by viral delivery. We have recently used focused near-IR ultrafast laser beam method to deliver opsins (ChR2) into spatially-patterned regions of neural tissue (retina). However, this technique needs to be optimized so as to minimize the deleterious effects. Further, it will prove useful to develop a label-free optical technique (in contrast to electrophysiology) to non-invasively evaluate functional activation of optogenetically-sensitized neurons with high spatial resolution and large throughput. Recently, we demonstrated use of Phase-Sensitive Frequency Domain Optical Coherence Tomography (PSFD-OCT) for detection of fluctuations in optogenetically-stimulated cells. PSFD-OCT is a novel technique based on the principles of low-coherence interferometry that can detect displacements of the order of tens of picometers. Because of the low-coherence length of the light, the detected signal has to be within the coherence length of the light source (~10¿m). This enables PSFD-OCT to investigate sub- nanometer changes within a very small region of the tissue volume and is suitable for highly localized detection. The overall aim of this study is to optimize optical delivery of gene encoding opsins, and develop label-free non-invasive optical readout method based on PSFD-OCT to monitor the changes in cortical neurons and tissue resulting from the activation. 1

项目摘要：神经活动光遗传学干预的成功需要优化 将编码光敏蛋白（视蛋白）的基因递送至特定细胞，并记录 光遗传学刺激过程中细胞和组织的变化是最常用的方法。 传递视蛋白是使用病毒载体，容易引起意想不到的炎症 反应、免疫反应和不正确的基因整合此外，病毒方法。 限制了可以包装和递送的质粒的大小，因此不能携带多个 此外，在一些人类疾病中，例如视蛋白编码基因或大启动子。 视网膜色素变性（RP），周边视网膜发生光感受器逐渐丧失， 将有助于定位视蛋白的表达，不仅在特定细胞类型中，而且在 该提案的第一个目标是优化非病毒性空间区域（RP 中的外围）。 我们最近使用了集中传递方法来减轻病毒传递带来的挑战。 近红外超快激光束方法将视蛋白 (ChR2) 传递到空间图案区域 然而，该技术需要进行优化，以尽量减少对神经组织（视网膜）的影响。 此外，开发无标记光学技术将被证明是有用的（相反）。 电生理学）以非侵入性方式评估光遗传学敏化的功能激活 最近，我们演示了具有高空间分辨率和大吞吐量的神经元的使用。 相敏频域光学相干断层扫描 (PSFD-OCT) 用于检测 PSFD-OCT 是一种基于光遗传学刺激细胞的波动的新技术。 低相干干涉测量原理可检测数十个量级的位移 由于光的相干长度较低，检测到的信号必须在皮米以内。 光源的相干长度 (~10¿m) 这使得 PSFD-OCT 能够研究亚光。 纳米变化在组织体积的非常小的区域内，并且适用于高度 本研究的总体目标是优化基因编码的光学传递。 并开发基于 PSFD-OCT 的无标记非侵入性光学读出方法 监测激活引起的皮质神经元和组织的变化。 1