Rapid brain-wide optogenetic screening with a noninvasive, dynamically programmable in vivo light source

使用无创、动态可编程体内光源进行快速全脑光遗传学筛查

基本信息

批准号：
10401548
负责人：
Kim Butts-Pauly
金额：
$ 180万
依托单位：
STANFORD UNIVERSITY
依托单位国家：
美国
项目类别：
财政年份：
2022
资助国家：
美国
起止时间：
2022-09-01 至 2025-08-31
项目状态：
未结题

来源：
https://reporter.nih.gov/project-details/10401548
关键词：
Address Amygdaloid structure Animals Behavior Biological Blood Circulation Brain Brain region Cell Nucleus Cerebrum Charge Chemicals Chronic Communities Complex Craniotomy Devices Disease Ensure FOS gene Fiber Focused Ultrasound Half-Life Head Heart Heating Hippocampus (Brain)Image Injections Intravenous Kinetics Light Lighting Location Measures Medial Mediating Memory Methods Modeling Morphine Motor Cortex Mus Neurons Neurosciences Neurosciences Research Opioid Opsin Optics Pathology Pattern Penetration Pharmaceutical Preparations Photometry Photons Prefrontal Cortex Procedures Production Protocols documentation Pump Rapid screening Reporter Resolution Slice Source Specificity Structure Structure of subthalamic nucleus Technology Temperature Testing Time Tissues Training Transducers Transgenic Mice Travel Ultrasonic Transducer Variant Visible Radiation absorption attenuation base biomaterial compatibility biophysical properties brain tissue conditioned place preference cranium design drug seeking behavior excitatory neuron experimental study hemodynamics implantation in vivo interest intravenous injection light emission light intensity millisecond minimally invasive nanoparticle nanophotonic neural circuit neuroregulation optical fiber optical spectra optogenetics photonics preference pressure programs relating to nervous system response sample fixation screening simulation spatiotemporal surface coating tool ultrasound virtual reality

项目摘要

PROJECT SUMMARY/ABSTRACT Optogenetics provides a precise deconstruction of neural circuits by optically manipulating the activity of opsin-expressing neurons with fast temporal responses and neuron-type specificity. A critical challenge of delivering light in the brain for in vivo optogenetics arises from the poor penetration of photons in biological tissue due to the scattering and absorption of light. As a result, in vivo optogenetic stimulation in the deep brain usually requires invasive procedures, such as craniotomy and intracranial implantation of optical fibers. The very invasiveness of these procedures also precludes easy repositioning and volume adjustment of the illuminated region in the same subject. Several strategies have been employed to address the challenges above. Red-shifted and ultrasensitive opsins enable transcranial optogenetics, yet light must travel through all superficial brain tissues with significant power attenuation, heating to other brain structures, potential off-target effects, and an inability to reposition the manipulated brain regions. Tapered optical fibers and reconfigurable nanophotonic circuits enable dynamic illumination across multiple brain regions but still require invasive implantation of photonic devices. Recently the Hong lab demonstrated “sono-optogenetics”, a minimally invasive method for optogenetically manipulating neural activity in vivo via brain-penetrant, focused ultrasound (FUS). Central to this method is mechanoluminescent nanoparticles (MLNPs), which can be delivered intravenously, charged by 400-nm light when passing through superficial vessels, pumped by the heart into cerebral vessels, and then gated by FUS to emit 470-nm light locally for opsin activation, all without exiting the blood circulation. Based on these advances, we propose a rapid brain-wide optogenetic screening approach by producing on-demand light emission at any location or depth in the mouse brain. The long-term objective of this proposal is to noninvasively produce on-demand light emission patterns at any location or depth in the mouse brain for rapid brain-wide optogenetic screening of different brain regions. Specifically, the Hong lab aims to develop a toolbox of MLNPs with distinct emission spectra matching different opsin variants, bright mechanoluminescence, and favorable in-vivo circulation half-life. We will characterize and validate their spectral and biophysical properties by constructing an intravital light source with on-demand emission patterns in the brain of live mice. We then seek to use the FUS-mediated intravital light source to optogenetically stimulate multiple brain regions in the same mouse, thereby fulfilling the dynamic selection of illuminated brain regions. The Butts Pauly lab will facilitate the Hong lab in the design of FUS protocols, ensuring minimal neuromodulatory effects by direct FUS stimulation of the brain. Finally, to demonstrate the unique strengths of this approach in addressing neuroscience challenges, the Hong and Chen labs will screen different brain regions that have been found to store opiate-associated memories in the same mouse brain for dissecting their contributions to the animal’s drug-seeking behavior. The proposed method will provide a valuable tool for the broader neuroscience community to rapidly screen different brain regions underlying a certain behavior or pathology of interest in the same animals.

项目概要/摘要光遗传学通过光学操纵具有快速时间响应和神经元类型特异性的视蛋白表达神经元的活动来提供神经回路的精确解构。在体内光遗传学的大脑中传递光的一个关键挑战来自于光子在生物中的渗透性差。由于光的散射和吸收，体内光遗传学刺激通常需要侵入性手术，例如开颅手术和颅内植入光纤。这些手术的侵入性很大，也妨碍了对同一对象的照明区域进行轻松的重新定位和体积调整，已经采用了几种策略来解决上述挑战。具有显着功率衰减的组织、对其他大脑结构的加热、潜在的脱靶效应以及无法重新定位被操纵的大脑区域使得锥形光纤和可重构的纳米光子电路能够实现动态。最近，洪实验室展示了“声光遗传学”，这是一种通过脑穿透聚焦超声（FUS）来光遗传学操纵体内神经活动的微创方法。方法是机械发光纳米颗粒（MLNP），可以静脉内输送，在穿过浅表血管时用 400 nm 光充电，由心脏泵入大脑血管，然后通过 FUS 门控局部发射 470 nm 光以激活视蛋白，所有这些都无需退出血液循环，基于这些进展，我们提出了一种快速全脑光遗传学筛查方法，通过在任何位置产生按需光发射。该提案的长期目标是在小鼠大脑的任何位置或深度无创地产生按需光发射模式，以对不同的大脑区域进行快速全脑光遗传学筛查。实验室目标开发具有与不同视蛋白变体相匹配的不同发射光谱、明亮的机械发光和有利的体内循环半衰期的 MLNP 工具箱，我们将通过构建具有按需发射模式的活体光源来表征和验证它们的光谱和生物物理特性。然后，我们寻求利用 FUS 介导的活体光源对同一只小鼠的多个大脑区域进行光遗传学刺激，从而实现照明大脑区域的动态选择。 Butts Pauly 实验室将协助 Hong 实验室设计 FUS 方案，通过直接 FUS 刺激大脑确保最小的神经调节作用。最后，为了展示这种方法在应对神经科学挑战方面的独特优势，Hong 和 Chen 实验室将筛选不同的方法。已发现在同一小鼠大脑中存储阿片相关记忆的大脑区域，用于剖析它们对动物寻求药物行为的贡献，所提出的方法将为更广泛的神经科学界提供一个有价值的工具，以快速筛选潜在的不同大脑区域。某些行为或相同动物感兴趣的病理学。