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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.

项目总结/摘要光遗传学通过光学操纵具有快速时间反应和神经元类型特异性的视蛋白表达神经元的活动，提供了神经回路的精确解构。体内光遗传学在大脑中传递光的关键挑战来自于由于光的散射和吸收而导致的光子在生物组织中的不良穿透。因此，深部脑中的体内光遗传学刺激通常需要侵入性程序，例如开颅术和颅内植入光纤。这些过程的侵入性也妨碍了在同一对象中的照明区域的容易的重新定位和体积调节。为应对上述挑战，采取了若干战略。红移和超灵敏的视蛋白使经颅光遗传学成为可能，但光必须穿过所有浅表脑组织，具有显著的功率衰减，对其他脑结构加热，潜在的脱靶效应，并且无法重新定位被操纵的脑区域。锥形光纤和可重新配置的纳米光子电路能够在多个大脑区域实现动态照明，但仍然需要光子设备的侵入性植入。最近，Hong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）在体内光遗传学操纵神经活动的微创方法。这种方法的核心是机械发光纳米颗粒（MLNP），它可以通过静脉内输送，在通过浅表血管时由400 nm的光充电，由心脏泵入脑血管，然后由FUS门控以局部发射470 nm的光用于视蛋白激活，所有这些都不离开血液循环。基于这些进展，我们提出了一种快速的全脑光遗传学筛选方法，通过在小鼠大脑的任何位置或深度产生按需光发射。该提案的长期目标是在小鼠大脑中的任何位置或深度非侵入性地产生按需光发射模式，用于不同大脑区域的快速全脑光遗传学筛选。具体来说，Hong实验室的目标是开发一个MLNP工具箱，该工具箱具有与不同视蛋白变体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。我们将通过在活小鼠大脑中构建具有按需发射模式的活体光源来表征和验证其光谱和生物物理特性。然后，我们试图使用FUS介导的活体光源来光遗传学地刺激同一小鼠的多个脑区域，从而实现对照明脑区域的动态选择。Butts Pauly实验室将协助Hong实验室设计FUS协议，确保通过直接FUS刺激大脑来实现最小的神经调节作用。最后，为了证明这种方法在应对神经科学挑战方面的独特优势，Hong和Chen实验室将筛选在同一小鼠大脑中存储阿片类药物相关记忆的不同大脑区域，以剖析它们对动物药物寻求行为的贡献。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，以快速筛选相同动物中某种行为或病理学的不同大脑区域。