Wireless Optogenetics by relay nano-illuminators

中继纳米照明器的无线光遗传学

• 批准号: 8743294
• 负责人: Gang Han
• 金额: $ 33.5万
• 依托单位: UNIV OF MASSACHUSETTS MED SCH WORCESTER
• 依托单位国家: 美国
• 财政年份: 2013
• 资助国家: 美国
• 起止时间: 2013-09-26 至 2017-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8743294
• 关键词: Address; Animals; Area; Behavior; Behavior Control; Biocompatible; Biological; Brain; Brain region; Cell physiology; Chronic; Combinatorial Synthesis; Development; Diffusion; Disease; Exhibits; Fiber; Fiber Optics; Gene Transfer Techniques; Generations; Genes; Genetic; Goals; Growth; Halorhodopsins; Health; Human; Implant; In Vitro; Individual; Inflammation; Infrared Rays; Injection of therapeutic agent; Laboratories; Lanthanoid Series Elements; Lead; Left; Light; Mediating; Methods; Movement; Neurobiology; Neurons; Neurosciences; Opsin; Optics; Output; Particle Size; Penetration; Performance; Positioning Attribute; Property; Reaction; Resolution; Risk; Scheme; Science; Scientist; Signal Transduction; Slice; Solvents; Source; Surface; Temperature; Testing; Tissues; Transgenic Mice; Variant; Viral; Visible Radiation; Wireless Technology; awake; base; biomaterial compatibility; brain tissue; cranium; density; immunogenicity; in vivo; insight; interest; intravenous injection; microbial; nano; nanocrystal; nanoparticle; neural circuit; neuropsychiatry; novel; optical fiber; optogenetics; photonics; public health relevance; research study; spatiotemporal; tool

DESCRIPTION (provided by applicant): An important step toward better understanding neural circuit function was recently made possible thanks to the breakthrough development of optogenetic tools. In this approach, the microbial opsin genes [most notable, Channelrhodopsin (ChR-2) and halorhodopsin (NpHR)] are expressed in neurons either by viral transduction or transgenesis. Neurons expressing opsin can then be activated or inhibited by light at specific wavelengths. Due to its great spatiotemporal resolution, optogenetics is able to functionally dissect brain circuits, as well as to offer new insights into the causal relationship between brain activity and behavior and, possibly, lead to therapies for neuropsychiatric diseases. However, due to the limited tissue penetration of light at the wavelengths necessary to activate optogenetic constructs, the stimulation of behaving animals has to rely on chronically implanted, fiber-optics or mounted LEDs to deliver light into deep brain tissues. Although this method is very useful and has yielded a wealth of information about brain circuits, stimulation via fiber-optics also has important limitations, particularly in regard to chronic stimulation in awake animals. To address this challenging issue, we propose to develop a wireless optogenetic strategy to remotely activate opsins in vivo using a relay nano- illuminator. This approach is buil upon key technologic advances recently made in our laboratory in lanthanide-doped upconversion nanoparticles (UCNPs), a new generation of nanoparticles with unexpected properties. The most significant advantage of UCNPs is their unnatural inverse excitation and emission profiles; i.e., they are excited using biocompatible, low power, deep tissue-penetrant, near infrared radiation that is effectively converted to a higher energy output emission at various shorter wavelengths, including visible light for activation of opsins. We propose two specific aims. For Aim 1, we will characterize the ability of UNCPs to act as "relay illuminators" in vitro and in vivo. We will initially focus on the first generation of UCNP nanoparticles (CaF2 coated core/shell UCNP) that our preliminary experiments have demonstrated to exhibit robust emission from deep brain tissue. In Aim2, we will develop novel combinatorial synthesis to enhance optogenetic performance of lanthanide-doped UCNPs. This new strategy will overcome many of the limitations of current fiber-optic based approaches, and will enable new applications in both fundamental science and human health.

描述（由申请人提供）：由于光学遗传工具的突破性开发，最近使更好地理解神经回路功能的重要步骤成为可能。在这种方法中，微生物Opsin基因[最值得注意的，通道旋转（CHR-2）和卤代汀（NPHR）]在神经元中通过病毒转导或转基因表达。然后，在特定波长处的光激活或抑制表达opsin的神经元。由于其出色的时空分辨率，光遗传学能够在功能上剖析脑电路，并为大脑之间的因果关系提供新的见解 活动和行为，可能导致神经精神疾病的疗法。但是，由于光线在激活光遗传结构所需的波长下的组织渗透有限，因此行为动物的刺激必须依赖于长期植入的，纤维透视或固定的LED，以将光输送到深脑组织中。尽管该方法非常有用，并且已经产生了有关脑回路的大量信息，但是通过光纤启示的刺激也具有重要的局限性，尤其是在清醒动物的慢性刺激方面。为了解决这个具有挑战性的问题，我们建议制定一种无线光遗传策略，以使用继电器纳米照明剂在体内远程激活Opsins。这种方法是基于最近在我们的兰坦德兴奋剂上转换纳米颗粒（UCNP）的实验室中取得的关键技术进步的，这是具有意外特性的新一代纳米颗粒。 UCNP的最重要优势是它们不自然的反激发和排放曲线。即，使用生物相容性，低功率，深组织渗透剂，靠近红外辐射，它们有效地转化为各种的较高能量输出发射 较短的波长，包括可见光以激活Opsins。我们提出了两个具体目标。对于AIM 1，我们将表征UNCP在体外和体内充当“中继照明器”的能力。我们最初将关注第一代UCNP纳米颗粒（CAF2涂层的核心/壳UCNP），我们的初步实验已证明可以表现出来自深脑组织的强大发射。在AIM2中，我们将开发新型的组合合成，以增强兰烷化UCNP的光遗传学性能。这种新策略将克服当前基于光纤的方法的许多局限性，并将在基本科学和人类健康中启用新应用。