Visualization of In Vivo Myelin Architecture Using Nonlinear Microscopy

使用非线性显微镜可视化体内髓磷脂结构

• 批准号: 8478142
• 负责人: Hyungsik Lim
• 金额: $ 14.76万
• 依托单位: HUNTER COLLEGE
• 依托单位国家: 美国
• 财政年份: 2012
• 资助国家: 美国
• 起止时间: 2012-06-01 至 2015-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8478142
• 关键词: Algorithms; Animal Model; Animals; Architecture; Award; Axon; Biochemical; Biological Assay; Biophysics; Birefringence; Caliber; Cell Survival; Cells; Charcot-Marie-Tooth Disease; Clinical; Coculture Techniques; Complex; Degenerative Disorder; Demyelinations; Dependence; Development; Diagnostic; Disease; Dyes; Electron Microscopy; Exhibits; Fiber; Generations; Goals; Health; Image; Imagery; In Vitro; Investigation; Label; Lasers; Life; Lighting; Longitudinal Studies; Maintenance; Measures; Membrane; Mentors; Methodology; Methods; Metric; Microscopy; Minor; Molecular; Morphology; Multiple Sclerosis; Myelin; Myelin Sheath; Natural regeneration; Nerve; Nervous System Physiology; Nervous system structure; Neural Conduction; Neuroglia; Neurons; Optics; Pilot Projects; Play; Preparation; Process; Property; Research; Research Personnel; Resolution; Role; Route; Safety; Schwann Cells; Signal Transduction; Specimen; Staining method; Stains; Structure; System; Techniques; Technology; Testing; Thick; Time; Tissues; Training; Visible Radiation; Writing; base; clinically relevant; hereditary neuropathy; imaging modality; improved; in vivo; indexing; innovation; interest; irradiation; light microscopy; light transmission; meter; morphometry; myelination; nervous system disorder; novel; remyelination; repaired; research study; sample fixation; stem

DESCRIPTION (provided by applicant): This revision of an application for a SC2 Pilot Project Award is from young investigator who proposes to develop a non-invasive method for imaging neurons in tissue at sufficient resolution to study myelin structure. If successful, this research could potentially have a major impact on the study of neuro-regeneration and neuro-degenerative diseases. The reviewers really liked this well-written application. Its strengths include the novelty of the imaging method, the project's significance and potential impact, the outstanding qualifications of the PI and his mentors, and the strong development and mentoring plans. Additional strengths were the innovative approach, the well thought out aims, and the already developed imaging system for the studies. A few minor weaknesses were also noted. These included inadequate discussion of relevant developments in the imaging field, of the likelihood that sufficient resolution can be attained, and of the limits of light transmission in tissues. It was felt that te PI had responded well to most of the criticisms of the previous reviewers. Overall, the review panel felt that this is an excellent application from a well-trained young investigator. Although the project is somewhat risky, the potential impact of the results is significant. The proposed research is highly suited to the SC2 mechanism, and the excellent mentoring plan suggests that the PI will significantly benefit from an SC2 Award. ABSTRACT: Myelin is a specialized membrane around axon, which does not only play a significant role in neuronal signaling but is also implicated in several clinical conditions, such s multiple sclerosis and Charcot-Marie- Tooth disease. The complex interaction between axon and glial cells is the crucial mechanism that maintains an optimal level of the myelin thickness with respect to the axon diameter, or g-ratio. However, the dynamic process of myelin formation is not well understood partly because of the lack of technology for measuring the myelin thickness in vivo. Here we propose to develop a non-invasive, imaging-based methodology to determine the thickness of myelin and the g-ratio without exogenous labeling. Our method relies on a nonlinear optical signal generated around the lamellae of myelin. Unlike previous methods, our novel technique will provide micrometer-resolution structural information in live cells and tissue, therefore suited for longitudinal studies of the dynamics of myelination, demyelination, and remyelination. We will undertake appropriate initial experiments in order to validate the correlation between the new method and the myelin structure. First we will test the sensitivity of our imaging using a myelinating co-culture system. Secondly, we will study the polarization-sensitivity as a route to identify the myelin domains. Finally, we will investigate the correlation between our metric and g-ratio using a developing animal model. We anticipate that our proposed research will facilitate in vivo studies of the dynamic interaction between glial cells an axon with minimum animal sacrifice. Moreover, our development may also have a clinical relevance as a diagnostic for demyelination diseases.

描述（由申请人提供）：SC2 试点项目奖申请的修订来自年轻的研究人员，他们提议开发一种非侵入性方法，以足够的分辨率对组织中的神经元进行成像，以研究髓磷脂结构。 如果这项研究成功的话 可能对神经再生和神经退行性疾病的研究产生重大影响。 审稿人非常喜欢这个写得好的应用程序。 它的优势包括新颖性 成像方法、项目的意义和潜在影响、PI 及其导师的杰出资质、以及强有力的发展和指导计划。 其他优势包括创新的方法、深思熟虑的目标以及已经开发的研究成像系统。 还指出了一些小弱点。 其中包括对成像领域相关发展、获得足够分辨率的可能性以及组织中光传输限制的讨论不充分。 人们认为，te PI 对之前审稿人的大部分批评做出了很好的回应。 总的来说，审查小组认为这是一位训练有素的年轻研究者的出色申请。 尽管该项目存在一定风险，但其结果的潜在影响是巨大的。 拟议的研究非常适合 SC2 机制，出色的指导计划表明 PI 将从 SC2 奖中受益匪浅。 摘要：髓磷脂是轴突周围的一种特殊膜，它不仅在神经元信号传导中发挥重要作用，而且还与多种临床疾病有关，例如多发性硬化症和腓骨肌萎缩症。轴突和神经胶质细胞之间复杂的相互作用是维持髓磷脂厚度相对于轴突直径或 g 比的最佳水平的关键机制。然而，由于缺乏体内测量髓磷脂厚度的技术，髓磷脂形成的动态过程尚不清楚。在这里，我们建议开发一种非侵入性、基于成像的方法来确定髓磷脂的厚度和 g 比，而无需外源标记。我们的方法依赖于髓磷脂片层周围产生的非线性光信号。与以前的方法不同，我们的新技术将提供活细胞和组织中微米分辨率的结构信息，因此适合髓鞘形成、脱髓鞘和髓鞘再生动力学的纵向研究。我们将进行适当的初步实验，以验证新方法与髓磷脂结构之间的相关性。首先，我们将使用髓鞘共培养系统测试成像的灵敏度。其次，我们将研究偏振敏感性作为识别髓磷脂域的途径。最后，我们将研究相关性 使用正在开发的动物模型来比较我们的公制和 g 比。我们预计，我们提出的研究将有助于以最小的动物牺牲对神经胶质细胞与轴突之间的动态相互作用进行体内研究。此外，我们的开发还可能具有作为脱髓鞘疾病诊断的临床意义。