Method for Large-scale Quantitative Analysis of Myelin in Living Animals

摘要 多发性硬化症（MS）是一种中枢神经系统脱髓鞘疾病，影响超过230万人 世界各地的人们。其致病机制尚不清楚，诊断也不早 足以防止进一步的神经退化有证据表明，大脑中的脱髓鞘是一种 MS的重要病变与疾病中的认知缺陷密切相关。然而，临床意义 还没有完全建立，因为皮质髓磷脂不能在体内长时间观察到。这里我们 他建议开发光学方法来研究活体动物的皮层髓鞘。一种新的成像方式将 三次谐波产生显微镜（THGM），我们最近证明了这一点， 周围神经系统完整神经中髓鞘的无标记可视化。强大的成像 将开发适合于推进我们对皮层髓鞘形成的知识的能力。首先，THGM将是 在活体MS动物模型中验证用于探测皮质脱髓鞘和髓鞘再生。二、波前 将实施对照以促进小鼠深部脑中皮质髓鞘的大规模THGM成像。 第三，超分辨率THGM形态测量将通过图像恢复和概率建模来实现。 成功完成后，该项目将开辟基础科学研究的新领域， 转化医学，以阐明轴突-胶质细胞相互作用的皮质脱髓鞘， 髓鞘再生

Abstract Multiple sclerosis (MS) is a demyelinating disease of the central nervous system affecting more than 2.3 million people worldwide. The pathogenic mechanism is not well understood and the diagnosis does not occur early enough to prevent further neurodegeneration. Evidence suggests that demyelination in the cerebrum is an important lesion of MS correlating well with cognitive defects in the disease. However, the clinical significance has not been fully established because cortical myelin cannot be observed in vivo over a long period. Here we propose to develop optical methods for studying cortical myelin in living animals. A new imaging modality will be employed, namely third-harmonic generation microscopy (THGM), which we recently demonstrated for label-free visualization of myelin in intact nerves of the peripheral nervous system. Powerful imaging capabilities will be developed suitable for advancing our knowledge of cortical myelination. First, THGM will be validated for probing cortical demyelination and remyelination in living MS animal models. Second, wavefront control will be implemented to facilitate large-scale THGM imaging of cortical myelin in deep mouse brain. Third, super-resolution THGM morphometry will be achieved by image restoration and probabilistic modeling. Upon successful completion, this project will open a new field of investigations in basic science as well as translational medicine to elucidate the axon-glial cell interaction underlying cortical demyelination and remyelination.