Quantitation of the physicochemical properties of myelin using Nile Red fluorescence spectroscopy.

IF 4.2 3区 医学 Q2 BIOCHEMISTRY & MOLECULAR BIOLOGY
W Teo, M L Morgan, P K Stys
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引用次数: 0

Abstract

Myelin is a vital structure that is key to rapid saltatory conduction in the central and peripheral nervous systems. Much work has been done over the decades examining the biochemical composition and morphology of myelin at the light and electron microscopic levels. Here we report a method to study myelin based on the fluorescent probe Nile Red. This lipophilic dye readily partitions into live and chemicallyfixed myelin producing bright, well-resolved images of the sheath. Using spectral confocal microscopy, a complete emission spectrum of Nile Red fluorescence can be acquired for each pixel in an image. The solvatochromic properties of Nile Red cause its emission spectrum to change depending on the polarity of its local environment. Therefore, measuring spectral shifts can report subtle changes in the physicochemical properties of myelin. We show differences in myelin polarity in central versus peripheral nervous system and in different regions of central nervous system white matter of the mouse brain, together with developmental and sex variations. This technique is also well suited for measuring subtle changes in myelin properties in live ex vivo white matter specimens. We also demonstrate how light deprivation induces a myelin polarity change in adult mouse optic nerve underscoring a continuing myelin plasticity in response to axonal activity well into adulthood. The Nile Red spectroscopic method allows measurement of subtle physicochemical changes in myelin that can importantly influence its electrical properties and by extension, conduction velocities in axons.

利用尼罗河红荧光光谱定量分析髓鞘的理化特性。
髓鞘是一种重要的结构,是中枢和周围神经系统快速盐传导的关键。几十年来,人们在光镜和电子显微镜下对髓鞘的生化成分和形态进行了大量研究。我们在此报告一种基于荧光探针尼罗河红的髓鞘研究方法。这种亲脂性染料很容易进入活体髓鞘和化学固定的髓鞘,产生明亮、清晰的髓鞘图像。利用光谱共聚焦显微镜,可以获取图像中每个像素的尼罗河红荧光的完整发射光谱。尼罗红的溶解变色特性会导致其发射光谱随局部环境的极性而改变。因此,测量光谱偏移可以报告髓鞘理化性质的微妙变化。我们展示了髓鞘极性在小鼠大脑中枢神经系统与外周神经系统以及中枢神经系统白质不同区域的差异,以及发育和性别差异。这项技术也非常适合测量活体外白质标本中髓鞘性质的微妙变化。我们还展示了光剥夺是如何诱导成年小鼠视神经髓鞘极性变化的,强调了髓鞘在成年后对轴突活动的持续可塑性。尼罗河红光光谱法可以测量髓鞘中微妙的物理化学变化,这些变化会对髓鞘的电特性产生重要影响,进而影响轴突的传导速度。
本文章由计算机程序翻译,如有差异,请以英文原文为准。
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来源期刊
Journal of Neurochemistry
Journal of Neurochemistry 医学-神经科学
CiteScore
9.30
自引率
2.10%
发文量
181
审稿时长
2.2 months
期刊介绍: Journal of Neurochemistry focuses on molecular, cellular and biochemical aspects of the nervous system, the pathogenesis of neurological disorders and the development of disease specific biomarkers. It is devoted to the prompt publication of original findings of the highest scientific priority and value that provide novel mechanistic insights, represent a clear advance over previous studies and have the potential to generate exciting future research.
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