利用超分辨率显微镜识别生长锥中的 Z 轴丝状体

IF 4.2 3区 医学 Q2 BIOCHEMISTRY & MOLECULAR BIOLOGY Journal of Neurochemistry Pub Date : 2024-07-01 DOI:10.1111/jnc.16162
Motohiro Nozumi, Yuta Sato, Miyako Nishiyama-Usuda, Michihiro Igarashi
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引用次数: 0

摘要

生长锥是延伸轴突的高度运动尖端,对神经网络的形成至关重要。三维结构照明显微镜是一种超分辨率光显微镜,其分辨率克服了传统光显微镜的光学衍射限制(约 200 nm),非常适合研究细胞内事件的分子动力学。利用这种技术,我们在生长锥内发现了一种沿 Z 轴分布的新型丝状体("Z-丝状体")。Z-丝状体通常沿轴突生长方向分布,不附着于基质,自发突起而无微管侵入,其寿命比传统丝状体短得多。Z-丝状体的形成和动态受肌动蛋白调节蛋白的调控,如血管扩张剂刺激的磷蛋白、法斯金和cofilin。色素辅助激光使cofilin失活可诱导z-filopodia快速周转。轴突导向受体神经蛋白-1集中在z-纤丝体中,并与z-纤丝体一起运输,而其配体semaphorin-3A则选择性地与z-纤丝体结合。与z-filopodia相关的膜域也很特化,类似于脂质筏,其行为与神经蛋白-1密切相关。结果表明,z-filopodia具有独特的周转特性,与xy-filopodia不同,它不能作为轴突延伸的力生成结构。
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Identification of z-axis filopodia in growth cones using super-resolution microscopy

A growth cone is a highly motile tip of an extending axon that is crucial for neural network formation. Three-dimensional-structured illumination microscopy, a type of super-resolution light microscopy with a resolution that overcomes the optical diffraction limitation (ca. 200 nm) of conventional light microscopy, is well suited for studying the molecular dynamics of intracellular events. Using this technique, we discovered a novel type of filopodia distributed along the z-axis (“z-filopodia”) within the growth cone. Z-filopodia were typically oriented in the direction of axon growth, not attached to the substratum, protruded spontaneously without microtubule invasion, and had a lifetime that was considerably shorter than that of conventional filopodia. Z-filopodia formation and dynamics were regulated by actin-regulatory proteins, such as vasodilator-stimulated phosphoprotein, fascin, and cofilin. Chromophore-assisted laser inactivation of cofilin induced the rapid turnover of z-filopodia. An axon guidance receptor, neuropilin-1, was concentrated in z-filopodia and was transported together with them, whereas its ligand, semaphorin-3A, was selectively bound to them. Membrane domains associated with z-filopodia were also specialized and resembled those of lipid rafts, and their behaviors were closely related to those of neuropilin-1. The results suggest that z-filopodia have unique turnover properties, and unlike xy-filopodia, do not function as force-generating structures for axon extension.

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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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