High-Resolution Fluorescence Imaging Combined With Computer Simulations to Quantitate Surface Dynamics and Nanoscale Organization of Neuroligin-1 at Synapses.

IF 4.6 Q2 MATERIALS SCIENCE, BIOMATERIALS
ACS Applied Bio Materials Pub Date : 2022-04-25 eCollection Date: 2022-01-01 DOI:10.3389/fnsyn.2022.835427
Matthieu Lagardère, Adèle Drouet, Matthieu Sainlos, Olivier Thoumine
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引用次数: 0

Abstract

Neuroligins (NLGNs) form a family of cell adhesion molecules implicated in synapse development, but the mechanisms that retain these proteins at synapses are still incompletely understood. Recent studies indicate that surface-associated NLGN1 is diffusionally trapped at synapses, where it interacts with quasi-static scaffolding elements of the post-synaptic density. Whereas single molecule tracking reveals rapid diffusion and transient immobilization of NLGN1 at synapses within seconds, fluorescence recovery after photobleaching experiments indicate instead a long-term turnover of NLGN1 at synapse, in the hour time range. To gain insight into the mechanisms supporting NLGN1 anchorage at post-synapses and try to reconcile those experimental paradigms, we quantitatively analyzed here live-cell and super-resolution imaging experiments performed on NLGN1 using a newly released simulator of membrane protein dynamics for fluorescence microscopy, FluoSim. Based on a small set of parameters including diffusion coefficients, binding constants, and photophysical rates, the framework describes fairly well the dynamic behavior of extra-synaptic and synaptic NLGN1 over both short and long time ranges, and provides an estimate of NLGN1 copy numbers in post-synaptic densities at steady-state (around 50 dimers). One striking result is that the residence time of NLGN1 at synapses is much longer than what can be expected from extracellular interactions with pre-synaptic neurexins only, suggesting that NLGN1 is stabilized at synapses through multivalent interactions with intracellular post-synaptic scaffolding proteins.

高分辨率荧光成像结合计算机模拟定量突触表面动力学和神经胶质素-1的纳米级组织
神经素(nlgn)形成了一个涉及突触发育的细胞粘附分子家族,但将这些蛋白保留在突触中的机制仍然不完全清楚。最近的研究表明,表面相关的NLGN1被弥漫性地困在突触中,在那里它与突触后密度的准静态支架元件相互作用。单分子追踪显示NLGN1在突触上的快速扩散和短暂固定在几秒钟内,而光漂白实验后的荧光恢复表明NLGN1在突触上的长期更新,在小时时间范围内。为了深入了解支持NLGN1在突触后锚定的机制,并试图调和这些实验范式,我们在这里定量分析了NLGN1的活细胞和超分辨率成像实验,使用新发布的荧光显微镜膜蛋白动力学模拟器FluoSim。基于一小部分参数,包括扩散系数、结合常数和光物理速率,该框架相当好地描述了突触外和突触内NLGN1在短时间和长时间范围内的动态行为,并提供了稳态(约50个二聚体)下突触后密度下NLGN1拷贝数的估计。一个引人注目的结果是NLGN1在突触中的停留时间比仅与突触前神经素的细胞外相互作用所能预期的要长得多,这表明NLGN1通过与细胞内突触后支架蛋白的多价相互作用在突触中稳定下来。
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
ACS Applied Bio Materials
ACS Applied Bio Materials Chemistry-Chemistry (all)
CiteScore
9.40
自引率
2.10%
发文量
464
期刊介绍: ACS Applied Bio Materials is an interdisciplinary journal publishing original research covering all aspects of biomaterials and biointerfaces including and beyond the traditional biosensing, biomedical and therapeutic applications. The journal is devoted to reports of new and original experimental and theoretical research of an applied nature that integrates knowledge in the areas of materials, engineering, physics, bioscience, and chemistry into important bio applications. The journal is specifically interested in work that addresses the relationship between structure and function and assesses the stability and degradation of materials under relevant environmental and biological conditions.
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