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观察原代小鼠神经元和小胶质细胞固定及活细胞共培养体系中隧道纳米管形成及功能的实验方案。

Protocol for observing tunneling nanotube formation and function in both fixed and live primary mouse neurons and microglia coculture system.

作者信息

Baker Vivian, Budinger Dimitri, Riechers Sean-Patrick, Heneka Michael T

机构信息

Luxembourg Centre for Systems Biomedicine, University of Luxembourg, 6 Avenue du Swing, 4367 Esch-Belval, Luxembourg.

Luxembourg Centre for Systems Biomedicine, University of Luxembourg, 6 Avenue du Swing, 4367 Esch-Belval, Luxembourg.

出版信息

STAR Protoc. 2025 Apr 3;6(2):103723. doi: 10.1016/j.xpro.2025.103723.

DOI:10.1016/j.xpro.2025.103723
PMID:40184246
原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC12002979/
Abstract

Microglia and neurons can connect via tunneling nanotubes (TNTs), facilitating the transfer of organelles, vesicles, and proteins. Here, we present a protocol for visualizing murine TNT formation and material transfer between neurons and microglia in both fixed samples and samples for live-cell imaging, as well as for flow cytometry. We describe steps for identifying and measuring TNTs and quantifying the transport of aggregated proteins, such as α-synuclein or tau, between these cells. For complete details on the use and execution of this protocol, please refer to Scheiblich et al..

摘要

小胶质细胞和神经元可通过隧道纳米管（TNTs）相连，促进细胞器、囊泡和蛋白质的转移。在此，我们提供了一种方法，用于在固定样本以及活细胞成像样本和流式细胞术样本中观察小鼠TNTs的形成以及神经元与小胶质细胞之间的物质转移。我们描述了识别和测量TNTs以及量化这些细胞之间聚集蛋白（如α-突触核蛋白或tau蛋白）转运的步骤。有关该方法的使用和执行的完整详细信息，请参考Scheiblich等人的研究。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/dba7/12002979/bcad7d2220dd/gr8.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/dba7/12002979/8a27b9b4c154/fx1.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/dba7/12002979/74c7687ae376/gr1.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/dba7/12002979/eba66e11cfe0/gr2.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/dba7/12002979/a7a0791edb62/gr3.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/dba7/12002979/cb514e9f3aee/gr4.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/dba7/12002979/1d0c17f17410/gr5.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/dba7/12002979/6f16509d335d/gr6.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/dba7/12002979/f30d30244d8a/gr7.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/dba7/12002979/bcad7d2220dd/gr8.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/dba7/12002979/8a27b9b4c154/fx1.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/dba7/12002979/74c7687ae376/gr1.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/dba7/12002979/eba66e11cfe0/gr2.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/dba7/12002979/a7a0791edb62/gr3.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/dba7/12002979/cb514e9f3aee/gr4.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/dba7/12002979/1d0c17f17410/gr5.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/dba7/12002979/6f16509d335d/gr6.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/dba7/12002979/f30d30244d8a/gr7.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/dba7/12002979/bcad7d2220dd/gr8.jpg

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Microglia rescue neurons from aggregate-induced neuronal dysfunction and death through tunneling nanotubes.小胶质细胞通过形成隧道纳米管来挽救聚集诱导的神经元功能障碍和死亡。
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