神经肌肉接头处的FM1-43光转化与电子显微镜分析
FM1-43 Photoconversion and Electron Microscopy Analysis at the Neuromuscular Junction.
作者信息
Sabeva Nadezhda S, Bykhovskaia Maria
机构信息
Neuroscience Department, Universidad Central del Caribe, Bayamón, Puerto Rico, USA.
Department of Neurology, Department of Anatomy and Cell Biology, Wayne State University School of Medicine, Detroit, Michigan, USA.
出版信息
Bio Protoc. 2017 Sep 5;7(17). doi: 10.21769/BioProtoc.2523.
Abstract
We developed a protocol for photoconversion of endocytic marker FM1-43 followed by electron microscopy analysis of synaptic boutons at the neuromuscular junction. This protocol allows detection of stained synaptic vesicle even when release rates are very low, such as during the spontaneous release mode. The preparations are loaded with the FM1-43 dye, pre-fixed, treated and illuminated to photoconvert the dye, and then processed for conventional electron microscopy. This procedure enables clear identification of stained synaptic vesicles at electron micrographs.
摘要
我们开发了一种用于内吞标记物FM1-43光转化的方案，随后对神经肌肉接头处的突触小体进行电子显微镜分析。即使在释放速率非常低的情况下，如在自发释放模式期间，该方案也能检测到染色的突触小泡。将制剂用FM1-43染料加载、预固定、处理并照射以光转化染料，然后进行常规电子显微镜处理。该程序能够在电子显微镜照片上清晰地识别染色的突触小泡。
相似文献
1
FM1-43 Photoconversion and Electron Microscopy Analysis at the Neuromuscular Junction.神经肌肉接头处的FM1-43光转化与电子显微镜分析
Bio Protoc. 2017 Sep 5;7(17). doi: 10.21769/BioProtoc.2523.
2
Complexin Mutants Reveal Partial Segregation between Recycling Pathways That Drive Evoked and Spontaneous Neurotransmission.复合体蛋白突变体揭示了驱动诱发和自发神经传递的回收途径之间的部分分离。
J Neurosci. 2017 Jan 11;37(2):383-396. doi: 10.1523/JNEUROSCI.1854-16.2016.
3
Synapsin regulates vesicle organization and activity-dependent recycling at Drosophila motor boutons.突触结合蛋白调节果蝇运动终板处囊泡的组织和活动依赖性循环。
Neuroscience. 2010 Oct 13;170(2):441-52. doi: 10.1016/j.neuroscience.2010.07.021. Epub 2010 Jul 16.
4
Synapsin maintains the reserve vesicle pool and spatial segregation of the recycling pool in Drosophila presynaptic boutons.突触结合蛋白维持果蝇突触前终扣中储备囊泡池和循环池的空间隔离。
Brain Res. 2007 Oct 31;1178:52-64. doi: 10.1016/j.brainres.2007.08.042. Epub 2007 Aug 24.
5
Studying synaptic vesicle pools using photoconversion of styryl dyes.利用苯乙烯基染料的光转化研究突触小泡池。
J Vis Exp. 2010 Feb 15(36):1790. doi: 10.3791/1790.
6
Stimulation-induced formation of the reserve pool of vesicles in Drosophila motor boutons.刺激诱导果蝇运动终扣中囊泡储备池的形成。
J Neurophysiol. 2009 May;101(5):2423-33. doi: 10.1152/jn.91122.2008. Epub 2009 Mar 11.
7
Time and exposure to serotonin affect releasability of recycled vesicles at crayfish claw opener muscle synapses.时间和暴露于血清素会影响螯虾爪开肌突触中再循环囊泡的可释放性。
Synapse. 2020 Feb;74(2):e22136. doi: 10.1002/syn.22136. Epub 2019 Nov 10.
8
Membrane cholesterol regulates different modes of synaptic vesicle release and retrieval at the frog neuromuscular junction.膜胆固醇调节青蛙神经肌肉接头处突触小泡释放和回收的不同模式。
Eur J Neurosci. 2013 Oct;38(7):2978-87. doi: 10.1111/ejn.12300. Epub 2013 Jul 10.
9
Exocytosis and endocytosis of synaptic vesicles and functional roles of vesicle pools: lessons from the Drosophila neuromuscular junction.突触小泡的胞吐作用与胞吞作用以及小泡池的功能作用：来自果蝇神经肌肉接头的启示
Neuroscientist. 2005 Apr;11(2):138-47. doi: 10.1177/1073858404271679.
10
FM1-43 dye ultrastructural localization in and release from frog motor nerve terminals.FM1-43染料在青蛙运动神经末梢中的超微结构定位及其从末梢的释放
Proc Natl Acad Sci U S A. 1996 Mar 5;93(5):1918-23. doi: 10.1073/pnas.93.5.1918.
引用本文的文献
1
Immuno-electrophysiology on Neuromuscular Junctions of Drosophila Third Instar Larva.果蝇三龄幼虫神经肌肉接头的免疫电生理学研究
Bio Protoc. 2021 Feb 5;11(3):e3913. doi: 10.21769/BioProtoc.3913.
2
Dedicated Setup for the Photoconversion of Fluorescent Dyes for Functional Electron Microscopy.用于功能电子显微镜荧光染料光转换的专用装置。
Front Cell Neurosci. 2019 Jul 24;13:312. doi: 10.3389/fncel.2019.00312. eCollection 2019.
3
Two Pathways for the Activity-Dependent Growth and Differentiation of Synaptic Boutons in .突触末梢活性依赖的生长和分化的两种途径。
eNeuro. 2019 Aug 22;6(4). doi: 10.1523/ENEURO.0060-19.2019. Print 2019 Jul/Aug.
4
Ptp4E regulates vesicular packaging for monoamine-neuropeptide co-transmission.Ptp4E 调节单胺-神经肽共传递的囊泡包装。
J Cell Sci. 2019 Apr 3;132(7):jcs224568. doi: 10.1242/jcs.224568.
5
FM Dye Cycling at the Synapse: Comparing High Potassium Depolarization, Electrical and Channelrhodopsin Stimulation.突触处的荧光染料循环：比较高钾去极化、电刺激和通道视紫红质刺激
J Vis Exp. 2018 May 24(135):57765. doi: 10.3791/57765.
本文引用的文献
1
Complexin Mutants Reveal Partial Segregation between Recycling Pathways That Drive Evoked and Spontaneous Neurotransmission.复合体蛋白突变体揭示了驱动诱发和自发神经传递的回收途径之间的部分分离。
J Neurosci. 2017 Jan 11;37(2):383-396. doi: 10.1523/JNEUROSCI.1854-16.2016.
2
Revisiting synaptic vesicle pool localization in the Drosophila neuromuscular junction.重新审视果蝇神经肌肉接头处突触小泡池的定位
J Physiol. 2009 Jun 15;587(Pt 12):2919-26. doi: 10.1113/jphysiol.2009.170985. Epub 2009 Apr 29.
3
Enhancement of the endosomal endocytic pathway increases quantal size.内体胞吞途径的增强会增加量子大小。
Mol Cell Neurosci. 2009 Feb;40(2):199-206. doi: 10.1016/j.mcn.2008.10.005. Epub 2008 Nov 5.
4
FM 1-43 labeling of synaptic vesicle pools at the Drosophila neuromuscular junction.果蝇神经肌肉接头处突触小泡池的FM 1-43标记
Methods Mol Biol. 2008;440:349-69. doi: 10.1007/978-1-59745-178-9_26.
5
A complexin fusion clamp regulates spontaneous neurotransmitter release and synaptic growth.一种复合体融合钳调控自发性神经递质释放和突触生长。
Nat Neurosci. 2007 Oct;10(10):1235-7. doi: 10.1038/nn1980. Epub 2007 Sep 16.
6
Synaptic vesicle reuse and its implications.突触小泡的再利用及其意义。
Neuroscientist. 2006 Feb;12(1):57-66. doi: 10.1177/1073858405281852.
7
Synaptic vesicle pools.突触小泡池
Nat Rev Neurosci. 2005 Jan;6(1):57-69. doi: 10.1038/nrn1583.
8
The synaptic vesicle cycle.突触小泡循环。
Annu Rev Neurosci. 2004;27:509-47. doi: 10.1146/annurev.neuro.26.041002.131412.
9
The structural organization of the readily releasable pool of synaptic vesicles.突触小泡易释放池的结构组织
Science. 2004 Mar 26;303(5666):2037-9. doi: 10.1126/science.1094682.
10
Visualizing recycling synaptic vesicles in hippocampal neurons by FM 1-43 photoconversion.通过FM 1-43光转化可视化海马神经元中循环的突触小泡。
Proc Natl Acad Sci U S A. 2001 Oct 23;98(22):12748-53. doi: 10.1073/pnas.171442798.
Zotero 插件