通过冷冻电镜对人工突触的超微结构特征进行无标记可视化。
Label-free visualization of ultrastructural features of artificial synapses via cryo-EM.
机构信息
Department of Chemistry, McGill University, 801 Sherbrooke Street West, H3A 2K6 Montreal, Canada; Montreal Neurological Institute, McGill University, 3801 University Street, H3A 2B4 Montreal, Canada.
出版信息
ACS Chem Neurosci. 2011 Dec 21;2(12):700-4. doi: 10.1021/cn200094j. Epub 2011 Oct 4.
Abstract
The ultrastructural details of presynapses formed between artificial substrates of submicrometer silica beads and hippocampal neurons are visualized via cryo-electron microscopy (cryo-EM). The silica beads are derivatized by poly-d-lysine or lipid bilayers. Molecular features known to exist at presynapses are clearly present at these artificial synapses, as visualized by cryo-EM. Key synaptic features such as the membrane contact area at synaptic junctions, the presynaptic bouton containing presynaptic vesicles, as well as microtubular structures can be identified. This is the first report of the direct, label-free observation of ultrastructural details of artificial synapses.
摘要
通过冷冻电子显微镜（cryo-EM）可视化了在亚微米二氧化硅珠和海马神经元之间形成的人工基质的突触前的超微结构细节。通过聚赖氨酸或脂质双层对二氧化硅珠进行衍生化。通过 cryo-EM 可以清楚地看到存在于这些人工突触中的已知存在于突触前的分子特征。可以识别出关键的突触特征，如突触连接处的膜接触面积、含有突触小泡的突触前小球，以及微管结构。这是直接、无标记观察人工突触超微结构细节的首次报道。
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本文引用的文献
1
Lipid bilayer membrane-triggered presynaptic vesicle assembly.脂质双层膜触发的突触前囊泡组装。
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2
Isolation of functional presynaptic complexes from CNS neurons: a cell-free preparation for the study of presynaptic compartments In vitro.从中枢神经系统神经元中分离功能性突触前复合物：体外研究突触前区室的无细胞制剂。
ACS Chem Neurosci. 2010 Aug 18;1(8):535-41. doi: 10.1021/cn100048z. Epub 2010 Jun 24.
3
Human MIEF1 recruits Drp1 to mitochondrial outer membranes and promotes mitochondrial fusion rather than fission.人源 MIEF1 募集 Drp1 至线粒体外膜并促进线粒体融合而非分裂。
EMBO J. 2011 Jun 24;30(14):2762-78. doi: 10.1038/emboj.2011.198.
4
Coordinated trafficking of synaptic vesicle and active zone proteins prior to synapse formation.突触前形成之前突触囊泡和活性区蛋白的协调转运。
Neural Dev. 2011 May 10;6:24. doi: 10.1186/1749-8104-6-24.
5
Mutant huntingtin binds the mitochondrial fission GTPase dynamin-related protein-1 and increases its enzymatic activity.突变型亨廷顿蛋白与线粒体分裂 GTP 酶相关蛋白 1 结合并增加其酶活性。
Nat Med. 2011 Mar;17(3):377-82. doi: 10.1038/nm.2313. Epub 2011 Feb 20.
6
Quantitative analysis of the native presynaptic cytomatrix by cryoelectron tomography.通过冷冻电镜断层扫描对天然突触前细胞外基质进行定量分析。
J Cell Biol. 2010 Jan 11;188(1):145-56. doi: 10.1083/jcb.200908082.
7
Rapid assembly of functional presynaptic boutons triggered by adhesive contacts.由黏附接触触发的功能性突触前终扣的快速组装。
J Neurosci. 2009 Oct 7;29(40):12449-66. doi: 10.1523/JNEUROSCI.1381-09.2009.
8
Supported bilayers formed from different phospholipids on spherical silica substrates.由不同磷脂在球形二氧化硅基底上形成的支撑双层膜。
Langmuir. 2009 May 19;25(10):5455-8. doi: 10.1021/la9006982.
9
Synaptic transmission of neural stem cells seeded in 3-dimensional PLGA scaffolds.接种于三维聚乳酸-羟基乙酸共聚物支架中的神经干细胞的突触传递。
Biomaterials. 2009 Aug;30(22):3711-22. doi: 10.1016/j.biomaterials.2009.03.046. Epub 2009 Apr 17.
10
Resolving sub-synaptic compartments with double immunofluorescence labeling in hippocampal neurons.利用双重免疫荧光标记解析海马神经元中的亚突触区室
J Neurosci Methods. 2009 Jan 30;176(2):78-84. doi: 10.1016/j.jneumeth.2008.08.025. Epub 2008 Aug 30.
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