Name: 3d modeling of dendritic spines with synaptic plasticity
Uploaded: 2020-05-18T13:30:00
Description: Watch this Scientific Journal Video about 3D Modeling of Dendritic Spines with Synaptic Plasticity at JoVE.com

这项工作得到了圣保罗州科学基金会#2015/50122-0和IRTG-GRTK 1740/2赠款的一部分支持，IBM/FAPESP赠款#2016/18825-4和FAPESP赠款#2018/06504-4。

注：有关本协议中使用的术语表，请参阅补充文件1。
1. 安装搅拌机、电池搅拌机和 MCell
注：此协议需要安装 MCell、搅拌机和细胞搅拌机。
<ol><li>下载并安装 MCell 主页（https://mcell.org/tutorials_iframe.html）。转到页面顶部的下载，然后按照分步说明下载并在首选环境中（例如 Linux、Mac OSX 或 Windows）安装软件。 注：本协议中描述的所有计算模?...

<ol>
	<li>Sweatt, J. D. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Neural+plasticity+and+behavior+-+sixty+years+of+conceptual+advances.">Neural plasticity and behavior - sixty years of conceptual advances.</a> Journal of Neurochemistry. 139, 179-199 (2016).</li><li>Heine, M., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Surface+mobility+of+postsynaptic+AMPARs+tunes+synaptic+transmission.">Surface mobility of postsynaptic AMPARs tunes synaptic transmission.</a> Science. 320 (5873), 201-205 (2008).</li><li>Buonarati, O. R., Hammes, E. A., Watson, J. F., Greger, I. H., Hell, J. W. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Mechanisms+of+postsynaptic+localization+of+AMPA-type+glutamate+receptors+and+their+regulation+during+long-term+potentiation.">Mechanisms of postsynaptic localization of AMPA-type glutamate receptors and their regulation during long-term potentiation.</a> Science Signaling. 12 (562), 6889 (2019).</li><li>Nair, D., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Super-Resolution+Imaging+Reveals+That+AMPA+Receptors+Inside+Synapses+Are+Dynamically+Organized+in+Nanodomains+Regulated+by+PSD95.">Super-Resolution Imaging Reveals That AMPA Receptors Inside Synapses Are Dynamically Organized in Nanodomains Regulated by PSD95.</a> Journal of Neuroscience. 33 (32), 13204-13224 (2013).</li><li>Cz&#246;nd&#246;r, K., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Unified+quantitative+model+of+AMPA+receptor+trafficking+at+synapses.">Unified quantitative model of AMPA receptor trafficking at synapses.</a> Proceeding of the National Academy of Sciences of the United States of America. 109 (9), 3522-3527 (2012).</li><li>Triesch, J., Vo, A. D., Hafner, A. S. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Competition+for+synaptic+building+blocks+shapes+synaptic+plasticity.">Competition for synaptic building blocks shapes synaptic plasticity.</a> eLife. 7, 37836 (2018).</li><li>Earnshaw, B. A., Bressloff, P. C. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Biophysical+model+of+AMPA+receptor+trafficking+and+its+regulation+during+long-term+potentiation/long-term+depression.">Biophysical model of AMPA receptor trafficking and its regulation during long-term potentiation/long-term depression.</a> Journal of Neuroscience. 26 (47), 12362-12373 (2006).</li><li>Earnshaw, B. A., Bressloff, P. C. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Modeling+the+role+of+lateral+membrane+diffusion+in+AMPA+receptor+trafficking+along+a+spiny+dendrite.">Modeling the role of lateral membrane diffusion in AMPA receptor trafficking along a spiny dendrite.</a> Journal of Computational Neuroscience. 25 (2), 366-389 (2008).</li><li>Antunes, G., Roque, A. C., Simoes-de-Souza, F. M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Stochastic+Induction+of+Long-Term+Potentiation+and+Long-Term+Depression.">Stochastic Induction of Long-Term Potentiation and Long-Term Depression.</a> Scientific Reports. 6, 30899 (2016).</li><li>Kotaleski, J. H., Blackwell, K. T. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Modelling+the+molecular+mechanisms+of+synaptic+plasticity+using+systems+biology+approaches.">Modelling the molecular mechanisms of synaptic plasticity using systems biology approaches.</a> Nature Reviews Neuroscience. 11 (4), 239-251 (2010).</li><li>Bhalla, U. S. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Molecular+computation+in+neurons:+a+modeling+perspective.">Molecular computation in neurons: a modeling perspective.</a> Current Opinion in Neurobiology. 25, 31-37 (2014).</li><li>Cz&#246;nd&#246;r, K., Thoumine, O. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Biophysical+mechanisms+regulating+AMPA+receptor+accumulation+at+synapses.">Biophysical mechanisms regulating AMPA receptor accumulation at synapses.</a> Brain Research Bulletin. 93, 57-68 (2013).</li><li>Bromer, C., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Long-term+potentiation+expands+information+content+of+hippocampal+dentate+gyrus+synapses.">Long-term potentiation expands information content of hippocampal dentate gyrus synapses.</a> Proceedings of the National Academy of Sciences of the United States of America. 115 (10), 2410-2418 (2018).</li><li>Antunes, G., Simoes-de-Souza, F. M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=AMPA+receptor+trafficking+and+its+role+in+heterosynaptic+plasticity.">AMPA receptor trafficking and its role in heterosynaptic plasticity.</a> Scientific Reports. 8 (1), 10349 (2018).</li><li>Kerr, R. A., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Fast+monte+carlo+simulation+methods+for+biological+reaction-diffusion+systems+in+solution+and+on+surfaces.">Fast monte carlo simulation methods for biological reaction-diffusion systems in solution and on surfaces.</a> SIAM Journal on Scientific Computing. 30 (6), 3126 (2008).</li><li>Czech, J., Dittrich, M., Stiles, J. R. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Rapid+Creation,+Monte+Carlo+Simulation,+and+Visualization+of+Realistic+3D+Cell+Models.">Rapid Creation, Monte Carlo Simulation, and Visualization of Realistic 3D Cell Models.</a> Systems Biology. 500, 237-287 (2009).</li><li>Stiles, J., Bartol, T., De Schutter, <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Monte+Carlo+Methods+for+Simulating+Realistic+Synaptic+Microphysiology+Using+MCell.">Monte Carlo Methods for Simulating Realistic Synaptic Microphysiology Using MCell.</a> Computational Neuroscience. , (2000).</li><li>Jorstad, A., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=NeuroMorph:+A+Toolset+for+the+Morphometric+Analysis+and+Visualization+of+3D+Models+Derived+from+Electron+Microscopy+Image+Stacks.">NeuroMorph: A Toolset for the Morphometric Analysis and Visualization of 3D Models Derived from Electron Microscopy Image Stacks.</a> Neuroinformatics. 13 (1), 83-92 (2015).</li><li>Antunes, G., Roque, A. C., Simoes de Souza, F. M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Modelling+intracellular+competition+for+calcium:+kinetic+and+thermodynamic+control+of+different+molecular+modes+of+signal+decoding.">Modelling intracellular competition for calcium: kinetic and thermodynamic control of different molecular modes of signal decoding.</a> Scientific Reports. 6, 23730 (2016).</li><li>Antunes, G., Roque, A. C., Simoes-de-Souza, F. M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Molecular+mechanisms+of+detection+and+discrimination+of+dynamic+signals.">Molecular mechanisms of detection and discrimination of dynamic signals.</a> Scientific Reports. 8 (1), 2480 (2018).</li><li>Hoops, S., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=COPASI--a+COmplex+PAthway+SImulator.">COPASI--a COmplex PAthway SImulator.</a> Bioinformatics. 22 (24), 3067-3074 (2006).</li><li>Faeder, J. R., Blinov, M. L., Hlavacek, W. S. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Rule-based+modeling+of+biochemical+systems+with+BioNetGen.">Rule-based modeling of biochemical systems with BioNetGen.</a> Methods in Molecular Biology. 500, 113-167 (2009).</li><li>Gillespie, D. T. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Exact+stochastic+simulation+of+coupled+chemical+reactions.">Exact stochastic simulation of coupled chemical reactions.</a> Journal of Physical Chemistry. 81 (25), 21 (1977).</li><li>Anggono, V., Huganir, R. L. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Regulation+of+AMPA+receptor+trafficking+and+synaptic+plasticity.">Regulation of AMPA receptor trafficking and synaptic plasticity.</a> Current Opinion in Neurobiology. 22 (3), 461-469 (2012).</li><li>Matsuda, S., Launey, T., Mikawa, S., Hirai, H. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Disruption+of+AMPA+receptor+GluR2+clusters+following+long-term+depression+induction+in+cerebellar+Purkinje+neurons.">Disruption of AMPA receptor GluR2 clusters following long-term depression induction in cerebellar Purkinje neurons.</a> EMBO Journal. 19 (12), 2765-2774 (2000).</li><li>Ahmad, M., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Postsynaptic+Complexin+Controls+AMPA+Receptor+Exocytosis+during+LTP.">Postsynaptic Complexin Controls AMPA Receptor Exocytosis during LTP.</a> Neuron. 73 (2), 260-267 (2012).</li><li>Sheng, M., Hoogenraad, C. C. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=The+postsynaptic+architecture+of+excitatory+synapses:+a+more+quantitative+view.">The postsynaptic architecture of excitatory synapses: a more quantitative view.</a> Annual Review of Biochemistry. 76, 823-847 (2007).</li></ol>

本文提出了一种构建3D网格的方法，用于在树突状脊柱的树突状段中对反应-扩散突触可塑性过程进行建模。开发的模型包含一个树突状段，很少树突状脊柱。带突触锚的 AMPAR 的横向扩散和反应允许模拟基础动力学。协议中的关键步骤是切割球体以创建脊柱头的顶部（图1，图2，图3），创建脊柱颈部的挤出（图4）?...

突触可塑性与学习和记忆1相关。突触可塑性，如长期增效（LTP）和长期抑郁症（LTD），分别与突触膜2中和外突触膜中插入和去除AMPA受体（AMPAR）相关。AMPAR突触位于称为树突状脊柱3的小体积结构的顶部。每个脊柱在后突触膜中含有一个蛋白质密集区域，称为后突触密度（PSD）。在PSD捕获的突触区域的安培蛋白。单个突触中很少有AMPAR的拷贝，AMPAR与树突状脊柱中其他物种的贩运和反应是一个随机过程2,2，4。在树突状脊柱5，6，7，8，突,触受体贩运有几个7分门别类模型。然而，在树突及其树突状脊柱的3D结构中，缺乏与突触可塑性相关的AMPAR的贩运的随机计算模型。
计算建模是研究复杂系统动力学基础的有用工具，如突触可塑性,9、10、11、12期间突,10触可塑性在树突脊柱中的反应11,扩散。该模型可用于可视化复杂的场景，改变敏感参数，并在科学条件下进行重要的预测，涉及许多难以或不可能控制实验12、13,的变量。定义计算模型的细节级别是获取有关建模现象的准确信息的基本步骤。理想的计算模型是复杂性和简单性之间的微妙平衡，以捕获自然现象的基本特征，而不会在计算上令人望而却步。过于详细的计算模型的计算成本可能很高。另一方面，细节不详的系统可能缺乏捕捉这一现象动态所必须的基本组成部分。尽管树突脊柱的 3D 建模在计算上比 2D 和 1D 更昂贵，但存在一些条件，例如在具有许多非线性变量在时间和 3D 空间中发生反应和扩散的复杂系统中，对于 3D 级别的建模对于获得有关系统功能的见解至关重要。此外，可以仔细降低复杂性，以保持低维模型的基本特征。
在小体积内少量复制给定物种的随机系统中，系统的平均动力学与大量种群的平均动力学不同。在这种情况下，需要反应扩散粒子的随机计算建模。本文介绍了一种在3D树突脊柱中对几份AMPAR进行随机建模的方法。此方法的目的是开发一个树突状段的三维计算模型与树突状脊柱及其突触建模突触可塑性。
该方法使用软件MCell求解模型，用于构建3D网格的Blender，使用CellBlender创建和可视化MCell模拟，包括3D网格14、15、16,15中的分子的时空反应扩散16。搅拌机是用于创建网格的套件，而 CellBlender 是基本软件 Blender 的附加设备。MCell是一个蒙特卡罗模拟器，用于单个分子17的反应扩散。
使用这种方法的原理包括对突触可塑性进行建模，以便更好地了解树突状脊柱14的微观生理环境中的这一现象。特别是，这种方法允许模拟同突电位化，同突增抑郁症，和异质突触可塑性之间的树突状脊柱14。
该方法的特点包括树突及其突触的三维几何结构建模，随机行走扩散，以及突触可塑性分子的化学反应。此方法提供了创建丰富的环境以测试假设和预测具有大量变量的复杂非线性系统功能的优势。此外，该方法不仅可用于研究突触可塑性，还可用于研究三维网格结构中分子的随机反应扩散。
或者，树突结构的3D网格可以直接在搅拌机中从电子显微镜串行重建18中构建。尽管基于串行重建的网格提供 3D 结构，但访问实验数据并非始终可用。因此，如本协议所述，根据基本几何结构改编的网格结构提供了灵活性，可以开发具有树突状脊柱的定制树突状段。
另一种替代计算方法是在,常规卷,,9、10、11、19、20、21、22中对混合反应21进行批量模拟。20,10,11919散装模拟在解决单个混合体积23中许多物种的反应方面非常有效，但散装方法在高分辨率3D网格中解决许多混合良好的体素中分子的反应扩散速度非常慢。另一方面，目前采用MCell模拟单个粒子反应扩散的方法在高分辨率3D网格15中有效工作。
在使用此方法之前，应询问所研究的现象是否需要在 3D 网格中采用随机反应扩散方法。如果该现象的至少一个反应物种的拷贝（少于 1，000 个）在复杂的几何结构中扩散，具有小体积隔间（如树突状脊柱），则 3D 网格中反应扩散的随机建模适合应用。
构建包含突触可塑性树突状脊柱的树突状段的 3D 计算模型需要几个步骤。主要步骤是安装用于构建模型的适当软件，构建一个用作创建多个脊柱的模板的单个树突脊柱，以及创建与多个树突脊柱相连的树突段。模拟突触可塑性的步骤包括插入 PSD 区域的锚点和树突状段和树突状脊柱中的 AMPAR。然后，在位于PSD和AMPAR的锚点之间的动力学反应被定义产生复杂的锚-AMPAR物种，在突触区域捕获AMPAR。分别，锚点和突触安培之间的亲和力的增加和减少创造了LTP和LTC的过程。

<table>
	<tbody>
		<tr>
			<td>Blender</td>
			<td>Blender Foundation</td>
			<td></td>
			<td>https://www.blender.org/</td>
		</tr>
		<tr>
			<td>CellBlender</td>
			<td>University of Pittsburgh</td>
			<td></td>
			<td>https://mcell.org/</td>
		</tr>
		<tr>
			<td>Mcell</td>
			<td>University of Pittsburgh</td>
			<td></td>
			<td>https://mcell.org/</td>
		</tr>
	</tbody>
</table>

3d modeling of dendritic spines with synaptic plasticity

三维（3D）几何体中化学物种的扩散和反应的计算建模是理解突触脊柱突触可塑性机制的基本方法。在此协议中，树突和树突脊柱的详细 3D 结构使用软件搅拌机与 CellBlender 上的网格进行建模。突触和突触外突触区域在网格上定义。接下来，突触受体和突触锚分子用其扩散常数定义。最后，纳入了突触受体与突触锚之间的化学反应，并采用软件MCell对计算模型进行数值解。该方法描述了三维几何结构中每个分子的时空路径。因此，在突触可塑性发生期间，研究突触受体在树突状脊柱的贩运是非常有用的。这种方法的一个局限性是，大量的分子会减慢模拟的速度。使用这种方法对树突状脊柱进行建模，可以研究单脊柱内的同突突起和抑郁，以及相邻树突状脊柱之间的异质突增。

这些结果提供了构建 3D 网格的步骤，该网格模拟具有脊柱头和脊柱颈部的树突状脊柱（图 1 到图 4）。此外，多个树突状脊柱可以插入单个树突状段（图5），以研究AMPARs14的异质突增性。脊柱头顶部的 PSD（图 6）是突触锚点与 AMPAR 结合并暂时捕获突触的位置（图 7，<...

Watch this Scientific Journal Video about 3D Modeling of Dendritic Spines with Synaptic Plasticity at JoVE.com

3D Modeling of Dendritic Spines with Synaptic Plasticity

该协议开发一个三维（3D）模型的树突状段与树突状脊柱建模突触可塑性。构造网格可用于计算建模的AMPA受体贩运在长期突触可塑性使用的软件程序搅拌机与细胞Blender和MCell。

具有突触可塑性树突脊柱的三维建模

Computational modeling of diffusion and reaction of chemical species in a three-dimensional (3D) geometry is a fundamental method to understand the ...

通过融合对化学物种反应的计算三维几何建模。是研究树突脊柱内和外树突的受体贩运机制的有用方法，是做突触可塑性。此技术的优点是创建丰富的环境，用于对具有大量变量的非线性系统进行假设和预测。使用修改后的球体创建具有脊柱头和脊柱颈部的单个树突脊柱的网格。第一次打开搅拌机。在键盘上已安装 Cell Blender 并按 5 键，从透视更改为正交视图。按 1 更改为前视图，按 Shift C 以居中光标。要创建脊柱头部，请按 Shift A 打开"网格"调色板，然后选择"网格"。选择 UV 球体并在添加 UV 球体中，将大小设置为 0.25，将环设置为 32。要使头部顶部平坦，请按 Tab 将搅拌机从对象模式切换到"编辑"模式。按 B 选择球体的前三个季度，然后按"删除"，选择"顶点"和"输入"以删除顶点。按 B 并选择球体的顶部。按 E、S、0 和 Enter 以密封仍所选的顶点的顶部。向下移动蓝色箭头以对齐脊柱头的顶部。要增加脊柱顶部的网格分辨率，请选择"工具和刀"，然后使用刀在顶部中心周围切割一个圆圈。然后选择"工具和循环切割"并滑动四次，以围绕顶部中心创建四个同心圆。要创建脊柱颈部，请按 B 并选择网格的底部。按"删除顶点"和"B"并选择网格的底部。按 E 和 Z 并选择负 0.45 以在负 0.45 微米时创建到 Z 轴位置的拉伸。要使网格与 M 单元格兼容，请按 Ctrl T 对网格进行三角测量，然后选择"工具"和"删除双倍"。要创建多个脊椎 Dendrite，请按 Shift A 打开网格调色板并选择"网格"和"圆柱"。在"添加圆柱体"菜单中，将半径 0.3 微米和深度设置为两微米，然后按 Enter。 按 R 并输入 90 以旋转圆柱体 90 度，然后使用蓝色箭头将圆柱体拖动到脊柱底部。按 3 可获取圆柱体的前视图，然后按 Z 使网格透明。使用蓝色法线箭头将脊柱底部移动到圆柱体内部，右键单击以选择 Dendrite。选择修改器和添加修改器，然后选择"布尔"，"操作联合"，然后选择对象脊柱。单击"应用"以创建登狄石和脊柱的关节网格。然后使用鼠标选择隔离脊柱的网格，更改位置和角度以将每个新脊柱插入生理位置。要创建 AMPAR，请选择分子并插入新分子。将名称更改为 AMPAR，将分子类型更改为表面分子。然后将扩散常数 0.05 倍 10 更改为 8 平方厘米/秒。要绘制与 AMPAR 绑定的锚点，在基础条件期间在 PSD1 下，请打开"绘图输出设置"并按以定义分子。然后将分子设置为Anchor_AMPAR，将对象设置为登狄石，将区域设置为 PSD1。要运行基础条件模拟，请选择"运行模拟"。将迭代设置为 30，000，将时间步长设置为 1 倍 10 到负 3 秒。单击"导出并运行"并等待模拟结束。在模拟结束时，选择"重新加载可视化数据"、播放动画、绘制输出设置和绘图以可视化空间时间结果。要运行同性突触电位条件，选择分子放置rel_anchorLTP_psd1。选择rel_anchorLTP_psd1，将"数量"更改为"200"。然后将"数量"更改为"释放"为"零"。选择rel_anchor_psd1。将数量更改为零，并运行模拟，正如刚才演示的。突触可塑性可以通过每个脊柱锚点固定数量的 AMPAR 数量的变化大致验证。用于精确计算突触可塑性的发生。建议计算突触上锚定和自由安培的总数的变化。AMPAR同性突触和凹陷可以通过分别增加和减少锚定安培的数量来验证，这些安培的亲和力与基底条件相比，由锚点引起的亲和力发生变化。例如，单一脊柱的同性突触长期电能诱导，在相邻脊柱上产生异质突触长期抑郁效应。按照此过程，可以扩展模型，以调查登得利脊柱的 LTP 和 LTD 感应过程。此方法允许测试有关具有大量变量的复杂非线性系统功能的假设。

Research

JoVE Journal

Neuroscience

Preparation of Oligomeric &beta;-amyloid1-42 and Induction of Synaptic Plasticity Impairment on Hippocampal Slices

Preparation of Oligomeric ÃƒÅ½Ã‚Â²-amyloid1-42 and Induction of Synaptic Plasticity Impairment on Hippocampal Slices

低聚β-淀粉样蛋白的制备 1-42和突触可塑性损伤诱导海马脑片

Impairment of synaptic connections is likely to underlie the subtle amnesic changes occurring at the early stages of Alzheimer s Disease (AD). ...

科研

神经科学

Fluorescence Recovery After Photobleaching (FRAP) of Fluorescence Tagged Proteins in Dendritic Spines of Cultured Hippocampal Neurons

Fluorescence Recovery After Photobleaching FRAP of Fluorescence Tagged Proteins in Dendritic Spines of Cultured Hippocampal Neurons

漂白后荧光标记蛋白的荧光恢复（FRAP）在培养海马神经元的树突棘

FRAP has been used to quantify the mobility of GFP-tagged proteins. Using a strong excitation laser, the fluorescence of a GFP-tagged protein is bleached ...

Lateral Diffusion and Exocytosis of Membrane Proteins in Cultured Neurons Assessed using Fluorescence Recovery and Fluorescence-loss Photobleaching

横向扩散和培养的神经元膜蛋白的胞吐评估使用荧光恢复和荧光损失漂白

Membrane proteins such as receptors and ion channels undergo active trafficking in neurons, which are highly polarised and morphologically complex. This ...

Voltage-sensitive Dye Recording from Axons, Dendrites and Dendritic Spines of Individual Neurons in Brain Slices

电压敏感染料记录单个神经元在大脑切片的轴突，树突和树突棘

Understanding the biophysical properties and functional organization of single neurons and how they process information is fundamental for understanding ...

Fast Micro-iontophoresis of Glutamate and GABA: A Useful Tool to Investigate Synaptic Integration

谷氨酸和GABA快速微离子导入：一个有用​​的工具，调查突触整合

One of the fundamental interests in neuroscience is to understand the integration of excitatory and inhibitory inputs along the very complex structure of ...

Deriving the Time Course of Glutamate Clearance with a Deconvolution Analysis of Astrocytic Transporter Currents

星形细胞转运电流派生谷氨酸间隙的时间进程与反卷积分析

The highest density of glutamate transporters in the brain is found in astrocytes. Glutamate transporters couple the movement of glutamate across the ...

Imaging Dendritic Spines of Rat Primary Hippocampal Neurons using Structured Illumination Microscopy

成像大鼠原代海马神经元树突棘使用结构照明显微镜

Dendritic spines are protrusions emerging from the dendrite of a neuron and represent the primary postsynaptic targets of excitatory inputs in the brain. ...

Two-Photon in vivo Imaging of Dendritic Spines in the Mouse Cortex Using a Thinned-skull Preparation

Two-Photon in vivo Imaging of Dendritic Spines in the Mouse Cortex Using a Thinned-skull Preparation

双光子<em&gt;在体内</em&gt;树突棘中的鼠标皮质用稀疏的头颅成像准备

In the mammalian cortex, neurons form extremely complicated networks and exchange information at synapses. Changes in synaptic strength, as well as ...

Pentylenetetrazole-Induced Kindling Mouse Model

戊四氮诱导点燃小鼠模型

Pentylenetetrazole (PTZ) is a GABA-A receptor antagonist. An intraperitoneal injection of PTZ into an animal induces an acute, severe seizure at a high ...

Combining Optogenetics with Artificial microRNAs to Characterize the Effects of Gene Knockdown on Presynaptic Function within Intact Neuronal Circuits

光遗传学与人工 microRNAs 相结合对完整神经元回路中基因击倒对突触前功能的影响

The purpose of this protocol is to characterize the effect of gene knockdown on presynaptic function within intact neuronal circuits. We describe a ...