Name: recording and modulation of epileptiform activity in rodent brain slices coupled to microelectrode arrays
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Description: Watch this Scientific Journal Video about Recording and Modulation of Epileptiform Activity in Rodent Brain Slices Coupled to Microelectrode Arrays at JoVE.com

他是由欧洲联盟 MSCA-IF-2014 项目资助的玛丽居里夫人-居里研究员. b. 美国, g.a.n. 660689, 框架计划 H2020。GUI mapMEA 是免费提供的要求, 作者 ilaria.colombi@iit.it ilaria. colombi@iit。

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	<li>Fisher, R. 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=Operational+classification+of+seizure+types+by+the+International+League+Against+Epilepsy:+Position+Paper+of+the+ILAE+Commission+for+Classification+and+Terminology.">Operational classification of seizure types by the International League Against Epilepsy: Position Paper of the ILAE Commission for Classification and Terminology.</a> Epilepsia. 58 (4), 522-530 (2017).</li><li>WHO. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=.">.</a> Epilepsy Facts Sheet. , (2017).</li><li>Fiest, K. M., Birbeck, G. L., Jacoby, A., Jette, N. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Stigma+in+epilepsy.">Stigma in epilepsy.</a> Curr Neurol Neurosci Rep. 14 (5), 444 (2014).</li><li>Brodie, M. J., Barry, S. J., Bamagous, G. A., Norrie, J. D., Kwan, P. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Patterns+of+treatment+response+in+newly+diagnosed+epilepsy.">Patterns of treatment response in newly diagnosed epilepsy.</a> Neurology. 78 (20), 1548-1554 (2012).</li><li>Tanriverdi, T., Poulin, N., Olivier, A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Life+12+years+after+temporal+lobe+epilepsy+surgery:+a+long-term,+prospective+clinical+study.">Life 12 years after temporal lobe epilepsy surgery: a long-term, prospective clinical study.</a> Seizure. 17 (4), 339-349 (2008).</li><li>Dingledine, R. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=.">.</a> Brain slices. , (1984).</li><li>Avoli, 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=Network+and+pharmacological+mechanisms+leading+to+epileptiform+synchronization+in+the+limbic+system+in+vitro.">Network and pharmacological mechanisms leading to epileptiform synchronization in the limbic system in vitro.</a> Prog Neurobiol. 68 (3), 167-207 (2002).</li><li>Rutecki, P. A., Lebeda, F. J., Johnston, D. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Epileptiform+activity+induced+by+changes+in+extracellular+potassium+in+hippocampus.">Epileptiform activity induced by changes in extracellular potassium in hippocampus.</a> J Neurophysiol. 54 (5), 1363-1374 (1985).</li><li>Mody, I., Lambert, J. D., Heinemann, U. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Low+extracellular+magnesium+induces+epileptiform+activity+and+spreading+depression+in+rat+hippocampal+slices.">Low extracellular magnesium induces epileptiform activity and spreading depression in rat hippocampal slices.</a> J Neurophysiol. 57 (3), 869-888 (1987).</li><li>Huberfeld, G., 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=Glutamatergic+pre-ictal+discharges+emerge+at+the+transition+to+seizure+in+human+epilepsy.">Glutamatergic pre-ictal discharges emerge at the transition to seizure in human epilepsy.</a> Nat Neurosci. 14 (5), 627-634 (2011).</li><li>Pitkanen, A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=.">.</a> Models of seizures and epilepsy. , (2017).</li><li>Rutecki, P. A., Lebeda, F. J., Johnston, D. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=4-Aminopyridine+produces+epileptiform+activity+in+hippocampus+and+enhances+synaptic+excitation+and+inhibition.">4-Aminopyridine produces epileptiform activity in hippocampus and enhances synaptic excitation and inhibition.</a> J Neurophysiol. 57 (6), 1911-1924 (1987).</li><li>Gross, G. W., Rieske, E., Kreutzberg, G. W., Meyer, A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=A+new+fixed-array+multi-microelectrode+system+designed+for+long-term+monitoring+of+extracellular+single+unit+neuronal+activity+in+vitro.">A new fixed-array multi-microelectrode system designed for long-term monitoring of extracellular single unit neuronal activity in vitro.</a> Neurosci Lett. 6 (2-3), 101-105 (1977).</li><li>Pine, J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Recording+action+potentials+from+cultured+neurons+with+extracellular+microcircuit+electrodes.">Recording action potentials from cultured neurons with extracellular microcircuit electrodes.</a> J Neurosci Methods. 2 (1), 19-31 (1980).</li><li>Reinartz, S., Biro, I., Gal, A., Giugliano, M., Marom, S. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Synaptic+dynamics+contribute+to+long-term+single+neuron+response+fluctuations.">Synaptic dynamics contribute to long-term single neuron response fluctuations.</a> Front Neural Circuits. 8, 71 (2014).</li><li>Dossi, E., Blauwblomme, T., Nabbout, R., Huberfeld, G., Rouach, N. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Multi-electrode+array+recordings+of+human+epileptic+postoperative+cortical+tissue.">Multi-electrode array recordings of human epileptic postoperative cortical tissue.</a> J Vis Exp. (92), (2014).</li><li>Boido, 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=Cortico-hippocampal+hyperexcitability+in+synapsin+I/II/III+knockout+mice:+age-dependency+and+response+to+the+antiepileptic+drug+levetiracetam.">Cortico-hippocampal hyperexcitability in synapsin I/II/III knockout mice: age-dependency and response to the antiepileptic drug levetiracetam.</a> Neuroscience. 171 (1), 268-283 (2010).</li><li>Gonzalez-Sulser, 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=The+4-aminopyridine+in+vitro+epilepsy+model+analyzed+with+a+perforated+multi-electrode+array.">The 4-aminopyridine in vitro epilepsy model analyzed with a perforated multi-electrode array.</a> Neuropharmacology. 60 (7-8), 1142-1153 (2011).</li><li>Hajos, N., 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=Maintaining+network+activity+in+submerged+hippocampal+slices:+importance+of+oxygen+supply.">Maintaining network activity in submerged hippocampal slices: importance of oxygen supply.</a> Eur J Neurosci. 29 (2), 319-327 (2009).</li><li>Ivanov, A., Zilberter, Y. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Critical+state+of+energy+metabolism+in+brain+slices:+the+principal+role+of+oxygen+delivery+and+energy+substrates+in+shaping+neuronal+activity.">Critical state of energy metabolism in brain slices: the principal role of oxygen delivery and energy substrates in shaping neuronal activity.</a> Front Neuroenergetics. 3, 9 (2011).</li><li>MultichannelSystems. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=.">.</a> Manual PH01. , (2018).</li><li>Zhang, 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=Propagation+of+epileptiform+activity+can+be+independent+of+synaptic+transmission,+gap+junctions,+or+diffusion+and+is+consistent+with+electrical+field+transmission.">Propagation of epileptiform activity can be independent of synaptic transmission, gap junctions, or diffusion and is consistent with electrical field transmission.</a> J Neurosci. 34 (4), 1409-1419 (2014).</li><li>Panuccio, G., 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=In+vitro+ictogenesis+and+parahippocampal+networks+in+a+rodent+model+of+temporal+lobe+epilepsy.">In vitro ictogenesis and parahippocampal networks in a rodent model of temporal lobe epilepsy.</a> Neurobiol Dis. 39 (3), 372-380 (2010).</li><li>Barbarosie, M., Avoli, M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=CA3-driven+hippocampal-entorhinal+loop+controls+rather+than+sustains+in+vitro+limbic+seizures.">CA3-driven hippocampal-entorhinal loop controls rather than sustains in vitro limbic seizures.</a> J Neurosci. 17 (23), 9308-9314 (1997).</li><li>Panuccio, G., Guez, A., Vincent, R., Avoli, M., Pineau, J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Adaptive+control+of+epileptiform+excitability+in+an+in+vitro+model+of+limbic+seizures.">Adaptive control of epileptiform excitability in an in vitro model of limbic seizures.</a> Exp Neurol. 241, 179-183 (2013).</li><li>Benini, R., Avoli, M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Rat+subicular+networks+gate+hippocampal+output+activity+in+an+in+vitro+model+of+limbic+seizures.">Rat subicular networks gate hippocampal output activity in an in vitro model of limbic seizures.</a> J Physiol. 566 (Pt 3), 885-900 (2005).</li><li>D'Arcangelo, G., Panuccio, G., Tancredi, V., Avoli, M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Repetitive+low-frequency+stimulation+reduces+epileptiform+synchronization+in+limbic+neuronal+networks.">Repetitive low-frequency stimulation reduces epileptiform synchronization in limbic neuronal networks.</a> Neurobiol Dis. 19 (1-2), 119-128 (2005).</li><li>Steidl, E. M., Neveu, E., Bertrand, D., Buisson, B. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=The+adult+rat+hippocampal+slice+revisited+with+multi-electrode+arrays.">The adult rat hippocampal slice revisited with multi-electrode arrays.</a> Brain Res. 1096 (1), 70-84 (2006).</li><li>Pulizzi, R., 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=Brief+wide-field+photostimuli+evoke+and+modulate+oscillatory+reverberating+activity+in+cortical+networks.">Brief wide-field photostimuli evoke and modulate oscillatory reverberating activity in cortical networks.</a> Sci Rep. 6, 24701 (2016).</li><li>Liu, M. G., Chen, X. F., He, T., Li, Z., Chen, J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Use+of+multi-electrode+array+recordings+in+studies+of+network+synaptic+plasticity+in+both+time+and+space.">Use of multi-electrode array recordings in studies of network synaptic plasticity in both time and space.</a> Neuroscience Bulletin. 28 (4), 409-422 (2012).</li><li>Maier, N., Morris, G., Johenning, F. W., Schmitz, D. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=An+approach+for+reliably+investigating+hippocampal+sharp+wave-ripples+in+vitro.">An approach for reliably investigating hippocampal sharp wave-ripples in vitro.</a> PLoS One. 4 (9), e6925 (2009).</li><li>Aiba, I., Noebels, J. L. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Spreading+depolarization+in+the+brainstem+mediates+sudden+cardiorespiratory+arrest+in+mouse+SUDEP+models.">Spreading depolarization in the brainstem mediates sudden cardiorespiratory arrest in mouse SUDEP models.</a> Sci Transl Med. 7 (282), 282ra246 (2015).</li><li>MA, R., Avoli, M., Noebels, J. L., Rogawski, M. A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=.">.</a> Jasper's Basic Mechanisms of the Epilepsies . , (2012).</li><li>Motamedi, G. 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=Termination+of+epileptiform+activity+by+cooling+in+rat+hippocampal+slice+epilepsy+models.">Termination of epileptiform activity by cooling in rat hippocampal slice epilepsy models.</a> Epilepsy Res. 70 (2-3), 200-210 (2006).</li><li>Yang, X. F., Duffy, D. W., Morley, R. E., Rothman, S. M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Neocortical+seizure+termination+by+focal+cooling:+temperature+dependence+and+automated+seizure+detection.">Neocortical seizure termination by focal cooling: temperature dependence and automated seizure detection.</a> Epilepsia. 43 (3), 240-245 (2002).</li></ol>

作者没有什么可透露的。

多边环境协定是神经生理学调查的宝贵工具, 近年来由于勘探和发展已达到成熟。与常规场电位记录相比, 多边环境协定提供了较高的观测点和已知电极间距离的优势, 这对于准确定位神经元网络的相互作用至关重要。
该技术还可以结合其他电生理学方法, 如膜片钳记录15, 以研究单神经元和神经网络活动之间的关系。同时, 可视化场电位和多单位活动的可能性可以为小神经元群的活动与集体神经网络的相关性提供宝贵的见解。与电压敏感染料、钙成像和光遗传学29的结合允许使用多面体方法推断神经生理学现象.除了药理学研究, 在同一系统中进行记录和刺激的可能性使得该技术的记录技巧非常强大和多才多艺: 例如, 有可能研究突触可塑性现象在内存形成的核心30, 使用此处介绍的调节协议, 与 DBS 治疗癫痫和多种脑部疾病有关。最近的证据表明, 多边环境协定记录技术可以成功地应用于人类癫痫脑组织16表明它在癫痫研究中的宝贵用处, 既了解这种破坏性疾病的基本机制并微调 DBS 算法来改善它。
然而, 由于水下记录室的要求, 以及让大脑切片在微电极的固体基底上休息的必要性, 多边环境协定迫使对脑切片的 "极限" 条件下的电生理学实验进行追求。是集成的。因此, 脑组织可能无法获得足够的氧气供应, 这反过来又可能影响录音的质量。
这里描述的协议允许可靠地记录和研究 ictogenesis 在鼠边缘网络上的耦合, 利用急性体外4AP 模型和追求长期的电刺激, 为评估提供相关信息星展的政策。
以前对癫痫边缘网络的电调节的研究基于传统的场电位记录, 使用了老鼠或鼠脑切片, 结果类似24,25;但是, 由于与小鼠7获得的脑组织相比, 大鼠脑组织的使用被证明更具挑战性。关于多边环境协定技术, 小鼠脑切片最适合的优点是它们的体积较小。此外, 鉴于小鼠与大鼠脑组织中发作放电的发生率较高, 在一个实验日进行大量的脑切片测试是可能的, 这反过来又使数据收集速度更快、效率更高, 并能减少使用的动物数量。
对于现有方法的意义:
使用此处描述的自定义分庭记录的发作排放似乎与使用传统的多边环境协定环室和相同的多边环境协定类型 (平面微电极cf 17) 所观察到的持续时间和发生率相似。然而, 需要强调的是, 当使用常规的圆形记录室和低灌注率 (1 毫升/分钟) 时, 脑切片厚度必须显著降低, 这可能会阻碍调节实验的成功追求, 因为连接不良。
在本协议中, 由膜片钳记录室设计启发的低容量自定义记录室提供了稳定和可靠的层流, 这对于成功地追求多边环境协定录音至关重要;它还允许增加脑切片厚度为400µm, 以实现公平权衡组织的生存能力和内在的连通性。确实认识到, 腔内记录解决方案的层流是非常可取的脑切片电生理学, 因为它不受温度, 氧气和 pH 梯度的影响, 在循环流圆记录观察钱伯斯19,20,31 (像提供了商业上可用的多边环境协定)。这种梯度引入实验性偏倚, 也对脑切片有害。相对较小体积 (~ 1.5 毫升) 的记录室连同高灌注率 (5-6 毫升/分)16,31允许充分交换灌注介质 (≥3时间/分钟)。定制室可以很容易地获得从商业来源以实惠的价格或生产内部使用3D 印刷技术。其他研究报告了从400µm 厚的人脑切片16使用传统的多边环境协定环室的录音, 同时保持 ACSF 体积为1毫升, 并在低噪声蠕动泵的帮助下执行高灌注率 (5-6 毫升/分钟)。但是, 作者使用了不同的 ictogenesis 模型即, 低镁2 +, 这可能比4AP 模型的影响较小, ephaptic 机制涉及显著增加细胞外 K+浓度11,22. 我们发现, 4AP 模型中不需要高的灌注率, 可能是由于累积的胞外 K+的快速冲洗, 在录制 ACSF16中可能要显著增加。事实上, 当 ASCF 灌注速度增加到 2-3 毫升/分钟, 而脑切片被厚 ACSF 层淹没时, 发作事件出现了更多的 "块", 而在返回到较低的灌注率 (1 毫升/分钟) 和较薄的 ACSF 层在组织表面水平 (没有显示数据)。本协议中描述的自定义记录室允许以1毫升/分钟的流速交换灌注介质 3-5 次/分钟。因此, 即使在相对较低的灌注率下, 脑切片的整体氧气供应也得到了强烈的改善, 同时仍然保证了记录温度和高信噪比的稳定。最重要的是, 可以将胞外 K+浓度保持为生理值。
ictogenesis 的4AP 模型不需要对 ACSF 离子成分进行任何重大修改, 例如降低 Mg2 +或增加 K+, 并提供了保持兴奋和抑制传输完好的独特优势。12, 鉴于谷氨酸和 GABAergic 网络在癫痫发作同步中的关键作用 (cf 7), 这一方面与癫癎的研究高度相关。本协议中使用的 4 ap ACSF 包含生理 K+浓度 (3.25 毫米) 和略低的镁2 +浓度比持有 ACSF (1 毫米与1.3 毫米)。这种浓度仍然是在报告的镁2 +浓度在啮齿动物脑脊液中的生理价值, 并在许多实验室使用 (见例如17,18)。对于所述协议的目的, 我们发现, 这种轻微的减少是首选的援助, 以4AP 的效果。
在以前的工作中, 4AP 已被应用于脑切片, 当它已经定位在录音室17,18中。除非实验者需要观察4AP 应用与癫痫发作模式开始之间的时间滞后, 我们发现这种方法相当耗时, 不宜进行长时间的录音和刺激疗程。本议定书所述。32摄氏度的脑切片预孵化可节省大量实验时间, 因为脑切片可以在系列中预先处理, 同时使用其他组织切片进行实验。
脑切片的长期生存能力可能有助于分析长期的网络特征。此外, 如果实验者想要比较几种刺激方案, 这些方法必须在同一脑切片中进行, 以提高对重复电刺激的适应性, 这是很有好处的。在我们的手中, 当测试3刺激协议, 每一个之前的控制阶段, 然后是一个恢复阶段, 实验可能会持续 3-5 小时。在这种情况下, 实时制图是至关重要的, 以激活正确的途径和压制, 而不是青睐 ictogenesis 通过电刺激。我们的用户友好的 GUI 代表了一个快速, 简单和灵活的工具, 绘制不同的大脑结构的电极。与商业软件相反, 即使具有基本编程技能, 也可以添加自定义的多边环境协定布局。图像可以在明亮的领域获得;因此, 任何具有良好分辨率和适合显微镜的通用相机是合适的。倒置显微镜是必不可少的可视化的电极位置相对于脑切片, 因为这些将隐藏在组织下方, 如果使用直立显微镜。然而, 如果实验者需要进行刀切来扰乱特定的神经通路, 那么除了倒置型外, 还建议使用立体显微镜。
最后, 建议的方法的一个进一步好处是便宜和相对地容易装配多数必要的工具, 象自定义录音室和存放室、温暖的浴和切片锚, 以及不必要的使用昂贵的低噪音蠕动泵。
技术的局限性:
该记录不允许可视化非常慢的波浪,即, 信号中的 DC 移位。这种基线偏差可能有助于发作放电持续时间的渐进测量, 最重要的是, 它们是研究皮质扩张性抑郁症的基础 (一种与癫痫突发意外死亡有关的现象32 , 并共享癫痫与偏头痛之间33)。
关于电子调节, 这里所描述的协议允许在不影响脑切片生存能力的情况下进行几次刺激疗程。我们可以成功地执行多达3次的刺激会话, 20-45 分钟, 类似于以前工作中报告的25。虽然脑切片可能会经受更多的刺激疗程或更长的刺激协议, 但我们没有在这方面测试脑切片。我们建议将刺激方案的数量限制在 3, 避免延长 (≥60分钟) 刺激疗程, 这可能会显著地强调在这些限制条件下维持的脑组织, 但在刺激撤退时缺乏恢复。
议定书内的关键步骤:
几个关键因素可能会阻碍成功地追求记录和调制的癫痫活动的脑切片耦合到多边环境协定。除了所使用的啮齿动物种类和脑切片的质量, 录音过程中的灌注率和 ACSF 流的动态 (i. e, 层流与圆形) 在记录室内, 我们发现恢复的条件和脑切片的长期维护以及4AP 应用的模式是最关键的步骤。
在使用浸没式保温室时, 在室温下贮存脑切片, 以保持脑组织网络活动, 有利于4AP 应用发作放电的诱导, 是至关重要的。在我们的手, 恢复和维护在32°c 恶化了脑组织在 3-4 小时内切片。
然而, 4 ACSF 的脑切片在室温下孵化并记录在32摄氏度是不可取的。在这种情况下, 我们发现有许多困难在观察健壮的复发发作放电。事实上, 在20–24°c 范围内的低温已经报告, 以抑制或甚至防止 4 ap 诱发 ictogenesis 两个在体外34和在体内35。因此, 4AP 孵化和记录温度必须在 30-34 摄氏度以内, 必须匹配。为了避免把大脑组织置于太大的压力下, 由于同时诱导 hyperexcitability 4AP 和脑切片的突然暴露从室温到温暖 (32 °c) 4 ap-ACSF, 这是基本的执行中间预热步骤, 让大脑切片习惯在持有 ACSF 在32°c 20-30 分钟. 跳过此步骤可能会影响 ictogenesis 感应4AP。
另一个需要提及的方面是 ACSF 后的 d-葡萄糖浓度在切片后的重要性。25毫米的浓度保持脑切片优于10毫米浓度在许多实验室使用, 它也提高了发作活动的发生率, 这是2倍更快 (因此, 使其更容易进行长系列的实验规程在几脑切片每天)。
最后, 需要指出切片过程中的一些重要细节。首先, 不要让完整的大脑和脑切片冻结在冷切割 ACSF。为 < 2 分钟冷却的大脑是足够的, 因为小鼠的大脑大小。组织切片应立即从缓冲托盘转移到冲洗烧杯室温。冲洗切割 ACSF 是非常重要的, 否则高蔗糖浓度将使脑切片坚持在尼龙网眼的保持室。为了有效地冲洗组织, 重要的是尽量减少切割 ACSF 的内容内的转移吸管, 以避免污染的持有 ACSF 过度。2级冲洗有助于减少污染。切片程序完成后, 应立即将组织切片从冲洗烧杯转移到保温室, 在保持 ACSF 中, 软尼龙网暂停中途, 可使两侧的组织氧合。在切片过程后的所有阶段都应采用同样的原理: 脑切片不应该坐在烧杯的底部, 他们不应该接触到持有商会的两侧, 他们不应该接触对方, 也不应该重叠。
修改和疑难解答:
ACSF 成分可根据实验者的需要进行修改。例如, 可以添加药物来解剖特定的神经递质或离子通道对电调节的功效的贡献。此外, 可以省略丙酮和抗坏血酸 (cf 表 2), 尽管我们发现它们发挥了强大的神经保护作用。我们在表 3中报告了在本协议中可以遇到的最常见问题以及如何处理它们。
结论:
这一记录无疑是解决神经网络在健康和疾病中相互作用的宝贵技术。除了药理学研究, 还可以评估与星展集团应用于癫痫和其他神经系统疾病相关的电调节协议。在本议定书中, 我们已经表明, 有可能重现典型的周期性刺激实验, 其结果与常规细胞外磁场电位记录和外部刺激电极获得的效果相似。越来越多的商用用户友好设备和先进的软件工具, 使多边环境协定记录技术也适用于闭环刺激实验, 对脑组织提供特别刺激, 并研究反馈机制对神经网络反应的贡献。

癫痫是一种危及生命的渐进性紊乱, 导致大脑的不受控制的活动1;它携带着最高的疾病负担和重要的社会耻辱2,3。这是最常见的综合征 (40%) 和最频繁 (~ 30%) 抗癫痫药物治疗4。虽然癫痫组织的手术消融可以改善病人的病情, 但在所有患者中都是行不通的, 可能不能保证完全无癫痫的生活5。在药物治疗或神经外科不合适的情况下, 电星展集团对癫痫边缘网络的调制是一种很有前途的方法。
啮齿动物脑切片是研究神经元网络如何在健康和疾病中发挥作用的重要工具6通过体外电生理学技术, 因为它们保存, 至少部分, 大脑的原始结构和连通性感兴趣的区域 (ROI)。特别是, 水平结合的海马-嗅皮质 (海马-EC) 切片构成了所涉及的必要的神经网络, 因此在体外研究中经常使用7。
通过改变人工脑脊液 (ACSF) 的离子组成, 例如降低镁, 同时增加钾8,9,10或通过药理操作, 例如阻止抑制 GABAergic 活动 (参见11进行全面审查)。然而, 这些模型是基于激励和抑制的不平衡变化;因此, 他们不允许研究兴奋和抑制网络对 ictogenesis 的相互作用和协同作用。连续灌注脑切片与惊厥药物4AP 增强兴奋和抑制神经传递, 从而使研究急性 ictogenesis, 同时保持突触活动整体完整完好 12.
多边环境协定允许记录神经网络产生的电子活动从更多的观察点相比, 传统的细胞外场电位记录, 其中空间限制限制的电极数量, 可以在脑切片表面容纳。此外, 多边协定芯片提供了已知的电极间距离的额外优势, 这对于跟踪传播和评估记录信号的行进速度非常有用。虽然最初构思为记录从培养的神经元13,14, 多边环境协定现在也被用来表征从啮齿动物15和人类16获得的急性脑切片的电生理特征。因此, 在癫痫研究的背景下, 多边环境协定是一个有价值的工具, 用于查明神经网络在 ictogenesis16、17、18的核心中的相互作用。
然而, 在长时间 (几个小时) 实验协议中, 在获取或维持稳定的癫痫模式方面, 多边环境协定的记录本身就具有技术挑战。首先, 在大型和圆形浸没式记录室中, 组织氧合可能不足19,20典型的商用多边环境协定, 而低信噪比和温度不稳定可能影响记录的质量, 当一个高灌注率 (6-10 毫升/分钟) 是用来改善氧气供应的脑切片 (如请参见加热和灌注设备的技术说明21)。第二, 使用浸没式记录室19时很难观察到经常放电, 如癫痫放电等高频成分;当急性癫痫发作模式被化学诱导并涉及 ephaptic 机制时, 这种情况尤其如此, 因为4AP 模型22 (请参见11进行全面概述)。为了克服这些局限性, 研究人员提出了一些策略。例如, 增加灌注速率19、20 , 同时减少脑切片厚度 (≤300µM) 和区域17,18是实现组织充分氧气供应的关键因素。此外, 还可以通过使用穿孔的多边环境协定 (pMEAs) 来改善脑切片的生存能力, 从而使灌注的大脑组织从两侧18。
虽然所述方法大大提高了从脑切片记录的可行性, 但没有对长时间 (数小时) 记录和电刺激会话进行测试, 后者代表了一个重要大脑组织的压力。可能需要长时间的记录来研究癫痫模式的特定特征在时间上的演变, 这是不可能通过短期测量来揭露的。在星展研究的背景下, 可能需要长时间的实验协议来评估和比较几种刺激范式在同一脑切片中的作用。
当只有自发的电气活动需要记录时, 在数据分析过程中, 电极的映射通常是后验,即;相反, 对诱发反应或电调节范式的研究需要将刺激传递给特定的 ROI, 从而在实验过程中要求快速、容易的带电电极映射。
在这里, 我们说明了一个简单的实验协议, 允许诱导和维持一个稳定的 4 ap 诱发癫痫模式的成年啮齿动物脑切片。观察的活动忠实地再现了该模型的电生理特性, 其特点是使用常规的细胞外电生理技术。它持续了几个小时和经久持续的电刺激为重复的长的世纪。用于维持和孵化脑切片的分庭可以使用标准实验室用品 (图 1A) 轻松组装, 而一个允许最佳解决方案汇率和层流流的自定义记录室 (图 1B) 可以获得从商业来源或使用3D 印刷技术以实惠的价格。快速映射的 ROI (s) 的实时监测和选择的刺激电极是由一个自定义用户友好的 GUI 命名为mapMEA, 可以自由地根据请求提供。
<img alt="Figure 1" class="xfigimg" src="/files/ftp_upload/57548/57548fig1.jpg"> 图 1: 用于此协议的自定义设备.(A) 在4AP 中, 用于恢复、预热和预孵化的保温室采用烧杯和培养皿组装而成。培养皿的直径应小于断路器, 并通过注射器柱塞来保持到位。培养皿的底部被一个尼龙网 (从柔软的丝袜) 所取代, 用氰粘在培养皿的边缘上。氧气通过弯曲的脊椎针 (22 克) 提供, 插入在培养皿和烧杯的壁之间。气泡应该从烧杯的侧面冒出来, 永远不会到达尼龙网, 以避免大脑切片移位。培养皿的顶部可以作为盖子, 以避免 ACSF 蒸发和保持氧气饱和度。(B) 自定义记录室。在: 入口水库容纳热化套管和参考电极。出: 出水口水库, 可容纳吸针。录音室。( C) 玻璃巴斯德吸管, 由火抛光卷曲, 用于处理组织并调整其在录音室中的位置。( D) 自定义切片保留锚点。(E) 安装在多边环境协定芯片上的记录室的最终组装。脑切片靠在固定位置的底部。红色箭头表示覆盖加热套管的聚四氟乙烯管, 而红色圆圈表示参考电极, 这是 ACSF 在进气库中浸没的饱和氯化钾颗粒。蓝色箭头表示出入口储层中的吸力针。<a href="/files/ftp_upload/57548/57548fig1large.jpg" target="_blank">请单击此处查看此图的较大版本.</a>

<table><tbody><tr><td>Poly-D-Lysine&amp;nbsp;</td><td>Sigma-Adrich</td><td>P7886</td><td>Needed for MEA coating</td></tr><tr><td>NaCl</td><td>Sigma-Adrich</td><td>S9888</td><td>Chemical</td></tr><tr><td>KCl</td><td>Sigma-Adrich</td><td>P9541</td><td>Chemical</td></tr><tr><td>KH&lt;sub&gt;2&lt;/sub&gt;PO&lt;sub&gt;4&lt;/sub&gt;</td><td>Sigma-Adrich</td><td>795488</td><td>Chemical</td></tr><tr><td>CaCl&lt;sub&gt;2&lt;/sub&gt;*2 H&lt;sub&gt;2&lt;/sub&gt;O</td><td>Sigma-Adrich</td><td>C3306</td><td>Chemical</td></tr><tr><td>D-Glucose</td><td>Sigma-Adrich</td><td>RDD016</td><td>Chemical</td></tr><tr><td>NaHCO&lt;sub&gt;3&lt;/sub&gt;</td><td>Sigma-Adrich</td><td>S5761</td><td>Chemical</td></tr><tr><td>MgCl&lt;sub&gt;2&lt;/sub&gt;*6 H&lt;sub&gt;2&lt;/sub&gt;O</td><td>Sigma-Adrich</td><td>M2670</td><td>Chemical</td></tr><tr><td>MgSO&lt;sub&gt;4&lt;/sub&gt;*6 H&lt;sub&gt;2&lt;/sub&gt;O</td><td>Sigma-Adrich</td><td>M5921</td><td>Chemical</td></tr><tr><td>Sucrose</td><td>Sigma-Adrich</td><td>RDD023</td><td>Chemical</td></tr><tr><td>Pyruvic Acid, 98%&amp;nbsp;</td><td>Sigma-Adrich</td><td>107360</td><td>Chemical</td></tr><tr><td>4-aminopyridine</td><td>Sigma-Adrich</td><td>A78403</td><td>Convulsant drug</td></tr><tr><td>STG-2004</td><td>Multichannel System</td><td>STG4004-1.6mA</td><td>4-channel stimulus generator with voltage (&amp;plusmn;8 V) and current output (&amp;plusmn;1.6 mA)</td></tr><tr><td>MEA1060</td><td>Multichannel System</td><td>N/A</td><td>MEA amplifier</td></tr><tr><td>planar MEA</td><td>Multichannel System</td><td>60MEA500/30iR-Ti w/o ring</td><td>Must be without ring to allow using the custom recordingchamber</td></tr><tr><td>McRack</td><td>Multichannel System</td><td></td><td>Recording software</td></tr><tr><td>McStimulus II</td><td>Multichannel System</td><td></td><td>STG4004 control software</td></tr><tr><td>TC02</td><td>Multichannel System</td><td>TC02</td><td>2-channel thermostat</td></tr><tr><td>PH01</td><td>Multichannel System</td><td>PH01</td><td>Heating perfusion canula</td></tr><tr><td>MPH&amp;nbsp;</td><td>Multichannel System</td><td>MPH&amp;nbsp;</td><td>Magnetic holder for PH01 and suction needle</td></tr><tr><td>Elastostil E43</td><td>Wacker</td><td>E43</td><td>Elastomeric sealant used to mount the custom recording chamber onto the MEA</td></tr><tr><td>MEA Custom Chamber</td><td>Crisel Instrument</td><td>SKE-chamber MEA</td><td>Custom recording chamber</td></tr><tr><td>Ag/AgCl electrode, pellet, 1.0 mm&amp;nbsp;</td><td>Crisel Instrument</td><td>64-1309</td><td>Reference electrode for the custom recording chamber</td></tr><tr><td>Tergazyme</td><td>Sigma-Adrich</td><td>Z273287-1EA</td><td>Enzymatic cleaner</td></tr><tr><td>MATLAB</td><td>The Mathworks</td><td></td><td>Programming environment for electrodemapping</td></tr><tr><td>VT1000S</td><td>Leica Biosystems</td><td>VT1000S</td><td>Vibratome</td></tr><tr><td></td><td>Warner Instruments</td><td>64-1309</td><td>Ag-AgCl Electrode Pellet 1.0 mm (E205). Reference electrode.</td></tr></tbody></table>

recording and modulation of epileptiform activity in rodent brain slices coupled to microelectrode arrays

颞叶癫痫是最常见的部分复杂癫痫综合征, 对药物反应最少。深脑刺激 (DBS) 是一种有希望的方法, 当药理治疗失败或神经外科不建议。急性脑切片与微电极阵列 (多边环境协定) 相结合, 是研究神经网络相互作用及其电刺激调制的重要工具。与传统的细胞外记录技术相比, 它们提供了更多的观察点和已知的电极间距离的优势, 这使得研究的传播路径和电生理速度信号。然而, 在这一记录中, 组织氧合可能会受到很大的损害, 需要高的灌注率, 这是在实验温度降低信噪比和高振荡的代价。电刺激进一步强调脑组织, 使其难以追求长期记录/刺激时代。此外, 脑切片活动的电调制需要针对脑切片中的特定结构/通路, 要求电极映射在实验过程中能够轻松快速地进行。在这里, 我们说明如何使用平面多边环境协定对 4-氨基吡啶 (4 ap) 诱导的啮齿动物脑切片中的癫痫活动进行记录和电调制。我们表明, 从小鼠获得的脑组织优于大鼠脑组织, 因此更适合于实验。该协议保证了一种稳定的癫痫模式的产生和维持, 它能忠实地再现常规领域电位记录观察到的电生理特征, 持续数小时, 经久持续电刺激为长的世纪。由于使用小体积自定义记录室, 即使在低 (1 毫升/分钟) 灌注率下, 也可以实现整个实验的组织生存能力。实时监测和选择刺激电极的快速图象映射是由自定义图形用户界面 (GUI) 执行的。

Watch this Scientific Journal Video about Recording and Modulation of Epileptiform Activity in Rodent Brain Slices Coupled to Microelectrode Arrays at JoVE.com

Recording and Modulation of Epileptiform Activity in Rodent Brain Slices Coupled to Microelectrode Arrays

我们说明如何使用微电极阵列对 4-氨基吡啶诱导的啮齿动物脑切片中的癫痫活性进行记录和电调制。一个自定义记录室保持组织的可行性在长期的实验课。实时电极映射和激发对的选择由自定义图形用户界面执行。

鼠脑切片偶联微电极阵列癫痫活性的记录与调控

Temporal lobe epilepsy (TLE) is the most common partial complex epileptic syndrome and the least responsive to medications. Deep brain stimulation (DBS) ...

recording-modulation-epileptiform-activity-rodent-brain-slices

Research

JoVE Journal

Neuroscience

14.5K 次观看.  Istituto Italiano di Tecnologia. 我们说明如何使用微电极阵列对 4-氨基吡啶诱导的啮齿动物脑切片中的癫痫活性进行记录和电调制。一个自定义记录室保持组织的可行性在长期的实验课。实时电极映射和激发对的选择由自定义图形用户界面执行。

视频: Recording and Modulation of Epileptiform Activity in Rodent Brain Slices Coupled to Microelectrode Arrays

Multi-electrode Array Recordings of Human Epileptic Postoperative Cortical Tissue

Epilepsy, affecting about 1% of the population, comprises a group of neurological disorders characterized by the periodic occurrence of seizures, which disrupt normal brain function. Despite treatment with currently available antiepileptic drugs targeting neuronal functions, one third of patients with epilepsy are pharmacoresistant. In this condition, surgical resection of the brain area generating seizures remains the only alternative treatment. Studying human epileptic tissues has contributed to understand new epileptogenic mechanisms during the last 10 years. Indeed, these tissues generate spontaneous interictal epileptic discharges as well as pharmacologically-induced ictal events which can be recorded with classical electrophysiology techniques. Remarkably, multi-electrode arrays (MEAs), which are microfabricated devices embedding an array of spatially arranged microelectrodes, provide the unique opportunity to simultaneously stimulate and record field potentials, as well as action potentials of multiple neurons from different areas of the tissue. Thus MEAs recordings offer an excellent approach to study the spatio-temporal patterns of spontaneous interictal and evoked seizure-like events and the mechanisms underlying seizure onset and propagation. Here we describe how to prepare human cortical slices from surgically resected tissue and to record with MEAs interictal and ictal-like events ex vivo.

We here describe how to perform multi-electrode array recordings of human epileptic cortical tissue. Epileptic tissue resection, slice preparation and multi-electrode array recordings of interictal and ictal events are demonstrated in detail.

人类癫痫术后皮层组织的多电极阵列录音

Epilepsy, affecting about 1% of the population, comprises a group of neurological disorders characterized by the periodic occurrence of seizures, which ...

科研

医学

Direct-current Stimulation and Multi-electrode Array Recording of Seizure-like Activity in Mice Brain Slice Preparation

Cathodal transcranial direct-current stimulation (tDCS) induces suppressive effects on drug-resistant seizures. To perform effective actions, the stimulation parameters (e.g., orientation, field strength, and stimulation duration) need to be examined in mice brain slice preparations. Testing and arranging the orientation of the electrode relative to the position of the mice brain slice are feasible. The present method preserves the thalamocingulate pathway to evaluate the effect of DCS on anterior cingulate cortex seizure-like activities. The results of the multichannel array recordings indicated that cathodal DCS significantly decreased the amplitude of the stimulation-evoked responses and duration of 4-aminopyridine and bicuculline-induced seizure-like activity. This study also found that cathodal DCS applications at 15 min caused long-term depression in the thalamocingulate pathway. The present study investigates the effects of DCS on thalamocingulate synaptic plasticity and acute seizure-like activities. The current procedure can test the optimal stimulation parameters including orientation, field strength, and stimulation duration in an in vitro mouse model. Also, the method can evaluate the effects of DCS on cortical seizure-like activities at both the cellular and network levels.

Studies have shown that cathodal transcranial direct-current stimulation can produce suppressive effects on drug-resistant seizures. In this study, an in vitro experimental setup was devised in which the direct-current stimulation and multielectrode array recording of seizure-like activity were evaluated in mice brain slice preparation. The direct-current stimulation parameters were evaluated.

小鼠脑切片准备扣押样活性的直接电流刺激和多电极阵列记录

Cathodal transcranial direct-current stimulation (tDCS) induces suppressive effects on drug-resistant seizures. To perform effective actions, the ...

神经科学

Recording Network Activity in Spinal Nociceptive Circuits Using Microelectrode Arrays

The roles and connectivity of specific types of neurons within the spinal cord dorsal horn (DH) are being delineated at a rapid rate to provide an increasingly detailed view of the circuits underpinning spinal pain processing. However, the effects of these connections for broader network activity in the DH remain less well understood because most studies focus on the activity of single neurons and small microcircuits. Alternatively, the use of microelectrode arrays (MEAs), which can monitor electrical activity across many cells, provides high spatial and temporal resolution of neural activity. Here, the use of MEAs with mouse spinal cord slices to study DH activity induced by chemically stimulating DH circuits with 4-aminopyridine (4-AP) is described. The resulting rhythmic activity is restricted to the superficial DH, stable over time, blocked by tetrodotoxin, and can be investigated in different slice orientations. Together, this preparation provides a platform to investigate DH circuit activity in tissue from na&#239;ve animals, animal models of chronic pain, and mice with genetically altered nociceptive function. Furthermore, MEA recordings in 4-AP-stimulated spinal cord slices can be used as a rapid screening tool to assess the capacity of novel antinociceptive compounds to disrupt activity in the spinal cord DH.

The combined use of microelectrode array technology and 4-aminopyridine-induced chemical stimulation for investigating network-level nociceptive activity in the spinal cord dorsal horn is outlined.

使用微电极阵列记录脊髓伤害感受回路中的网络活动

The roles and connectivity of specific types of neurons within the spinal cord dorsal horn (DH) are being delineated at a rapid rate to provide an ...

小鼠运动皮层中的细胞外电生理记录

Multichannel Extracellular Recording in Freely Moving Mice

The protocol aims to uncover the properties of neuronal firing and network local field potentials (LFPs) in behaving mice carrying out specific tasks by correlating the electrophysiological signals with spontaneous and/or specific behavior. This technique represents a valuable tool in studying the neuronal network activity underlying these behaviors. The article provides a detailed and complete procedure for electrode implantation and consequent extracellular recording in free-moving conscious mice. The study includes a detailed method for implanting the microelectrode arrays, capturing the LFP and neuronal spiking signals in the motor cortex (MC) using a multichannel system, and the subsequent offline data analysis. The advantage of multichannel recording in conscious animals is that a greater number of spiking neurons and neuronal subtypes can be obtained and compared, which allows the evaluation of the relationship between a specific behavior and the associated electrophysiological signals. Notably, the multichannel extracellular recording technique and the data analysis procedure described in the present study can be applied to other brain areas when conducting experiments in behaving mice.

作者聚焦:用于研究自由移动小鼠神经元活动的多通道细胞外记录的进展 

The protocol describes the methodology of extracellular recording in the motor cortex (MC) to reveal extracellular electrophysiological properties in freely moving conscious mice, as well as the data analysis of local field potentials (LFPs) and spikes, which is useful for evaluating the network neural activity underlying behaviors of interest.

自由移动小鼠的多通道细胞外记录

The protocol aims to uncover the properties of neuronal firing and network local field potentials (LFPs) in behaving mice carrying out specific tasks by ...

<meta charset="utf8"></head><body>记录大脑组件中的网络活动：多电极阵列方法

A Multi-Electrode Array Platform for Modeling Epilepsy Using Human Pluripotent Stem Cell-Derived Brain Assembloids

Human brain organoids are three-dimensional (3D) structures derived from human pluripotent stem cells (hPSCs) that recapitulate&#160;aspects of fetal brain development. The fusion of dorsal with ventral regionally specified brain organoids in vitro generates assembloids, which have functionally integrated microcircuits with excitatory and inhibitory neurons. Due to their structural complexity and diverse population of neurons, assembloids have become a useful in vitro tool for studying aberrant network activity. Multi-electrode array (MEA) recordings serve as a method for capturing electrical field potentials, spikes, and longitudinal network dynamics from a population of neurons without compromising cell membrane integrity. However, adhering assembloids onto the electrodes for long-term recordings can be challenging due to their large size and limited contact surface area with the electrodes. Here, we demonstrate a method to plate assembloids onto MEA plates for recording electrophysiological activity over a 2-month span. Although the current protocol utilizes human cortical organoids, it can be broadly adapted to organoids differentiated to model other brain regions. This protocol establishes a robust, longitudinal, electrophysiological assay for studying the development of a neuronal network, and this platform has the potential to be used in drug screening for therapeutic development in epilepsy.

<meta charset="utf8"></head><body>作者聚焦：通过基于多电极阵列的人脑组件长期电生理监测推进遗传性癫痫研究

This protocol aims to stably plate dorsal-ventral fused assembloids on multi-electrode arrays for modeling epilepsy in vitro.

使用人类多能干细胞来源的脑组件对癫痫进行建模的多电极阵列平台

Human brain organoids are three-dimensional (3D) structures derived from human pluripotent stem cells (hPSCs) that recapitulate&#160;aspects of fetal ...

<meta charset="utf8"></head><body>使用 CMOS-HD-MEA 一致地诱导和记录癫痫样活动

High-Quality Seizure-Like Activity from Acute Brain Slices Using a Complementary Metal-Oxide-Semiconductor High-Density Microelectrode Array System

Complementary metal-oxide-semiconductor high-density microelectrode array (CMOS-HD-MEA) systems can record neurophysiological activity from cell cultures and ex vivo brain slices in unprecedented electrophysiological detail. CMOS-HD-MEAs were first optimized to record high-quality neuronal unit activity from cell cultures but have also been shown to produce quality data from acute retinal and cerebellar slices. Researchers have recently used CMOS-HD-MEAs to record local field potentials (LFPs) from acute, cortical rodent brain slices. One LFP of interest is seizure-like activity. While many users have produced brief, spontaneous epileptiform discharges using CMOS-HD-MEAs, few users reliably produce quality seizure-like activity. Many factors may contribute to this difficulty, including electrical noise, the inconsistent nature of producing seizure-like activity when using submerged recording chambers, and the limitation that 2D CMOS-MEA chips only record from the surface of the brain slice. The techniques detailed in this protocol should enable users to consistently induce and record high-quality seizure-like activity from acute brain slices with a CMOS-HD-MEA system. In addition, this protocol outlines the proper treatment of CMOS-HD-MEA chips, the management of solutions and brain slices during experimentation, and equipment maintenance.

<meta charset="utf8"></head><body>作者聚焦：使用 CMOS 高密度微电极阵列系统解开癫痫持续状态的癫痫发作动力学和新疗法

Here, we outline a protocol for using complementary metal-oxide-semiconductor high-density microelectrode array systems (CMOS-HD-MEAs)&#160;to record seizure-like activity from ex vivo brain slices.

使用互补金属氧化物半导体高密度微电极阵列系统从急性脑切片中获得高质量的癫痫样活性

Complementary metal-oxide-semiconductor high-density microelectrode array (CMOS-HD-MEA) systems can record neurophysiological activity from cell cultures ...

鼠脑切片偶联微电极阵列癫痫活性的记录与调控

摘要

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此视频中的章节

人类癫痫术后皮层组织的多电极阵列录音

小鼠脑切片准备扣押样活性的直接电流刺激和多电极阵列记录

使用微电极阵列记录脊髓伤害感受回路中的网络活动

自由移动小鼠的多通道细胞外记录

自由移动小鼠的多通道细胞外记录

自由移动小鼠的多通道细胞外记录

使用人类多能干细胞来源的脑组件对癫痫进行建模的多电极阵列平台

使用人类多能干细胞来源的脑组件对癫痫进行建模的多电极阵列平台

使用人类多能干细胞来源的脑组件对癫痫进行建模的多电极阵列平台

使用互补金属氧化物半导体高密度微电极阵列系统从急性脑切片中获得高质量的癫痫样活性