Jones Roland S G, da Silva Anderson Brito, Whittaker Roger G, Woodhall Gavin L, Cunningham Mark O
Department of Pharmacy and Pharmacology, University of Bath, Claverton Down, Bath BA2 7AY, UK.
Institute of Neuroscience, The Medical School, Framlington Place, Newcastle University, Newcastle upon Tyne, UK.
J Neurosci Methods. 2016 Feb 15;260:221-32. doi: 10.1016/j.jneumeth.2015.09.021. Epub 2015 Oct 3.
Increasingly, neuroscientists are taking the opportunity to use live human tissue obtained from elective neurosurgical procedures for electrophysiological studies in vitro. Access to this valuable resource permits unique studies into the network dynamics that contribute to the generation of pathological electrical activity in the human epileptic brain. Whilst this approach has provided insights into the mechanistic features of electrophysiological patterns associated with human epilepsy, it is not without technical and methodological challenges. This review outlines the main difficulties associated with working with epileptic human brain slices from the point of collection, through the stages of preparation, storage and recording. Moreover, it outlines the limitations, in terms of the nature of epileptic activity that can be observed in such tissue, in particular, the rarity of spontaneous ictal discharges, we discuss manipulations that can be utilised to induce such activity. In addition to discussing conventional electrophysiological techniques that are routinely employed in epileptic human brain slices, we review how imaging and multielectrode array recordings could provide novel insights into the network dynamics of human epileptogenesis. Acute studies in human brain slices are ultimately limited by the lifetime of the tissue so overcoming this issue provides increased opportunity for information gain. We review the literature with respect to organotypic culture techniques that may hold the key to prolonging the viability of this material. A combination of long-term culture techniques, viral transduction approaches and electrophysiology in human brain slices promotes the possibility of large scale monitoring and manipulation of neuronal activity in epileptic microcircuits.
越来越多的神经科学家开始利用从选择性神经外科手术中获取的人类活体组织进行体外电生理研究。获取这一宝贵资源有助于开展独特的研究，深入探究导致人类癫痫性大脑中病理性电活动产生的网络动力学。虽然这种方法为了解与人类癫痫相关的电生理模式的机制特征提供了见解，但它并非没有技术和方法上的挑战。这篇综述概述了从收集癫痫患者脑片开始，经过制备、储存和记录阶段，在使用癫痫患者脑片进行研究时所面临的主要困难。此外，它还概述了在此类组织中可观察到的癫痫活动性质方面的局限性，特别是自发性发作放电的罕见性，我们讨论了可用于诱发此类活动的操作方法。除了讨论在癫痫患者脑片中常规使用的传统电生理技术外，我们还回顾了成像和多电极阵列记录如何能够为人类癫痫发生的网络动力学提供新的见解。对人类脑片进行的急性研究最终受到组织寿命的限制，因此克服这一问题为获取更多信息提供了更多机会。我们回顾了关于器官型培养技术的文献，这些技术可能是延长这种材料活力的关键。长期培养技术、病毒转导方法和人类脑片电生理的结合，增加了对癫痫微回路中神经元活动进行大规模监测和操纵的可能性。
