de Curtis Marco, Librizzi Laura, Uva Laura
Unit of Epileptology and Experimental Neurophysiology, Fondazione Istituto Neurologico Carlo Besta, Milano, Italy.
Unit of Epileptology and Experimental Neurophysiology, Fondazione Istituto Neurologico Carlo Besta, Milano, Italy.
J Neurosci Methods. 2016 Feb 15;260:83-90. doi: 10.1016/j.jneumeth.2015.03.026. Epub 2015 Apr 2.
Research on ictogenesis is based on the study of activity between seizures and during seizures in animal models of epilepsy (chronic condition) or in in vitro slices obtained from naïve non-epileptic brains after treatment with pro-convulsive drugs, manipulations of the extracellular medium and specific stimulation protocols.
The in vitro isolated guinea pig brain retains the functional connectivity between brain structures and maintains interactions between neuronal, glial and vascular compartments. It is a close-to-in vivo preparation that offers experimental advantages not achieved with the use of other experimental models. Neurophysiological and imaging techniques can be utilized in this preparation to study brain activity during and between seizures induced by pharmacological or functional manipulations.
Cellular and network determinants of interictal and ictal discharges that reproduce abnormal patterns observed in human focal epilepsies and the associated changes in extracellular ion and blood-brain permeability can be identified and analyzed in the isolated guinea pig brain.
Ictal and interictal patterns recorded in in vitro slices may show substantial differences from seizure activity recorded in vivo due to slicing procedure itself. The isolated guinea pig brain maintained in vitro by arterial perfusion combines the typical facilitated access of in vitro preparations, that are difficult to approach during in vivo experiments, with the preservation of larger neuronal networks.
The in vitro whole isolated guinea pig brain preparation offers an unique experimental model to study systemic and neurovascular changes during ictogenesis.
癫痫发作起源的研究基于对癫痫动物模型(慢性疾病)发作期间及发作间隙的活动研究,或者基于对经惊厥药物处理、细胞外培养基操作及特定刺激方案处理后的未患癫痫的幼稚大脑获取的体外脑片的研究。
体外分离的豚鼠大脑保留了脑结构之间的功能连接,并维持神经元、神经胶质和血管成分之间的相互作用。这是一种接近体内状态的制备方法,具有使用其他实验模型无法实现的实验优势。可以利用神经生理学和成像技术在该制备方法中研究药理学或功能性操作诱导的癫痫发作期间及发作间隙的脑活动。
在分离的豚鼠大脑中,可以识别和分析发作间期和发作期放电的细胞和网络决定因素,这些因素再现了人类局灶性癫痫中观察到的异常模式以及细胞外离子和血脑通透性的相关变化。
由于切片过程本身,体外脑片中记录的发作期和发作间期模式可能与体内记录的癫痫发作活动有很大差异。通过动脉灌注维持在体外的分离豚鼠大脑,将体外制备方法典型的易于操作的特点(这在体内实验中很难实现)与更大神经元网络的保留相结合。
体外全分离豚鼠大脑制备方法提供了一个独特的实验模型,用于研究癫痫发作起源期间的全身和神经血管变化。