van Rensburg R, Errington D R, Ennaceur A, Lees G, Obrenovitch T P, Chazot P L
Centre for Integrative Neuroscience (CINS), School of Biological and Biomedical Sciences, Science Park, South Road, Durham University, Durham, County Durham DH1 3LE, United Kingdom.
J Neurosci Methods. 2009 Mar 15;177(2):311-6. doi: 10.1016/j.jneumeth.2008.10.012. Epub 2008 Oct 21.
Spreading depression (SD), whether elicited by local application of high K(+) medium to the cortical surface or by other stimuli, can increase the brain's tolerance to a subsequent, severe ischaemic insult in vivo, a phenomenon termed preconditioning. Herein, we have developed and validated a robust in vitro protocol for high-K(+)-preconditioning of cultured neurones. This new model is especially appropriate to unravel the molecular mechanisms underlying neuronal preconditioning and subsequent ischaemic tolerance. With this new, optimised preparation, preconditioning was found to be dependent upon culture day in vitro, cell density, K(+) concentration and duration of treatment. Finally, preconditioning was shown to be dependent upon N-methyl-d-aspartate (NMDA), CAM-kinase II signalling and alpha7-nicotinic (alpha7 nACh) receptor function, which is analogous to in vivo preconditioning induced by various stimuli.
无论是通过向皮质表面局部应用高钾培养基还是通过其他刺激引发的扩散性抑制（SD），均可增强大脑在体内对随后严重缺血性损伤的耐受性，这一现象称为预处理。在此，我们开发并验证了一种用于培养神经元高钾预处理的可靠体外实验方案。这种新模型特别适合于揭示神经元预处理及随后的缺血耐受性背后的分子机制。通过这种新的优化制剂，发现预处理取决于体外培养天数、细胞密度、钾离子浓度和处理持续时间。最后，结果表明预处理依赖于N-甲基-D-天冬氨酸（NMDA）、钙调蛋白依赖性蛋白激酶II信号传导和α7-烟碱（α7 nACh）受体功能，这类似于各种刺激在体内诱导的预处理。
