Institute of Neurophysiology, Charité Universitätsmedizin Berlin, Charitéplatz 1, D 10117 Berlin, Germany.
Neurobiol Dis. 2013 Aug;56:14-24. doi: 10.1016/j.nbd.2013.02.016. Epub 2013 Apr 11.
In models of temporal lobe epilepsy, in-vitro exposure of the entorhinal cortex (EC) to low concentrations of acetylcholine (ACh) induces muscarinic-dependent seizure-like events. Potassium channels from the KCNQ/Kv7 family, which close upon activation of muscarinic receptors, are mutated in several epileptic syndromes such as benign familial neonatal convulsions (KCNQ2/KCNQ3) and sudden unexplained death in epilepsy (KCNQ1). Therefore, we tested the hypothesis whether the ictogenic effect of ACh involves alterations of KCNQ channels. In horizontal temporo-hippocampal slices from pilocarpine-treated chronically epileptic rats, field potential recordings of epileptiform activity were performed in response to the application of ACh, the KCNQ blocker linopirdine, and KCNQ agonists. In the EC of control rats, ACh (20 and 50 μM) induced nested fast activity in the range of 15-20 Hz riding on <1 Hz slow oscillations. By contrast, in slices from pilocarpine-treated rats, 5 μM ACh was sufficient to induce interictal discharges that frequently transformed to epileptiform events at 20 μM ACh. While the non-specific KCNQ/Kv7 channel blocker linopirdine (20 and 50 μM) had no effect in control animals, in slices from epileptic rats it induced interictal discharges or seizure-like events. These could be blocked by the unspecific KCNQ/Kv7 agonist retigabine and attenuated by the Kv7.1 agonist L364-373. Immunohistochemistry revealed reduced expression of KCNQ2 and KCNQ3 in the EC and of KCNQ3-positive dendrites in the subiculum of epileptic rats. These results indicate that channels of the KCNQ family are key regulators of seizure susceptibility and their decreased availability in the epileptic tissue may reduce seizure threshold and contribute to ictogenesis.
在颞叶癫痫模型中,低浓度乙酰胆碱 (ACh) 体外暴露于内嗅皮层 (EC) 会诱导毒蕈碱依赖性癫痫样事件。在几种癫痫综合征中,如良性家族性新生儿惊厥 (KCNQ2/KCNQ3) 和癫痫伴突发性不明原因死亡 (KCNQ1),KCNQ/Kv7 家族的钾通道会发生突变,这些通道在毒蕈碱受体激活时关闭。因此,我们测试了 ACh 的致痫作用是否涉及 KCNQ 通道改变的假设。在从匹鲁卡品处理的慢性癫痫大鼠中获得的水平颞叶-海马切片中,对 ACh、KCNQ 阻断剂林匹啶和 KCNQ 激动剂的应用进行了癫痫样活动的场电位记录。在对照大鼠的 EC 中,ACh(20 和 50 μM)诱导了范围在 15-20 Hz 的嵌套快速活动,在<1 Hz 的慢振荡上骑行。相比之下,在来自匹鲁卡品处理大鼠的切片中,5 μM ACh 足以诱导发作间期放电,这些放电在 20 μM ACh 时经常转变为癫痫样事件。虽然非特异性 KCNQ/Kv7 通道阻断剂林匹啶(20 和 50 μM)在对照动物中没有作用,但在癫痫大鼠的切片中,它会引起发作间期放电或癫痫样事件。这些可以被非特异性 KCNQ/Kv7 激动剂 retigabine 阻断,并被 Kv7.1 激动剂 L364-373 减弱。免疫组织化学显示,在癫痫大鼠的 EC 中 KCNQ2 和 KCNQ3 的表达减少,在 subiculum 中 KCNQ3 阳性树突减少。这些结果表明,KCNQ 家族的通道是癫痫易感性的关键调节剂,它们在癫痫组织中的可用性降低可能会降低癫痫发作阈值并有助于癫痫发作。