Department of Physiology, Feinberg School of Medicine, Northwestern University, Chicago, IL 60611, USA.
Cereb Cortex. 2021 Aug 26;31(10):4681-4698. doi: 10.1093/cercor/bhab115.
The decreased expression of the KCC2 membrane transporter in subicular neurons has been proposed to be a key epileptogenic event in temporal lobe epilepsy (TLE). Here, we have addressed this question in a reduced model in vitro and have studied the properties and mechanistic involvement of a major class of interneurons, that is, parvalbumin-expressing cells (PVs). When exposed to the KCC2 blocker VU0463271, mouse subicular slices generated hypersynchronous discharges that could be recorded electrophysiologically and visualized as clusters of co-active neurons with calcium imaging. The pharmacological profile of these events resembled interictal-like discharges in human epileptic tissue because of their dependence on GABAA and AMPA receptors. On average, PVs fired before pyramidal cells (PCs) and the area of co-active clusters was comparable to the individual axonal spread of PVs, suggesting their mechanistic involvement. Optogenetic experiments confirmed this hypothesis, as the flash-stimulation of PVs in the presence of VU0463271 initiated interictal-like discharges, whereas their optogenetic silencing suppressed network hyper-excitability. We conclude that reduced KCC2 activity in subicular networks in vitro is sufficient to induce interictal-like activity via altered GABAergic signaling from PVs without other epilepsy-related changes. This conclusion supports an epileptogenic role for impaired subicular KCC2 function during the progression of TLE.
在颞叶癫痫(TLE)中,突触旁神经元中 KCC2 膜转运体的表达减少被认为是致痫性事件的关键。在这里,我们在体外的简化模型中研究了这个问题,并研究了主要一类中间神经元,即表达钙调蛋白结合蛋白的细胞(PVs)的特性和机制参与。当暴露于 KCC2 阻断剂 VU0463271 时,小鼠海马切片产生超同步放电,这些放电可以用电生理学记录下来,并通过钙成像可视化成群的共同激活神经元。这些事件的药理学特征类似于人类癫痫组织中的发作间期放电,因为它们依赖于 GABA A 和 AMPA 受体。平均而言,PVs 在 PC 之前放电,并且共同激活簇的区域与 PVs 的单个轴突传播相当,这表明它们的机械参与。光遗传实验证实了这一假设,因为在 VU0463271 存在下,对 PVs 的闪光刺激引发了发作间期样放电,而它们的光遗传沉默抑制了网络超兴奋性。我们得出结论,体外海马网络中 KCC2 活性的降低足以通过改变来自 PVs 的 GABA 能信号来诱导发作间期样活性,而无需其他与癫痫相关的变化。这一结论支持在 TLE 进展过程中,受损的海马 KCC2 功能具有致痫作用。