Strauss Ulf, Kole Maarten H P, Bräuer Anja U, Pahnke Jens, Bajorat Rika, Rolfs Arndt, Nitsch Robert, Deisz Rudolf A
Department of Neurology, University Rostock, Gehlsheimer Strasse 20, 18147 Rostock, Germany.
Eur J Neurosci. 2004 Jun;19(11):3048-58. doi: 10.1111/j.0953-816X.2004.03392.x.
Neuronal subthreshold excitability and firing behaviour are markedly influenced by the activation and deactivation of the somato-dendritic hyperpolarization-activated cation current (Ih). Here, we evaluated possible contributions of Ih to hyperexcitability in an animal model of absence seizures (WAG/Rij rats). We investigated pyramidal neurons of the somatosensory neocortex, the site of generation of spike-wave discharges. Ih-mediated functions in neurons from WAG/Rij rats, Wistar rats (sharing the same genetic background with WAG/Rij, but less epilepsy-prone) and ACI rats (an inbred strain, virtually free of seizures) were compared. We complemented whole-cell recordings from layer 2-3 pyramidal neurons with immunohistochemistry, Western blot and RT-PCR analysis of the h-channel subunits HCN1-4. The fast component of Ih activation in WAG/Rij neurons was significantly reduced (50% reduction in the h-current density) and four times slower than in neurons from nonepileptic Wistar or ACI rats. The results showing decreases in currents corresponded to a 34% reduction in HCN1 protein in the WAG/Rij compared to the Wistar neocortex, but HCN1 mRNA showed stable expression. The other three Ih subunit mRNAs and proteins (HCN2-4) were not affected. The alterations in Ih magnitude and kinetics of gating in WAG/Rij neurons may contribute to augmented excitatory postsynaptic potentials, the increase in their temporal summation and the facilitation of burst firing of these neurons because each of these effects could be mimicked by the selective Ih antagonist ZD 7288. We suggest that the deficit in Ih-mediated functions may contribute to the development and onset of spontaneously occurring hyperexcitability in a rat model of absence seizures.
神经元阈下兴奋性和放电行为受到躯体 - 树突超极化激活阳离子电流(Ih)激活和失活的显著影响。在此,我们评估了Ih在失神癫痫动物模型(WAG/Rij大鼠)中对兴奋性过高的可能作用。我们研究了躯体感觉新皮层的锥体神经元,其是棘波放电产生的部位。比较了WAG/Rij大鼠、Wistar大鼠(与WAG/Rij具有相同遗传背景,但癫痫易感性较低)和ACI大鼠(近交系,几乎无癫痫发作)神经元中Ih介导的功能。我们用免疫组织化学、蛋白质印迹法以及对h通道亚基HCN1 - 4进行逆转录 - 聚合酶链反应(RT - PCR)分析,对第2 - 3层锥体神经元进行全细胞记录进行补充。WAG/Rij神经元中Ih激活的快速成分显著降低(h电流密度降低50%),且比非癫痫性Wistar或ACI大鼠的神经元慢四倍。电流降低的结果对应于与Wistar新皮层相比,WAG/Rij中HCN1蛋白减少34%,但HCN1 mRNA表达稳定。其他三种Ih亚基mRNA和蛋白质(HCN2 - 4)未受影响。WAG/Rij神经元中Ih大小和门控动力学的改变可能导致兴奋性突触后电位增强、其时间总和增加以及这些神经元爆发性放电的促进,因为这些效应均可被选择性Ih拮抗剂ZD 7288模拟。我们认为,Ih介导功能的缺陷可能导致失神癫痫大鼠模型中自发性兴奋性过高的发生和发作。