Department of Physiology and Pharmacology, Istituto Pasteur-Fondazione Cenci Bolognetti, University of Rome Sapienza, Rome, Italy.
Department of (Neuro)Pathology, Academic Medical Center, University of Amsterdam, The Netherlands.
Neurobiol Dis. 2016 Nov;95:93-101. doi: 10.1016/j.nbd.2016.07.014. Epub 2016 Jul 16.
Tuberous sclerosis complex (TSC) is a rare multi-system genetic disease characterized by several neurological disorders, the most common of which is the refractory epilepsy caused by highly epileptogenic cortical lesions. Previous studies suggest an alteration of GABAergic and glutamatergic transmission in TSC brain indicating an unbalance of excitation/inhibition that can explain, at least in part, the high incidence of epilepsy in these patients. Here we investigate whether TSC cortical tissues could retain GABAA and AMPA receptors at early stages of human brain development thus contributing to the generation and recurrence of seizures. Given the limited availability of pediatric human brain specimens, we used the microtransplantation method of injecting Xenopus oocytes with membranes from TSC cortical tubers and control brain tissues. Moreover, qPCR was performed to investigate the expression of GABAA and AMPA receptor subunits (GABAA α1-5, β3, γ2, δ; GluA1, GluA2) and cation chloride co-transporters NKCC1 and KCC2. The evaluation of nine human cortical brain samples, from 15 gestation weeks to 15years old, showed a progressive shift towards more hyperpolarized GABAA reversal potential (EGABA). This shift was associated with a differential expression of the chloride cotransporters NKCC1 and KCC2. Furthermore, the GluA1/GluA2 mRNA ratio of expression paralleled the development process. On the contrary, in oocytes micro-transplanted with epileptic TSC tuber tissue from seven patients, neither the GABAA reversal potential nor the GluA1/GluA2 expression showed similar developmental changes. Our data indicate for the first time, that in the same cohort of TSC patients, the pattern of both GABAAR and GluA1/GluA2 functions retains features that are typical of an immature brain. These observations support the potential contribution of altered receptor function to the epileptic disorder of TSC and may suggest novel therapeutic approaches. Furthermore, our findings strengthen the novel hypothesis that other developmental brain diseases can share the same hallmarks of immaturity leading to intractable seizures.
结节性硬化症(TSC)是一种罕见的多系统遗传性疾病,其特征为多种神经障碍,其中最常见的是由高度致痫性皮质病变引起的难治性癫痫。先前的研究表明,TSC 大脑中的 GABA 能和谷氨酸能传递发生改变,表明兴奋/抑制失衡,这至少可以部分解释这些患者癫痫的高发率。在这里,我们研究了 TSC 皮质组织是否可以在人类大脑发育的早期保留 GABAA 和 AMPA 受体,从而有助于癫痫发作的产生和复发。鉴于儿科人类大脑标本的有限可用性,我们使用微移植方法,将 TSC 皮质结节和对照脑组织的膜注入非洲爪蟾卵母细胞。此外,进行了 qPCR 以研究 GABAA 和 AMPA 受体亚基(GABAAα1-5、β3、γ2、δ;GluA1、GluA2)和阳离子氯离子共转运蛋白 NKCC1 和 KCC2 的表达。对来自 15 孕周至 15 岁的九个人类皮质脑样本的评估表明,GABAA 反转电位(EGABA)逐渐向更超极化方向转变。这种转变与氯离子共转运蛋白 NKCC1 和 KCC2 的差异表达有关。此外,GluA1/GluA2mRNA 的表达比例与发育过程平行。相反,在从七名患者的癫痫性 TSC 结节组织中微移植的卵母细胞中,GABAA 反转电位和 GluA1/GluA2 表达均未显示出类似的发育变化。我们的数据首次表明,在同一队列的 TSC 患者中,GABAA 受体和 GluA1/GluA2 功能的模式保留了典型的不成熟大脑的特征。这些观察结果支持改变的受体功能对 TSC 癫痫障碍的潜在贡献,并可能提示新的治疗方法。此外,我们的发现加强了新的假说,即其他发育性脑疾病可能具有相同的不成熟特征,导致难治性癫痫。